You are on page 1of 70


Australian Journal of Botany, 2016, 64, 715–716

New handbook for standardised measurement of plant
functional traits worldwide

N. Pérez-Harguindeguy, S. Díaz, E. Garnier, S. Lavorel, H. Poorter, P. Jaureguiberry,
M. S. Bret-Harte, W. K. Cornwell, J. M. Craine, D. E. Gurvich, C. Urcelay, E. J. Veneklaas,
P. B. Reich, L. Poorter, I. J. Wright, P. Ray, L. Enrico, J. G. Pausas, A. C. de Vos, N. Buchmann,
G. Funes, F. Quétier, J. G. Hodgson, K. Thompson, H. D. Morgan, H. ter Steege,
M. G. A. van der Heijden, L. Sack, B. Blonder, P. Poschlod, M. V. Vaieretti, G. Conti,
A. C. Staver, S. Aquino and J. H. C. Cornelissen

(Vol. 61 no. 3 pp. 167–234)
The authors regret that elements of Appendix 1 were incorrect in the original publication. The correct version of Appendix 1 is
given below.

Appendix 1. Summary of plant traits
Summary of plant traits included in the handbook
The range of values corresponds to those generally reported for field-grown plants. Ranges of values are based on the literature and the authors’ datasets and do not
always necessarily correspond to the widest ranges that exist in nature or are theoretically possible. Recommended sample size indicates the minimum and
preferred number of individuals to be sampled, so as to obtain an appropriate indication of the values for the trait of interest; when only one value is given, it
corresponds to the number of individuals ( = replicates); when two values are given, the first one corresponds to the number of individuals and the second one to the
number of organs to be measured per individual. Note that one replicate can be compounded from several individuals (for smaller species), whereas one individual
cannot be used for different replicates. The expected coefficient of variation (CV) range gives the 20th and the 80th percentile of the CV ( = s.d. scaled to the mean)
as observed in several datasets obtained for a range of field plants for different biomes. Numbering of plant traits corresponds with the numbering of the chapters in
the handbook

Plant trait Preferred unit Range of values Recommended no. CV
of replicates Range
Minimum Preferred (%)
2 Whole-plant traits
2.1 Life history Categorical – 3 5 –
2.2 Life form Categorical – 3 5 –
2.3 Growth form Categorical – 3 5 –
2.4 Plant height m < 0.01–140 10 25 17–36
2.5 Clonality Categorical – 5 10 –
2.6 Spinescence 5 10 –
Spine length mm 0.5–300
Spine width mm 0.5–30
Spine length: leaf length Unitless 0–30
2.7 Branching architecture No. of ramifications 0 – > 100 5 10 –
per branch
2.8 Leaf to sapwood area Unitless 100–103 5 10 –
2.9 Root-mass fraction Unitless 0.15–0.40 5 10 –
(for seedlings,
down to 0.10)
2.10 Salt-tolerance traits 5 10
Selective root cation uptake Unitless – 10 –
Salt excretion and compartmentalisation Categorical – 5 –
2.11 Relative growth rate mg g1 day1 2–300 10 20
2.12 Shoot flammability Unitless 0 – ~3 5 10
2.13 Water-flux traits 10 20
Gap fraction Unitless 0–1
Stem flow % 0–50
Water retention on plant surface g m2 0–500
Leaf wettability degrees (contact angle) 0–180
Droplet retention ability degrees (slope angle) 0–90

(continued next page )

Journal compilation  CSIRO 2016

716 Australian Journal of Botany N. Pérez-Harguindeguy et al.

Appendix 1. (continued )
Plant trait Preferred unit Range of values Recommended no. CV
of replicates Range
Minimum Preferred (%)
3 Leaf traits
3.1 Specific leaf area m2 kg1 (mm2 mg1)A < 1–300 5, 5 10, 4 8–16
3.2 Area of a leaf mm2 1 – > 206 5, 5 10, 4 17–36
3.3 Leaf dry-matter content mg g1 50–700 5, 5 10, 4 4–10
3.4 Leaf thickness mm < 0.1–5B 5 10
3.5 pH of green leaves or leaf litter Unitless 3.5–6.5 5, 5 10, 4 1–6
3.6 Leaf nitrogen and phosphorus
Leaf nitrogen concentration mg g1 5–70 5, 5 10, 4 8–19
Leaf phosphorus concentration mg g1 0.2–5 5, 5 10, 4 10–28
3.7 Physical strength of leaves 5, 5 10, 4 14–29
Force to tear N mm1 0.17–40
Work to shear J m1 0.02–0.5
Force to punch N mm1 0.03–1.6
3.8 Leaf lifespan and duration
of green foliage
Leaf lifespan Month 0.5–200 5, 40 10, 160 11–39
Duration of green foliage Month 1–12 5 10 –
3.9 Vein density Mm mm2 0.5–25 5 10
3.10 Light-saturated photosynthetic rate mmol m–2 s–1 2–30 5 10
3.11 Leaf dark respiration mmol m–2 s–1 0.4–4 5 10
3.12 Photosynthetic pathway Categorical – 3 3 –
3.13 C-isotope composition ‰ –35 – –10 5 10
3.14 Electrolyte leakage % 2–100 5, 5 10, 4 9–26
3.15 Leaf water potential MPa –7!0 5, 5 5, 10 11–33
3.16 Leaf palatability % Leaf area consumed 0–100 10 20
3.17 Litter decomposabilityC % Mass loss 0–100 10 20 7–14
4 Stem traits
–3 –1 A
4.1 Stem-specific density mg mm (kg l ) 0.1–1.3 5 10 5–9
4.2 Twig dry-matter content mg g1 150–850 5 10 2–8
4.3 Bark thickness mm 0.1 – > 30 5 10
4.4 Xylem conductivity 5 10 21–63
Stem-specific xylem hydraulic kgm–1 s–1 MPa–1 1 (gymnosperms)
conductivity (KS) to 200 (tropical lianas)
Leaf-area-specific xylem hydraulic kgm–1 s–1 MPa–1 6  105
conductivity (KL) (gymnosperms)
to 1  102
(tropical lianas)
4.5 Vulnerability to embolism (y50) MPa –0.25–14 5 10 20–45
5 Below-ground traits
5.1 Specific root length m g1 3–350 5, 10 10, 10 15–24
5.2 Root-system morphology 5 10
Depth m 0.05–70
Lateral extent m 0.05–40
Density of exploration mm mm–3 104–1
5.3 Nutrient uptake strategy Categorical – 5 10 –
6 Regenerative traits
6.1 Dispersal syndrome Categorical – 3 6 –
6.2 Dispersule size and shape
Size (mass) mg (g)A 5 10
Shape Unitless 0–1 3 6 –
6.3 Dispersal potential Dispersules – 10 20 –
6.4 Seed mass mg 103–107 5 10 14–27
6.5 Seedling morphology Categorical – 3 6 –
6.6 Resprouting capacity Unitless 0–100 5 10 –
Alternative preferred units in parentheses.
Considering only photosynthetic tissue; total leaf thickness can be > 40mm in some succulent plants.
Replicate numbers correspond to the number of individual plants (replicates) from which to collect leaf litter; number of leaves in each sample will depend on
its weight and the size of the litterbag.

Australian Journal of Botany, 2013, 61, 167–234

New handbook for standardised measurement of plant
functional traits worldwide

N. Pérez-Harguindeguy A,Y, S. Díaz A, E. Garnier B, S. Lavorel C, H. Poorter D, P. Jaureguiberry A,
M. S. Bret-Harte E, W. K. CornwellF, J. M. CraineG, D. E. Gurvich A, C. Urcelay A,
E. J. VeneklaasH, P. B. ReichI, L. PoorterJ, I. J. WrightK, P. RayL, L. Enrico A, J. G. PausasM,
A. C. de VosF, N. BuchmannN, G. Funes A, F. Quétier A,C, J. G. HodgsonO, K. ThompsonP,
H. D. MorganQ, H. ter SteegeR, M. G. A. van der HeijdenS, L. SackT, B. BlonderU, P. PoschlodV,
M. V. Vaieretti A, G. Conti A, A. C. StaverW, S. AquinoX and J. H. C. CornelissenF
Instituto Multidisciplinario de Biología Vegetal (CONICET-UNC) and FCEFyN, Universidad Nacional de Córdoba,
CC 495, 5000 Córdoba, Argentina.
CNRS, Centre d’Ecologie Fonctionnelle et Evolutive (UMR 5175), 1919, Route de Mende,
34293 Montpellier Cedex 5, France.
Laboratoire d’Ecologie Alpine, UMR 5553 du CNRS, Université Joseph Fourier, BP 53, 38041 Grenoble Cedex 9,
Plant Sciences (IBG2), Forschungszentrum Jülich, D-52425 Jülich, Germany.
Institute of Arctic Biology, 311 Irving I, University of Alaska Fairbanks, Fairbanks, AK 99775-7000, USA.
Systems Ecology, Faculty of Earth and Life Sciences, Department of Ecological Science, VU University,
De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands.
Division of Biology, Kansas State University, Manhtattan, KS 66506, USA.
Faculty of Natural and Agricultural Sciences, School of Plant Biology, The University of Western Australia,
35 Stirling Highway, Crawley, WA 6009, Australia.
Department of Forest Resources, University of Minnesota, 1530 N Cleveland Avenue, St Paul, MN 55108, USA and
Hawkesbury Institute for the Environment, University of Western Sydney, Locked Bag 1797, Penrith, NSW 2751,
Centre for Ecosystems, Forest Ecology and Forest Management Group, Wageningen University, PO Box 47,
6700 AA Wageningen, The Netherlands.
Department of Biological Sciences, Macquarie University, Sydney, NSW 2109, Australia.
Department of Biological Sciences, Stanford University, Stanford, CA, USA.
Centro de Investigaciones sobre Desertificación (CIDE-CSIC), IVIA Campus, Carretera Nàquera km 4.5,
46113 Montcada, Valencia, Spain.
Institute of Agricultural Sciences, ETH Zurich, Universitätstrasse 2, LFW C56, CH-8092 Zürich, Switzerland.
Peak Science and Environment, Station House, Leadmill, Hathersage, Hope Valley S32 1BA, UK.
Department of Animal and Plant Sciences, The University of Sheffield, Sheffield S10 2TN, UK.
NSW Department of Primary Industries, Forest Resources Research Beecroft, NSW 2119, Australia.
Naturalis Biodiversity Center, Leiden, and Institute of Environmental Biology, Ecology and Biodiversity Group,
Utrecht University, Utrecht, The Netherlands.
Ecological Farming Systems, Agroscope Reckenholz Tänikon, Research Station ART, Reckenholzstrasse 191,
8046 Zurich, Switzerland and Plant-Microbe Interactions, Institute of Environmental Biology, Faculty of Science,
Utrecht University, Utrecht, The Netherlands.
Department of Ecology and Evolutionary Biology, University of California, Los Angeles, 621 Charles E.
Young Drive South, Los Angeles, CA 90095-1606, USA.
Department of Ecology and Evolutionary Biology, University of Arizona, Tucson, AZ, USA.
Institute of Botany, Faculty of Biology and Preclinical Medicine, University of Regensburg, D-93040, Regensburg,
Department of Ecology and Evolutionary Biology, Princeton University, Princeton, NJ 08544, USA.
Centro Agronómico Tropical de Investigación y Enseñanza, CATIE 7170, Cartago, Turrialba 30501, Costa Rica.
Corresponding author. Email:

Journal compilation  CSIRO 2013

.....214 3 Leaf traits ........215 3.............. bud banks and below-ground storage 3................... giving rise to a demand for standardised ways to measure ecologically meaningful plant traits..................................... Variation in plant functional traits............. past and future......3 Bark thickness (and bark quality) .....17 Litter decomposability ..171 water-use efficiency .3 Dispersal potential ...................................................................................1 Selection of species....1 Life history and maximum plant lifespan ......................211 2.............................14 Electrolyte leakage as an indicator of frost 2 Whole-plant traits.....216 3...................2 Root-system morphology.... Additional keywords: biodiversity.........11 Leaf dark respiration ...2 Life form .. physiological..........217 3............186 6 Regenerative traits ...................................................202 2............ affect other trophic levels and influence ecosystem properties................170 3.....................................6 Resprouting capacity after major disturbance... Abstract......................... step-by-step recipes...............182 5..........189 6................................................................... and for traits that can be measured relatively easily.................. the impacts of species invasions.....178 4.................2 Twig dry-matter content and twig drying time.......................................207 2.......192 6..201 2........168 Australian Journal of Botany N........................ and puts particular emphasis on traits important for predicting species’ effects on key ecosystem properties.........184 5.........6 Leaf nitrogen (N) concentration and leaf phosphorous 6..16 Leaf palatability as indicated by preference by 2....170 3..199 1..............13 C-isotope composition as a measure of intrinsic 1.178 4...and ecosystem- level processes.................. published online 26 April 2013 Contents 3.......................212 2................ with a minimum of text on theory.....................212 2..........7 Branching architecture ......3 Replicate measurements.......... leaf traits....................3 Nutrient-uptake strategy...........5 Vulnerability to embolism ....................1 Specific root length ............... and trait syndromes.. Plant functional traits are the features (morphological.....13 Water-flux traits ....173 3....................216 3.. this handbook retains the focus on clearly presented................... ecophysiology.................... ecosystem dynamics............................... has proven useful for tackling many important ecological questions at a range of scales.. plant morphology................15 Leaf water potential as a measure of water 2......4 Seed mass................................................................................181 5 Below-ground traits .......179 4................................200 1......................................... This line of research has been among the most fruitful avenues for understanding ecological and evolutionary patterns and processes........179 4....................... We hope this new handbook becomes a standard companion in local and global efforts to learn about the responses and impacts of different plant species with respect to environmental changes in the present.....218 (P) concentration............................. Received 23 November 2011.....220 ......................4 Leaf thickness ..................................195 References.177 4..172 3..................12 Plant flammability....................................6 Spinescence.............205 organs....220 3................8 Leaf area : sapwood area ratio ........ Updated and expanded from the widely used previous version.....................11 Relative growth rate and its components....... in particular for traits with power to predict plant.176 4 Stem traits ...........203 2.........4 Plant height ....................................190 6.. It also has the potential both to build a predictive set of local.....................................5 Seedling functional morphology.................. phenological) that represent ecological strategies and determine how plants respond to environmental factors............. and increases the value of standardised protocols for quantifying trait variation of different species..........................................209 2..............1 Specific leaf area .....207 2...............................12 Photosynthetic pathway ............198 1 Selection of species and individuals ..5 Clonality.............. environmental change......................9 Vein density ....................193 Acknowledgements.............214 3......4 Xylem conductivity.........173 status ..........191 6..... The importance of these topics dictates the urgent need for more and better data....................................190 6............................. This new handbook has a better balance between whole-plant traits......... widely applicable.......................172 3.........175 model herbivores.....................5 pH of green leaves or leaf litter .......172 sensitivity ......3 Growth form ............ alterations in biogeochemical processes and vegetation–atmosphere interactions.................... but also introduces many new protocols for further traits....8 Leaf lifespan and duration of green foliage ................10 Light-saturated photosynthetic rate ..............197 3.. including changes in biodiversity............... accepted 29 January 2013.................................3 Leaf dry-matter content..........................................................................................................................7 Physical strength of leaves.......................210 2.........198 Introduction and discussion ...... regional and global relationships between plants and environment and to quantify a wide range of natural and human-driven processes.........................2 Dispersule size and shape ...........1 Dispersal syndrome....................... root and stem traits and regenerative traits...............169 3..............2 Area of a leaf ........................ and not only includes updated methods for the traits previously covered...........1 Stem-specific density ...............215 3............................... Pérez-Harguindeguy et al.......................9 Root-mass fraction ................. ecosystem functions...........10 Salt resistance......2 Selection of individuals within a species....186 5.......218 3........209 2.

Cornwell et al. there has been a rapid expansion of large regional and factors. associations of plant traits). easily implemented trait-measurement recipes for more important. Paula that variation in plant traits.try-db. properties (Kattge et al. environment (Lavorel and Garnier 2002). 2004. Díaz et al. Aerts and Chapin 1999. Violle et al. inexpensive and standardised modifications in climate at the global scale has been measurement in many biomes and regions. and yet informative enough to composition. 2003) was written to address that need. we consider plant functional traits to be that global collective initiative. 2009. associated with many important ecological processes at a see Box 1) is compiling a communal worldwide database of plant range of scales. (2007). Ample evidence concentrations as an indicator of both potential rates of indicates that plant traits and trait syndromes significantly metabolism and of quality as food for herbivores) to plant affect ecosystem processes and services (for overviews. Cavender-Bares practical ones related to nature conservation and land et al. 2010. Lavorel and Garnier 2007. nucleodiversus. 1993. 2010). 2004. Reich et al.g. Various complementary protocols for specific plant (eco-)physiological as well as environmental measurements not covered in this handbook can be accessed through the fellow project: Prometheus Wiki (Sack et al. 2007. 2010. an unprecedented step in improving the capacity of the standardised ways to measure ecologically meaningful plant scientific community to access and utilise plant-trait information. 1999. As proposed by Chapin et al. 2010. 2008. at the cell to the whole- with strategies and trait syndromes across floras. 2012). within and among species. see functions themselves (e. unambiguous use in any yet incorporate continuous variation in plant traits among plant terrestrial biome. decomposability.e. et al. However. Srivastava et al. Stich et al. Faith et al. step-by-step et al.g. Harrison et al. leaf nutrient Ordoñez et al.g. The traits contained in the handbook represent a consequence.csiro. study of the plant features (traits) that reflect species ecological As a consequence of this surge of theoretical and practical strategies and determine how plants respond to environmental interest. current-generation DGVMs do not recipes for direct and. Doherty and predict them has given new stimulus to a long tradition of et al. www. Lavorel et al. 2006. atmospheric at the regional and global scales. Díaz et al. Zanne et This has resulted in strong demand for traits. 2009. Edwards et al. researchers worldwide. 2004. 2008. always measured at the 2008. 2010). recurrent et al. 2008. physiological or phenological feature. Jones et al. dispersal and large-scale unprecedented ecosystem changes. standardisation of protocols applicable under a et al. Baraloto et al. 1997. 2008. and at the from local plots to landscapes to biomes. 2010). species or types (Cornwell et al. Baraloto et al.publish. 1997. land use and biotic exchanges are triggering relate to biogeochemical dynamics. To submit corrections. by providing wide range of situations and geographical contexts becomes even standardised. Cornwell Functional traits addressed in the present handbook range et al.php). 2007) or its and has attracted increasing interest in recent decades (e. et al. conservation (e. 2004. Kattge et al. To calculate functional diversity metrics and indices with your trait data: FDiversity Free Software Package (Casanoves et al. affect other trophic levels and influence ecosystem global trait databases (e. 2010a. Díaz et al. which potentially affects its fitness (cf. McGill ecosystems has been a long-standing focus in plant significantly improved with the use of dynamic global This is a recipe book to be used in the field and in the vegetation models (DGVMs) (Cramer et al. we have had to make hard choices.g. Mace plant effects on ecosystem processes and services at various scales et al. Useful links for plant functional-trait workers To find on-line protocols and updates related to this handbook: Nucleo Diversus/Tools (http://www. The need to understand disturbance (Ollinger et al. http://prometheuswiki. 2007. The identification of general plant trait trade-offs associated measurable for individual plants.g. Arneth laboratory. Ma et al. as far as possible. De Bello et al. Westoby modality taken by the trait at any place and time an ‘attribute’. (2) can help answer interface between the evolution and ecology in communities questions of ecological and evolutionary theory as well as and ecosystems (e. Chave et al. we will call the particular value or Cornelissen et al. and contains comprehensive.fdiversity. from simple indicators of plant function (e. capacity to resprout after a fire). trait-based approaches are currently also gaining set of functional traits of vascular plants that (1) can together momentum in the fields of agronomy and forestry (e. 2011. 2001. 2009). 2011. The TRY Initiative (Kattge et al.g. 2010. Brussaard represent key plant responses to the environment as well as key et al. Wright et al. There is mounting evidence Kleyer et al. detailed. Freschet et al.New handbook for measurement of plant traits Australian Journal of Botany 169 Introduction and discussion syndromes that are simple and general enough to be assessed Environmental changes such as those on climate. management (see Box 2 for a Discussion) and (3) are in most The quantification of vegetation changes in the face of cases candidates for relatively easy. 2010. with an even broader scope. traits. 2010. additions and comments to improve this handbook: traitshandbook@gmail. 2011). To that end.nucleodiversus. Wright et al. taxa and organism level. Garnier and Navas 2012). Patiño et al. Lavorel et al. 2012). Chapin et al. 2010. Fortunel et al. To share plant functional-trait data with other researchers (both as a provider and as a recipient): TRY Worldwide Initiative and Database (Kattge et al. is 2012. Next-generation models We did not intend to provide a comprehensive list of all traits that could benefit from the incorporation of functional traits and could potentially be measured nor a thorough description of the Box 1. Grime et al. 2011). 2009.g. The predecessor of the present handbook (Cornelissen In this context.g. 2010. 2012). . Cardinale et al. and trait syndromes (i. This updated version is an extension of In this manual. 2008. 2010a. As a species level. archaeobotany (e. 2002. 2011). 2011. any morphological. www.

regional or global scale (e. should help identify the most appropriate traits to measure. 1997. We hope that the focus on practical techniques comparative efforts. rather than setting limits to researchers’ curiosity. so that the next edition will be an even better bed-side for each trait. dispersers or herbivores? Does the target process occur above or below ground? Is the focus on coarse differences across or among regions or continents or on subtle differences among individuals of two slightly different local populations? Are specific ecosystem services to people assessed or predicted? All these and further types of questions will have a direct impact on the selection of traits. Reich and provides protocols for several additional plant functional et al. A discussion of these is beyond the scope of the present manual. 1998. Some examples of additional interesting traits not covered here are in the introductory text of Cornelissen et al. 2007). methodology and existing large datasets. Beyond this. on significance. the persistence of adults? Does it involve pollinators. useful reference in laboratories and in the field for studies around Particular emphasis is given to recipes appropriate for areas with the world. Which traits to measure to answer which questions? No methods handbook can answer the question of what are the best traits to measure. there is not much point in comparing multiple species for succulence within wet environments or for flammability within areas that burn only very rarely. and especially the list of references provided for each trait. and offer the possibility of comparing distant ecosystems with very little taxonomic overlap (Reich et al. relatively novel methods are described. or if the project involves an intensive one-off measurement campaign carried out by highly trained specialists or recurrent measurements by third parties. 2004. especially for organs other than the leaf.1 Selection of species and many additional studies have made to the theory and Study objectives will always determine which species are selected measurement procedure for each trait. a small number of traits have been considered relevant almost universally.g. Rather. For a particular question. seed size (usually expressed as seed mass) and the structure of leaf tissue (often expressed as specific leaf area or leaf dry-matter content). Díaz et al. 2000. It has better 2004. and under the heading Special cases or extras. We hope that the authors of relevant in Appendix 1. including ‘new’ traits. Weiher et al. Lavorel and Garnier 2002. Díaz et al. the brief ecological background. We give only brief ecological background (see Box 1). 2004. because they are at the core of the plant life cycle (Grime et al. Westoby et al. and on practical circumstances. They also provide understanding of how functional diversity in the broad sense underpins ecosystem processes and the benefits that people derive from them (Chapin et al. herbivory. Knevel et al. the specific leaf areas and stem-specific densities of these species might serve as less precise but still useful proxies for broad patterns of variation in growth and vegetation productivity. made for clarity and brevity. suggested numbers of replicates for all traits are given recipe descriptions. The recipes provided here. 1997. This new handbook both updates theory. This section presents guidelines for selecting species and Readers can find complementary methods and additional individuals within species for trait measurement. Wright et al. Pérez-Harguindeguy et al. In addition. Hodgson et al. methods and and for identifying general strategies or syndromes of resource databases covered by its predecessor (Cornelissen et al. regeneration. Gubsch et al. 2008). These are plant size (usually expressed as height). to help answer exciting novel questions. for trait measurement. (see Box 1). Specific citations have not been included in the In addition. species or populations from a broad coverage of (1) measurements important in less studied biomes range of environments and phylogenetic groups should be . For species-level analyses of trait variation. 2003). 2011). whereas for others. 1999. 2003). Why measure plant traits and which traits to measure? Plant functional traits give better insight into the constraints and opportunities faced by plants in different habitats than does taxonomic identity alone (Southwood 1977. (2) floras with special adaptations and (3) plant consensus traits and methods that researchers have identified as functions related to carbon and nutrient cycling. Logistic and financial considerations are equally relevant. theory behind each trait. as well as discussions and comments in specific associated web pages general considerations of the necessary number of replicates. McIntyre et al. The first and foremost criterion in deciding what traits to aim for is the process of interest. Westoby 1998). and in full recognition of the important contribution that each of them 1. publications (most of them cited at the end of each recipe) will understand this choice. Some of them are well known and widely and streamlined trait recipes will help this handbook become a used. use. traits. if resources are limited for measuring relative growth rate on hundreds of species representing a large gradient of productivity. because this strongly depends on the questions at hand. we 1 Selection of species and individuals give pointers to interesting additional methods and parameters.g. or trade-offs at the local. there are some ‘core lists’ of plant traits that are considered important for plant resource use. with a short list of references with further details table companion. the ecological characteristics and scale of the study area. Grime 1979). (2003). should assist in making those decisions. For instance. incompletely known floras and modest us about both general issues and specific details of these protocols research budgets. Although there is no limit to the number of relevant traits in different research contexts. dispersal and response to widespread disturbances (e. The plant-trait approach often provides unique mechanistic insights into several theoretical and practical questions. 1999. 1997. soil nutrient or fire regimes? Is it about vegetation feedbacks to atmosphere and climate? Does it involve the juvenile stage. and readers are referred to these papers for a first introduction. reproduction or dispersal over the landscape? Does it involve plant survival in response to resources or disturbance? Is the main question about response to or effects on water.170 Australian Journal of Botany N. Díaz et al. this trait handbook aims at inspiring others to come up with and measure traits not covered here. the choice of traits would be slightly different if the limiting factor is labour force or access to sophisticated analytical laboratories. For example. We strongly invite users to share their experiences with high species richness. reliable and feasible to be applied in large-scale dynamics and fire. The main section of each recipe contains a brief. 1999. water being useful. although it is not necessarily less laborious or less expensive than other methods. whereas such data might be useful as a reference in larger- scale studies. including the sections on Special cases or extras. Cornwell et al. Similarly. Is the intended study about fundamental plant or organ design in response to environmental variation in the present or about the evolution that gave rise to today’s spectrum of designs? Is it about plant growth. the present handbook contains and ecosystems. Box 2. standardised protocol.

standard deviation cycling). Gaucherand and Lavorel 2007). should be selected. in the understorey of closed forests. Exceptions may be made if this criterion would imply from different environments. whether ecosystem level).g. e. following Garnier et al. species should be selected quantitative traits described in the present handbook. gradients. Taxon-free have to be measured when they are present and fully developed. or how vegetation characteristics Swenson 2010. ‘healthy-looking’ plants for trait measurement (see show some variation within species along environmental Section 1. Although most traits robust. with may therefore vary in which quantitative traits are stable across little consideration about their abundance in situ.g. 2009). and their importance locally Lavorel et al. a light gradient (cf. Moreira et al. Species criteria (such as being members of particular clades). Traits known to often be neighbouring vegetation. pollution. it may be useful to distinguish ‘variable’ traits from from those obtained using the standard approach of selecting more ‘stable’ traits (Garnier et al. especially combined with a very high evenness. The reverse is the 1. (‘trait-transect’ method. communities with unusually high species richness per unit bears an uncertainty (and CV will likely increase as scale area. e. Hulshof and vegetation characteristics. Messier et al. of nutrients or disturbance. This does questions and associated traits (Lavorel et al. By unless specific goals suggest otherwise. found. it should be that the relative abundance and the traits should be measured at noted that different methods are relevant to different ecological the time of peak standing biomass of the community. condition across the habitat range (Garnier et al. ‘stable traits’ can be measured for any representative with the light environment. dispersal and pollination modes. In predominantly herbaceous systematically. we suggest ecology and vegetation-science textbooks.g. mineral nutrient concentration in leaves. Examples increases). however. 2008). see also not always apply to reproductive structures. Some quantitative traits such as be collected from the least shady places in which they still look . Leaves of these species could photosynthetic type (C3 or C4). along a transect level properties across environmental conditions (e. as well as for the ease with which they can be been applied to tropical forests. 2010.e. hereafter CV) for some of the local abundance.New handbook for measurement of plant traits Australian Journal of Botany 171 selected.g. by looking at the range of CVs calculated are tropical rainforests and fynbos vegetation. Methods of taxon-free sampling have also and regionally. which implies that they For robust comparisons across species. onset of flowering. indicator species individuals rooted nearest to random sampling points. each of the individual estimates e. capture the contribution of more abundant species. branching architecture and spinescence. measuring traits regardless of species identity. primary and Appendix 1 gives a rough indication of the within-species secondary production. We. approaches that do not require species identification offer an which sometimes does not coincide with the time of maximum alternative to estimates of relative abundance. (2004) and Pakeman and that can be expected. select points or quadrats at random or of the overstorey woody species. Trait values obtained through these methods can differ sense. 2011). often plants located in well lit ‘variable’ include vegetative and reproductive plant height. Because of the low number of measurements for an impracticably large numbers of species. For questions about evolution. or to follow a different method) is beyond the communities.g. 2010b. or very close to the clonality.g. replicates generally used. 2012.2 Selection of individuals within a species case for so-called ‘variable traits’. affect local flows of matter and energy (e. 2012). or on other phylogenetically relevant along others. This is particularly important for some leaf traits (see Section 3. or leaf toughness can be ‘stable’ be based on the inclusion of representatives of different enough along certain gradients. e. which may strongly depend on population from the entire gradient. a reasonable estimate of the 100 species per plot may be needed to reach the 80% biomass typical within-species-at-a-site variability can be obtained. abundant species of the understorey may also be included (e. the most the CV distribution. De Bello et al. preferably totally unshaded. in this case. therefore.1). 2009. given gradients. or in response to specific environmental changes. Poorter et al. threshold. or for or different regional climate or fertility levels). as long as can be selected on the basis of the sensitivity of their trait values to the canopy structure is quite simple (‘trait-random’ method – the environmental factor of interest. However. which obviously Baraloto et al. 2010. environments. present in Appendix 1 the 20th and 80th percentiles of In forests and other predominantly woody vegetation. strongly based found and identified in the field (independent of their relative on the frequency or basal area of individual trees (Baraloto et al. along that collectively make up for ~80% of cumulative relative with the more frequently used units and the range of values abundance. How species abundance should be measured to determine the when the research question relates to the whole-community or species making up 80% of cumulative abundance (e. and to a lesser degree ground in multilayered grasslands. so tests should be made before a trait is taken to be when trying to understand how environmental variables shape stable for a given species (Albert et al. the main criterion for species selection should be divided by the mean.g. Traits that are This criterion creates sampling problems for true shade species relatively ‘stable’ include categorical traits. carbon.2). and effectively vegetative growth. species contribution to a particular community may scope of the present handbook and is extensively covered in plant- vary with time during a growing season. traits should be generally should preferably be measured in more than one site or measured on reproductively mature. abundance) (Ansquer et al. water and mineral nutrient variability (coefficients of variation. healthy-looking individuals. even if their biomass is much lower than that to lay out transects. the intraspecific variation of so-called ‘stable traits’ is low compared with their interspecific variation. the choice of species may leaf and stem dry matter content. 2007). 2007). In this 2010b). i. being. As a first step. 2008. In contrast. These include For comparing sites or for monitoring trends in ecosystem. In those cases. such as life form. in which well over across a wide range of species. specifically for tropical forest).g. Appendix 1 summarises field data Quested (2007) (see specifics for abundance measurements collected in several studies for a wide range of species coming below). To avoid interaction contrast. but not phylogenetic groups.

low temperatures and feasible. Qualitative distinction between life-history classes species v. followed by number of species covered. the crown will 2. e. there should be individuals with a storage root but not an inflorescence. e. compared with that among species means. rhizome each species. as well as for variance component analyses (used to partition variance among different levels. A formal analysis hemicryptophytes) near or below ground surface. except that perennial wild plants plants but may be apparently nearly unlimited in clonal plants. Pérez-Harguindeguy et al. plants in the absence of disturbances or catastrophes. If biennial. the study concerns just a small number of species or a modest in the next season. This and produce seeds or fruits with low dispersal potential. mainly based on common practice. Other more powerful techniques can also A plant with any perennating organ other than the seed is either be used. within or between ecosystems or geographical regions. Plant grows vegetatively the first season. 2010. such as mixed models (Albert et al. Appendix 1 shows senesces and dies. from one growing season among individuals. because to short-lived species. It is highly recommended to quantify the relative or ‘root crown’ (bud-bearing stem base or contributions of intra. stem has lateral Ideally.1 for a (a) Monocarpic perennials. for consistency among sets of measurements. so the fundamental unit on which a trade-off between maximum lifespan and dispersal in time and measurements are taken should be the ramet. After several to many seasons discussion on within-species variability). researchers should check within-species CV at their site thickening over the years. its thickness extending continuously of population persistence and is therefore strongly related to into the stem. the fewer replicate measurements can senescence and death of the shoot and root system. Plant senesces and dies at the end of its first 1. and so has two seasons.v. some living.172 Australian Journal of Botany N. Messier et al. be aware that How to assess? sampling of neighbouring ramets may not provide biologically independent replicates for species-level statistics). and others with both. Long-lived species often exhibit a short-lived seed bank a recognisably separately rooted. usually do not flower in their first year. above-ground shoot. one may need to sample more heavily within perennating organ such as a bulb. In broad-scale interspecific studies. the greater the flowers in the second to produce seed. after producing seed. Aerial shoots (and sample relatively few plants of any given species. A plant that 2 Whole-plant traits lacks specialised perennating organs may still be perennial. then precision.1).
 of statistical power based on an assumed or known variance (ii) Woody perennial retains. Maximum plant lifespan is an indicator relatively soft and smooth. individuals). frequency is mostly negative. The relationship with disturbance same individual to measure as many different traits as possible. in contrast choice is both pragmatic and ecologically sound.5). the root of an annual is usually respective individual. be made for each species. Moreira et al. species variability in the trait of interest (see Section 1. 2012).g. which Trait values are often used comparatively. If environmental stress regimes. by resprouting from its root-crown. This type although the first may be very short). Plants severely Maximum lifespan is strongly positively associated with affected by herbivores or pathogens should be excluded. whereas an annual always . the ramet is likely to be the unit of most interest for most functional. corm. Individuals for (A) Life history measurement should be selected at random from the population This simple classification distinguishes among the common of appropriate plants. then number or range of species to be sampled. There may be species (see Section 2. The individual survives for at least three selected for measurement should depend on the natural within. whereas when sometimes roots) die off as growing season ends. which often have a very long-lived seed genets are often difficult to identify in the field and. defined here as space. the plant produces seeds. in contrast. of research almost inevitably implies a conflict between scale and (2) Biennial. one may 
 (i) Herbaceous perennial. trait-related questions (however. given constraints of time and labour. interspecific variation. Commonly used statistical packages generally include which die by the end of their third season or later. leaf-bearing shoots. be used. as well as on the of vegetative growth. case. and can eventually become quite thick and period from establishment until no live part remains of the even woody (a caudex).1 Life history and maximum plant lifespan normally carry wrinkles or scars from bud outgrowth in Plant lifespan (usually measured in years) is defined as the time previous seasons. All or much of the stem and traits. A perennial in its first year of growth may resemble land use and climate change. (1) Annual. or by using a systematic transect or quadrat types of timing and duration of survival behaviour of individual method. can into the next. although long-lived (resprouting) Defining ‘individuals’ reliably may be difficult for clonal clonal plants may also tolerate frequent disturbance. before deciding this. growing seasons. Lifespan is limited in non-clonal an annual in these respects. routines for power analysis. in any bank and/or high dispersal potential.3 Replicate measurements growing season (from seed).g. the minimum and preferred number of replicates for different (b) Polycarpic perennials. The most appropriate root system normally survives the harsh or dormant sample size depends on the purpose and scope of the study. The number of individuals (replicates) (3) Perennial. to classify species into may propagate a new plant in the future (a winter annual different functional groups or to analyse variation across species germinates in late summer or autumn. If so. use the low nutrient availability. new shoots grow from a local gradient. 2010. a perennial or a biennial (the latter only by a storage taproot). healthy and not discoloured (see Section 3. period between growing seasons.

(2003). If a ramet never becomes independent intermediate forms. Maximum lifespan within a population is studied Schweingruber (1996). whereas annual shoot-length increments ones. microscopic cross-sections are affect canopy structure. Fischer and Stöcklin (1997). e. with stem and root tissues that are decomposed. the heather Cassiope tetragona. boreal or austral. chryosidine. Category C under Section 2.2 Life form clear observation. for most shrubs where single shoots have a limited age. (B) Maximum plant lifespan quantitative assessment (3) Cold-climate dwarf shrubs. annual rings in the tap root (e. information is given in Material S1. at or very close to the soil surface.5 for a definition of (2) Geophyte species. with rosettes and prostrate growth forms being counted among all samples (although the mean lifespan of all associated with high grazing pressure by mammals. Maximum lifespan of a species or positioning of the foliage to avoid or resist grazing by particular population is defined as the largest number of annual rings herbivores. lateral annual rings are often In gymnosperms and angiosperms. a study on 900 temperate herbaceous species revealed the annual sequence of distances between leaf scars. Leaves concentrated on a short. However. sometimes be absent. astrablue associated with ecophysiological adaptation in many ways. The annual rings can usually be counted under a Plant life-form classification sensu Raunkiaer (1934) is a simple dissection microscope. De Witte and minimum of 10. dwarf shrubs). any of these categories.New handbook for measurement of plant traits Australian Journal of Botany 173 does (many horticultural perennials. species maximum lifespan can be estimated by counting the of woody stems can be distinguished under a microscope number of annual rings representing annual tissue increments. Because we are classifying types along a continuum. However. and both the vertical essential and have to be treated first by ‘eau de javelle’ to remove and horizontal distribution of leaves. more complicated. Data are collected from a (2001). may rhizome). In herbaceous species. We. In some cases. Often a cross-section of a shoot does not but still a useful way of functionally classifying plants. shrubs. It is important to obtain a rather smooth surface for 2. in clonal plants where the genet rather soft compared with typical wood. Life history varies with location species. These collar. recommend digging out woody plants a bit and taking 2. In the latter case. and wet season. In the two latter climate zones. and any branching of the main-shoot axis or axes. In woody Stöcklin (2010). Growth form may be the cytoplasm and then stained (fuchsin. only permanent-plot research to do so). Schweingruber and Poschlod (2005). (1980). annual rings may the same region. (FCA). faster- with clear seasonality (cold (winter) or drought seasons) such as growing species fall into different life-history categories in the polar.g.3 Growth form (additional) samples from the root collar. with an disappear above ground for up to several years before inflorescence (or single-flower peduncle) bearing . mechanically and nutritionally self-supporting albus) are also a suitable tool to identify maximum lifespan plants of a genet. especially monocotyledons. through the winter-mark septa separating them and through Recently. available as Supplementary stem transition zone of primary roots). In particular. including its height. however. species (trees. the formation of annual rings can depend on and should preferably be assessed in the field rather than by habitat conditions. polarised light has proven to be useful from severe climatic conditions. annual rings may even be found in tropical species. Dictamnus (A) Terrestrial. either by cutting out a whole cross-section or a ‘pie slice’ of the Cherubini et al. many short-lived. temperate and even in Mediterranean. and also in rhizomes. have been selected reappearing. with individually marked individuals will give an idea about the maximum lifespan of those species. alternatively. (2009).g. Here. Gatsuk et al. even within type zones. maximum lifespan can be (1) Herbaceous plants have either no or at most modest higher if part of the rhizome or stolon is already secondary growth. as well as occasional unique forms lying outside plant. consists of more or less independent ramets. the identification of (maximum) lifespan is literature. may from the genet and will never be released from the mother be encountered. be estimated only indirectly by means of size or diameter of condensed section of stem or rhizome (see a genet in relation to mean annual size increment. annual rings are determined More on methods: Tamm (1972).g. sheltering rings visible. individuals may be informative too). Armeria maritima. which is especially true Material for this paper. even in some non-woody very hard to discern. or by taking a core with a pole-testing drill (tree corer). Growth form is mainly determined by the direction and extent of annual rings are mostly found at the shoot base or in the root growth. different regions and a few differ among habitats. How to record? Special cases or extras Growth form is a hierarchical trait assessed through field observation or descriptions or figures or photographs in the (1) In clonal plants. Larson in the largest and/or thickest individuals. or optimising the height and to identify the annual rings. annual rings in perennating structures in more than 80% of the (4) Life history and location. Rozema et al. Silene acaulis) or annual morphological markers along the rhizome or stolon (e. In some of these species. astrablue and safranin) to make the annual including maximising photosynthetic production. preferably 20 individuals (replicates). More represent the maximum age as precisely as the root collar (root. between the categories recognised here. therefore. In some cases. genet age can (a) Rosette plant. especially in regions with a distinct dry References on theory and significance: Rabotnov (1950). In such cases. main stem (trunk). Lycopodium annotinum. Annual rings will be found in vegetation zones reference to floras.

above ground level. An excurrently branched (cf. by sclerification to support a vertically extensive.g. Many individual shoots of a dense upward to produce a wider. soft..3.g. the top. (b) Woody vine. Many forest cotton grass. (a) Palmoid. it later becomes . Morphology as in one of Types (i) living shoots with active leaves (e. Point leaf positioning. palms (Pandanus). most leaf succulents fall However.g. Pteridium aquilinum (bracken fern). Cymbalaria muralis). somewhere developed inflorescences. Eriophorum vaginatum). soft or ‘anomalous’ secondary vegetation. normally without contact with the Espeletia spp. Plant that grows in or on rocks (e. with main foliage held close to soil surface. or (epiphytes) on an above-ground support ascending. unshaded habitats. Long-lived woody stem individually up into the light. Stem without secondary growth such as ‘pine barrens’. (c) Cushion plant (pulvinate form). without any means of (c) Stem succulent. either no or reduced leaves (bracts) produced from instead into one of the subclasses of Points A. Aerial inflorescences (or (c) Shrub. Utricularia their stems undergo more extensive secondary forestii. arborescent ground (e. Pérez-Harguindeguy et al. (2) Semi-woody plants. species of Nepenthes. Iris (b) Dwarf shrub. but also in various climatically or nutritionally challenging. Usually attaches to its support having only weak secondary growth is stiffened either by twining or by means of tendrils. Woody plant between 0. and some Bombacaceae can be (2) Lithophyte.
 have this form). directly bears photosynthetic leaves that extend (a) Prostrate subshrub. cycads. include many Arctic willows and ericoids). semi-deserts. stem(s) whose nodes bear photosynthetic leaves ascending or horizontal branches giving a that are distributed nearly throughout the canopy of conical or (in mature trees) columnar form the plant. stem with extensive. Graminoids (rhizomatous above its base. at least references below). many tropical orchids and Bromeliaceae).g. Viola spp.g.i in the present Section) trunk lacking or (a) Herbaceous vine. the Arctic or (ii) but substantially <5 m tall. Plant that grows attached to the trunk or branched stem (e. or none. growth (see also ‘Corner model’ within the (3) Climber. understorey trees.d. less equal branches that continue branching (e) Tussock. (i) Excurrent. The and corky outer bark from cork cambium (woody vines permanent axis is an elongated rhizome that are covered in Point B. unbranched or little. trunk. mostly dead supporting column topped by (e) Dwarf tree. usually tough. various support by means of aerial roots. and lacking in distally (ii) Deliquescent. with usually multiple.). tall. except when shed from its more basal to the crown. grasses’) fall in this category. spp. many regarded as having this growth form. Dracaena. Bears a rosette-like canopy of typically (B) Plants structurally or nutritionally supported by other plants large.1 or the rosette axis. on external support for its upward growth and (b) Bambusoid. Woody plant usually >5 m tall. certain but often toughened by sclerification (or. May start epiphytically (but become only limited secondary growth (cacti.). woody stems less than 0. leaving behind a crown.3 in the present Section). such as a tree branch. whose principal photosynthetic leaves are attached to the base of their aerial stems (e.g. water-storage tissue and (d) Strangler. bogs and near-timberline with relatively feeble. secondary xylem and phloem from vascular cambium. more flat-topped colony or clone grow upward. by secondary growth. including liana. although species of ferns. Trunk divides. Galium spp.g. single-flower peduncles) with either reduced with canopy typically carried by several trunks that leaves (bracts). Yucca spp. (c) Scrambler. tropical cloud forests.174 Australian Journal of Botany N. tree trunks. tree ferns). substantial. tough. (b) Elongated. (1) Epiphyte. Single main axis (trunk) extends (d) Extensive-stemmed herb develops elongated aerial up to. A.5 m tall.5 m and ~5 m tall. Plant that roots in the soil but relies. ‘bunch (3) Woody plants develop extensive. leaf-bearing rhizomatous. may grow out from the are usually thinner and younger than typical mature rhizome. herbaceous dicots such as Chenopodium. branch of a shrub or tree (or to anthropogenic supports) Certain tropical or alpine Asteraceae such as by aerial roots. or almost to. The rhizome can growing horizontally at ground level (examples be located either at or below ground level (e. and cactoid soil-rooted) or by climbing from ground level. canopy of other plants. or subshrub. with shorter. more or or not) with leafy aerial stems fall here. A usually leafless photosynthetic attachment (e. columnar. often compound leaves atop a usually thick or by special physical features (‘pachycaulous’).. alternatively. even and rounded canopy form (many alpine plants usually single (but upwardly branching). plants of other families. Tightly packed (d) Tree.
 parts during later growth. Graminoids A. with relatively canopy elevated on a long-lived.3 of the present Section). into two to several. Often attaches to a sometimes tree-sized canopy (bamboos. Grows up through a sufficiently dense Amaranthus and Helianthus). growth). initially.

significance and large datasets: the stem should preferably equal 1–1. with growth form. or the projection of an inflorescence above the vegetative part maximum height (Hmax). plant height is based on the rosette leaves. Additionally. The height recorded should correspond (1996).1 in the present Section.4 Plant height height may be somewhat tricky to measure (if the plant bends. McIntyre et al. leaves or photosynthetic portions of the initially supporting stem (e. depending on species size and the number from eye to a location at the crown margin that is level with of plants and time available. vegetative plant 2. Cain (1950). this is preferably carried out towards the end of More on methods: Korning and Thomsen (1994). position of the species in the vertical light while recording maximum canopy height. Medina (1999). Hirose and Werger (1995). arbitrarily use a gradient of the vegetation. For herbaceous et al. is the maximum stature a typical mature of the plant may be a useful trait in responses to disturbance.New handbook for measurement of plant traits Australian Journal of Botany 175 self-supporting. Ellenberg and Müller-Dombois (1967). to the top of the general canopy of the plant. competitive vigour. Because plant height is quite stem. Whittaker (1975). individual of a species attains in a given habitat. Box (1996). (2) for tall tree species. storm. the formula to calculate the height of the thickest individual in the stand. (2) Herbaceous. (1999). the angle between the of Point A. King the main stem if this projects above the foliage. and references therein). Special cases or extras Richter (1992).5 times the tree height. or use must be placed vertically at the base of the tree. For herbaceous species. Ewel and Bigelow (1996). not the height of the inflorescence (or seeds. proportionally very little photosynthetic area higher up. Lüttge (1997). Others. it is easier to measure (with an variable both within and across species. (5) Dense undergrowth. it is difficult to estimate the height above the tree light exposure for that species). For such individuals. vertical height from eye level down to tree base (a subtraction height measurements are time-consuming. significance and large datasets: Gaudet The height to be measured is the height of the foliage of the and Keddy (1988). and aquatic plant that relies on surrounding water for (2) trigonometric methods such as the measurement of the physical support.) horizontal plane and the tree top (a) and between the (5) Parasite or saprophyte obtains important nutritional horizontal plane and the tree base (b). disturbance events (such as e. discounting any (1999). Poorter et al. whereas reproductive plant height can be ‘safer’ of the main photosynthetic tissues (excluding inflorescences) on a in this sense. fruits). expressed in metres. certain tropical Ficus inflorescence. when more time is available. reproductive size. For epiphytes or certain hemi-parasites (which penetrate tree or shrub branches with their haustoria). or if inflorescence has significant photosynthetic Plant height is the shortest distance between the upper boundary portions). and whether a species is to estimate the position of a transition between vegetative and able to establish and attain reproductive size between two reproductive growth. McIntyre and Lavorel (1) Rosettes. including the following: (1) for the tree top. Westoby (1998). Hmax is associated so both of these heights should be useful to measure.g. Moles et al. 45 degrees (a smaller height error caused by inaccuracy in the readings). spp. 2008). (2006). they involve the use of a pole of known height that derive the asymptote from the regression coefficients. For trees with large spreading exposed to full sunlight (or otherwise plants with the strongest crowns. potential lifespan. (2003). For estimating the height of tall trees. grazing). tree height (H) is needs directly or indirectly from other vascular plants then calculated as H = d  [tan(a) + tan(b)].g. More on methods: Barkman (1988. some options are (4) Submersed or floating hydrophyte. height What and how to measure? is defined as the shortest distance between the upper foliage boundary and the centre of their basal point of attachment. Rundel (1991). ploughing. . multiply this by the sine of the sighting angle to short species. Kohyama et al. Weiher et al. Barkman (1988. or Thomas (1996). height of the five tallest mature individuals can be measured. Thomas the growing season. (2006. and references therein). species. and above. Cramer (1997). Healthy plants should be sampled that have their foliage (4) Large spreading crowns. height estimates (parasite) or from dead organic matter in the soil are most accurate if the measurement angle is between 30 (saprophyte) (see Nutrient uptake in Material S2 where degrees (easier to define the highest point in the crown) and other more specific forms of parasitism are covered). there are three ways optical rangefinder or laser) the vertical height as the distance to estimate Hmax. species. and for these. For plants with major leaf rosettes and (2001). Herbaceous. fire. Plant height. there are modified versions of the equation Use an asymptotic regression to relate height to diameter. some authors have suggested that plant and the ground level. (2009). References on theory.). For vegetation types with dense and (3) for trees. with a clinometer or laser. measurements are taken preferably on at least the horizontal (as measured with a clinometer) and add the 25 mature individuals per species. the if eye level is below tree base level). Niklas (1994). Westoby (1998). (3) Epiphytes. and may eventually envelope the exceptional branches. the horizontal distance between the observer and References on theory. (Emergent hydrophytes horizontal distance from the tree to the observation point (‘helophytes’) mostly fall into one of the subgroups (d) and. leaf length of two-thirds of the largest leaf as the cut-off point whole-plant fecundity. measure ~25 undergrowth that makes the measurement of Hmax individuals that cover the entire range of their height and diameter. difficult. (1) a telescopic stick with decimetre marks.

g. bud banks and below-ground storage organs many grasses and sedges). certain spp.g.g.5 Clonality.g. underground stems that serve as storage organs and bear either scale or foliage leaves. onion Klimešová 2005). a sharp (4) corms – vertically oriented. occurring after injury and (mostly aquatic plants. structures. For above-ground clonal structures. after a fire that kills the shrub’s aerial canopy. Jerusalem artichoke (Helianthus tuberosus)).g. and bryophytes). can be a very important determinant of recovery and persistence (2) tubers and turions – conspicuously thickened. dig up a minimum of five healthy-looking plants (6) suckers – shoots developed from adventitious buds (Appendix 1). fleshy scale-leaves that act as bank’ concept). that can and are at its disposal for branching. Cardamine If a plant species has clonal growth (Categories B or C above in pratensis. replacement of shoots.g. tulip (Tulipa). that arise after the injury. axillary or what would otherwise be flower buds. produced by secondary growth discussion). potato Solanum tuberosum. aspen partial excavation may give sufficient evidence for classification. In some cases (large and heavy root systems). and following: (8) layering – ordinary vegetative shoots that lie on or bend (1) stolons – specialised. vegetative and/or reproductive shoots grow up plants competitive vigour and the ability to exploit patches rich in from axillary (or sometimes terminal) buds on the key resources (e. (B) clonal organs present above ground. nutrients. For below-ground propagation (e.6).176 Australian Journal of Botany N. which (1) rhizomes – more or less horizontal. clonal if at least one plant clearly has one of the clonal organs (7) lignotuber – a massive. Clonal behaviour may rhizome. fire season).)). or by adventitious bud growth on leaves (e. based on Klimeš and to multiply it vegetatively (e. Viola. Remove the soil and dead plant parts before root connection between them and the parent is severed counting buds or classifying the organs.g. The species is considered or dies.g. and following categories: (3) simple fragmentation of the vegetative plant body (1) regenerative clonal growth. analogous vegetative the present Section). bracken fern (Pteridium)). ground units) and expanding horizontally. including the normally not resulting in clonal multiplication. including the with resprouting from a lignotuber. apparently lie outside the ‘bud concentrically nested. there produce adventitious roots and stems whose axillary bud growth and nodal rooting continue apical growth. thereby producing new ‘ramets’ (above. Both the characteristics of the bud bank and storage organs. globosely thickened distinction between these functions is often impossible. the It is best to assess clonality and bud banks near the end of the sucker shoots can become independent plants once the growing season. especially below-ground (Allium)). can be induced by environmental conditions such as high . dry season. after which be an effective means of short-distance migration under decline and decay of the portion proximal to the circumstances of poor seed dispersal or seedling recruitment. adventitious buds on the lignotuber grow out (A) clonal organs absent. as (C) clonal organs present below ground. clonally generated Clonality also gives a plant the ability to form a bud bank. ones. Clonal organs. which individuals. light).g. (Fragaria vesca). water. also serve as storage and perennating organs. rooting bulblets produced from (Picea) and hemlock (Tsuga)). normally not multiplying the number of individuals. below-ground stems that bear regeneration in some plants. below- after environmental disturbances.g. observe a minimum of (5) tuberous roots – thickened roots that serve primarily for five plants that are far enough apart to be unlikely to be storage but can form adventitious buds that permit clonal interconnected. which are an important means of (3) bulbs – relatively short. becoming independent plants yields ultimately independent plants (e. Iris. below-ground stems. regrowth after severe seasons (winter. often hyper-elongated horizontal down to the ground. propagate the plant (e. Dahlia). non-storage roots (e. 2. ground (e. for Characterisation of the bud bank. of spruce (2) bulbils – deciduous. the whole globose structure serving to the type of clonal growth exhibited by plants determine their perennate the plant and. through growth of axillary ability to recover from disturbances (see Material S3 for a protocol buds within the bulb into daughter bulbs or ‘offsets’. Clonality can give aerial. can be either the plant’s normal mode of multiplication or usually bearing non-photosynthetic scale leaves (e. similar or for vegetative regeneration after injury (adventitious buds organs formed on aquatic plants are termed turions. wild plum (Prunus spp. (Populus tremuloides). strawberry when their connection with the parent is severed (e. most rhizomes can branch. of the ‘root crown’ in many shrubs in fire-prone Categories are then vegetation. axillary or terminal buds on the corm function for perennation and to a How to collect and classify? limited extent for clonal reproduction (e. following: (2) additive (also termed multiplicative) clonal growth. sweet potato (Ipomoea batatas)). Bryophyllum). produced on ordinary. to regenerate the shrub’s canopy (see Section 6. and that are well developed. branch point yields independent. Pérez-Harguindeguy et al. classify it according to one or more of the propagules of bryophytes are termed gemmae. woody expansion just below listed below (see References below in the present Section for the ground surface. saxifrage (Saxifraga flagellaris)).g. and sometimes instead Clonality is the ability of a plant species to reproduce or regenerate bearing photosynthetic leaves that emerge above itself vegetatively. The bud bank consists of all ground stems or rhizomes. blackberry and raspberry (Rubus). functioning as carbohydrate viable axillary and adventitious buds that are present on a plant storage organs and bearing axillary buds.

If a spine Spinescence refers to the degree to which a plant is defended branches. sharp spines >100 mm long. including pruning and is sufficient in some cases. oven-dry and weigh leaves. bulbs.New handbook for measurement of plant traits Australian Journal of Botany 177 nutrient availability. Spine 2. Data on spinescence are preferably Klimešová (2005). In and behaviour of herbivores vary enormously. (5) Intermediate or high density of hard. plant may sting or prickle when hit carelessly. rose-stem prickles). seed reproduction. is often leaves. but rather a trait that reflects the many plants that perennate from rhizomes. plant causes strong pain when hit carelessly. Van Groenendael et al. Special cases or extras in Section 3. than those above. branches. (4) Intermediate or high density of hard. In other words. tuberous roots and have no. but still relatively simple. and ‘tough’ if they cannot be thus bent. cut off all spines. or low density of hard. Spines are ‘soft’ if. Spine density is the against different herbivores. they can be bent easily by pressing sideways with a angles and densities of spines. The number of are modified epidermis or cork (e. spinescence traits can change drastically with the age plant. or weak. e. (3) High density of soft spines. some researchers may decide that experiments with some research questions. characterisations of plant spinescence by measuring spines is Biomass allocation to spines is also an important parameter for sufficient. modified twigs or branches. should also be recorded because branches can Because spinescence is clearly involved in anti-herbivore increase significantly the dangerousness of spines to herbivores. so the degree of Box 3. plant is not necessarily representative of the potential range of in which case it may not be possible to distinguish between spinescence in the whole species (e. as with considered an innate plant trait.Van can either be measured as a quantitative trait or reduced to a Groenendael et al. Thorns are sharp. sizes. but not impart strong pain. Different types. plant causes actual pain when hit carelessly. Cut a standard length herbivore defences. thorns and prickles may act finger. as many do. Weiher et al. tubers or actual herbivore pressure and investment in defence by plants. can also induce increased levels of spinescence. Although in many cases. depending on individuals. thorns and prickles can sometimes play additional roles such. modified branch. age and pruning history). mitigating damage from herbivores. Spines are sharp. the spinescence of an individual Clonal growth may fulfil more than one of these functions. in reducing heat or drought stress. and can also be gathered from herbarium specimens or descriptions in the literature. (1997). significance and large datasets: De Kroon and Van Groenendael (1997). Spine width. or area of leaf.g. depending on its susceptibility to (3) necessary clonal growth is indicated when clonality is herbivory. sharp spines >5 mm long. thorns and/or prickles. plant may cause significant wounds when hit carelessly. and species that never have them. sharp spines >20 mm long. (1998). Klimeš et al. including humans. measured from specimens in the field. Prickles and more generalisable across types of spines. In some cases. its length would be to the tip of the longest by spines. of the plant or plant part. Bear in mind that the size. (4) unknown or not evident.g. in which case it may be recorded as Spines. Spines. categorical trait. meaning that some plants invest in these proposed in Box 3. (2) Low or very local density of soft spines <5 mm long. measured at the base of the spine. Klimešová and Klimeš (2007). mature. and serves to promote the spread of the addition. some members of Acacia them. (1997). (1997). Other types of damage. thorns and prickles – summarised below as ‘spines’ – More on methods: Böhm (1979). the following Ratio of spine length to leaf length can also be a useful character two separate issues are critical in considering spinescence: because it gives an idea of how protected the lamina is by the spine (1) the effectiveness of physical defences in preventing or closest to it. spinescence sometimes cannot be required for the year-to-year survival of the plant. thorns and prickles can be an induced response categorical estimate of spinescence by using the classification to herbivory. if any.16). intermediate density of spines of intermediate hardness. by offering whole of stem or branch. defences only when they have already been browsed by Finally. These quantitative trait measurements can be converted into a Spines. For this reason. shoots (with and without spines) to different animals and shoot and spines separately and estimate fractional allocation as recording how much biomass is consumed per unit time (see the ratio of spine dry weight to shoot dry weight. especially when they densely cover organs.6 Spinescence length is measured from the base of the spine to its tip. leaf parts or stipules. although there are species that always have spines. (1997). are necessary. the functional nature of clonal growth may and Prosopis show a striking range of spine lengths within the be simply same species. plant is dangerous to careless large mammals. number of spines per unit length of twig or branch. which examine the effectiveness of anti. sharp spines >5 mm long. Categorical estimates of spinescence (1) No spines. Klimeš and qualitative. (6) Intermediate or high density of hard. especially against vertebrate herbivores. structure fire. Knevel How to measure? et al. and (2) the cost to the Spine strength or toughness. defence. . References on theory. they also occur sometimes on more useful for assessing effectiveness against herbivores fruits. Klimeš and Klimešová (2005). when plant in producing these defences. to simply record the presence or absence of spines herbivores. (2003). Its estimation is more work-intensive actual herbivores. Klimeš et al.g.

abscission. be taken to select shoots that have not lost leaves or parts of Staver et al. Declining function of sapwood with age is one reason why branching architecture of a plant (a branch that reaches the LA : SA generally increases when one moves from larger (older) outer part of the canopy). which is the distance from the and interpreting this trait (see Point 3 of Special cases or extras in starting point to the tip of its longest-living terminal and (2) the the present Section). Grubb (1992). . called apical dominance index (ADI). is obtained by dividing This approach maximises the likelihood that all the sapwood in the number of ramifications by the total length of the branch in the branch is still functional. Unfortunately. Like photosynthetic rate) and mechanical strength. and ensuring that. Branching architecture is also variable depending on The ratio leaf area : sapwood area (LA : SA) depends strongly the age and life history of the plant (see Section 1. plant vigour or disease and even water What and how to collect stress. of making meaningful comparisons across species. Measurement of branching architecture. access to light. Numbers indicate the plant to continue growing. number of ramifications per meter of branch). there remain enough for Fig.8 Leaf area : sapwood area ratio Although there are complicated and elegant methods for The amount of leaf area a species produces per unit cross-section evaluating branching architecture. When selecting the most meaningful measurement/s of spinescence. herbivory or early senescence and More on methods: Fisher (1986). Note necessary. and within trees along a branch from trunk To assure measuring a branch that best represents the to tip. The value of ADI can vary between zero (no branching) to peak of the growing season when leaf area is highest. An indicator of branching architecture. Strauss and Agrawal (1999). significance and large datasets: Horn to the efforts to measure maximum photosynthetic rate as a way (1971). We recommend sampling at the metres. References on theory. significance and large datasets: Milton 3 2 4 (1991). Archibald and Bond (2003). we each ramification point. Gowda and Palo (2003). LA : SA should be at a maximum for the year. from To make meaningful comparisons among species. leaves to mechanical damage. Olff et al. work backwards from a terminal. expressed in mm2 mm–2) such as the one described below is often sufficient for is crucial for both water transport (with related effects on understanding the adaptive significance of this trait. towards smaller branches. in which terminal bud scars allow their age to be determined). move towards the tip. variation among populations of a given species along moisture gradients. number of ramification points that lead to living branches. Pisani and Distel (1998). a simple characterisation of sapwood (the inverse of Huber value. Agrawal and Fishbein (2006). this is similar References on theory. This means sampling terminal shoots either of a living branch coming from a ramification point. size and distribution can be determined only with reference to a particular kind of herbivore. fire history. primarily by making feeding less efficient. (1999). if herbivores do remove growing tips. pioneer stages of secondary succession. and may vary among species.7 Branching architecture starting point Branching architecture refers to how intensively a plant branches trunk g th (number of living ramifications per unit of stem length).1 for on leaf phenology. All this The base of this branch will be the starting point for measuring should be considered when designing a sampling methodology (1) the total length of the branch. time. as in a fire-prone savannah or a forest undergoing the that dead branches are not considered in the index. wet and dry seasons. denying access by herbivores to plant organs.178 Australian Journal of Botany N. Pérez-Harguindeguy et al. difficult to measure. always consider what herbivores are relevant. 1. (2011). Pickett and Kempf (1980). 1 2. protection provided by spine mass. where species that utilise low light tend to be more branched for a given height than are species that utilise only bright light. Highly le n ch branched plants can be better defended against vertebrate b ra n herbivores. spinescence. there is variation between recommendations related to variable traits). Cooper et al. Hanley and Lamont (2002). (2003). Cooper and Ginnett (1998). 2. can be leafless at its base but bears secondary branches that have leaves. 5 6 Rebollo et al. Branching architecture can also be adaptive in forest systems. always following recommend sampling terminal. Care should Enquist (2002). At this >100 m–1 (extremely ramified). branching architecture is a plastic trait that can differ within a species on the basis of browsing history. or of a certain age (1–3 years) (for shoots graphic explanation). less branched plants ramification points to be considered for the calculation of the apical can be adapted to environments where growing tall quickly is dominance index (i. sun-exposed shoots from the the most important branch (the main branch is often the thickest outer canopy. Furthermore. 1 for a certain standard length. (2002). see Fig. ontogenetic trajectories for How to measure? given individuals. Conversely.e. the age-related decline leaf-bearing branch until reaching the first branch that is now in sapwood function is not always well understood. Gowda and Raffaele (2004).

(3) In ring-porous trees. sometimes within a measured for plants of similar mass. synonymous to root. McDowell et al. such as eosin. Section). seedlings showing however. Preston and Ackerly (2003). the measuring the dye-stained area. Cornwell et al. quantified by placing the cut end of the shoot into a fairly Care should be taken to harvest all the roots (see Section 5). cutting a cross. root allocation measurements).1 for free software). coarse roots. (2002).2). (2007). we designed irrigation systems. and whether this is appropriate or not. For these species. Chapin (1999). Buckley and (1) Storage organs and root fractioning. and after 10–20 min. The RMF is preferable to the (>100–200 mM sodium chloride. Maseda and Fernández (2006). when all the season’s newly produced leaves rate. feature high concentrations of salt follow this convention herein. because root leaves (see Section 3. RMR). Grime (1979). Lower allocation to roots may well be compensated by remain attached. rhizomes (for grasses). (1991). mass ratio. However. care must be taken to identify the parts of the stem plastic responses to low light typically decrease their RMF. and allowing the despite the difficulty of separating roots from soil. Notably. similar methods can be applied. significance and and the final leaf area for the season is attained. low. NaCl). RMF typically decreases with increasing nitrogen availability. Because cambial growth in many trees gradients are potentially confounded by failure to account for continues well after spring flush of bud growth is completed allometry and size (see References on theory. Note that some reports of differences in RMF across resource (2) Seasonal changes. from whole plant to just terminal branches (and compared. however. Aerts et al. sometimes RMF includes section of the stem a few centimetres above its cut end and only a subset of all below-ground tissues. Maherali and DeLucia (2001). allocate a greater fraction of new biomass to roots and References on theory. most precisely measured with digital micrographs and image. (2003). Poorter et al.New handbook for measurement of plant traits Australian Journal of Botany 179 Measuring often used root : shoot ratio (RSR). researcher should be clear about what is included and what is not. in field studies. significance and large datasets: Chiba (1991). point is measured by the same method as the area of individual Some patterns can be apparently contradictory. and those with poorly synonymously. Eamus and Prior (2001). the LA : SA large databases below in the present Section). including mycorrhizae!). Aerts and expressed as the root-mass fraction (RMF. and can be immediately interpreted and scales. Litton et al. particular studies can References on meta-analysis: Mencuccini (2003). length or surface area. tap roots 2. strong solution of a dye. Elberse and than in shoots. allometries can be used to estimate the sapwood (and its decline with sapwood age) can be RMF for plants of a given size. of plant dry mass in roots. In advantage when resources are plentiful. approach. apparently to survive periods of low dye-transport experiment (see Point 3 below in the present water and nutrient supply in competition with surrounding trees. crowns. Veneklaas and Poorter (1998). identically calculated as the proportion (2012). fast-growing species adapted to nutrient-rich habitats showed higher allocation to roots than did slow-growing (1) For herbaceous species. heartwood and pith from the area measured. Similarly. Allocation and distribution are often used poorly drained soils in arid climates. fine roots. theory include everything that is plant-developed (so not (2006). in such a case. species from nutrient-poor sites. including coastal ones.1) and by higher uptake abscission of older leaves has occurred. the conductivity of harvested of a range of mass. It can be quantified with a tend to have higher RMF. subdivide specific fractions for specific purposes (i. Addington et al. because the RFM is bounded Leaf area : sapwood area ratio can be measured at different between 0 and 1.9 Root-mass fraction (in trees)) to evaluate the relative proportions of each in Theory predicts that plants from nutrient-poor sites should relation to each other and/or to above-ground biomass. However.e. including those that take an allometric phloem. Note that a true allocation measurement requires quantifying turnover rates as well as 2. rate per allocation to root mass. being an a calliper should work for most species in most situations. that can conduct water. Sack et al. but (for evergreens) before the seasonal higher specific root length (see Section 5. particularly foliage to transpire in air. Sapwood area at the collection point is allocation can allow both greater foraging below ground. (2007). which would be an advantage especially when resources are analysis software (see Section 3. Total leaf area of leaves distal to the collection can be highly plastic across light. the effective conductivity of xylem The RMF can best be used for comparative purposes if drops precipitously in older sapwood. Special cases or extras Mäkelä and Vanninen (2001). Only salt-resistant . fine roots. those with carried out. nutrient and water supplies. Wright et al. if plants are very few annual rings. (2006). this distinction may not be as clear as whereas plants adapted to chronic deep shade in rainforests it is within most woody species. namely. experimental studies. Alternatively. References on theory. significance and large databases: Evans maintain a higher proportional distribution of biomass in roots (1972). Reich (2002). and also greater competition below ground. ratio is best measured as late in the growing season as RMF does not directly translate to a high soil resource-uptake possible. Additionally. RMF should in Roberts (2006). which is labour-intensive and rarely Many areas of the world. However. whereas the RSR is unconstrained and can vary this should be taken into consideration when scaling up from a tiny to a very large number. Distribution of biomass to roots can be simply Berendse (1993). In reviews of measuring sapwood area.10 Salt resistance standing distributions. care should be taken to exclude bark. other studies have reported that for field Special cases or extras plants.

at intervals of 2 weeks (1) Succulents and halophytes. . The Na+ and K+ concentrations of each measuring soil salt concentrations (as described under sample are to be determined in the laboratory by a standard assay. Plants specialised for inhabiting saline K+ assay method.12. but not for 5 days after are halophytes and occur only in saline environments. Many salt-tolerant succulents or more during the growing season. elevation absorption spectrometry (AAS). indicating salt crystals in dry areas. research into novel approaches and protocols for testing salt Salt compartmentalisation. The salt-resistance traits tolerant succulents are actually also CAM plants). these are accompanied by data on species distribution in saline These glands are often visible (especially under a hand lens) as areas. are termed halophytes. dicotyledonous species. This property Na+. Some species excrete salt from their roots.g. which requires only a simple. involving different values per sampling date. Calculate. Experimental on the surface of the gland. we give several traits and measurements that special glands or bladders on the (usually lower) surfaces of their together help identify a species as salt-resistant. some salt- secondary metabolites) in the cytosol. e. Salt-excreting species eject NaCl through salt resistance. physiological. either actively excrete excess salt or can accumulate NaCl in cell Strong salt-related succulence is found almost exclusively in vacuoles. Leaf samples are to be ground and duration of daily marine inundation (if any) in salt in an equal mass of water. Hopefully. Roots of many salt-resistant Qualitative evidence for this can be a combination of juiciness plant species (particularly monocots) can discriminate against and noticeably salty taste when chewing the tissue. we suggest sampling leaves and soil on at least three different days. Salicornia selectivity for K+ over Na+ increases the K+ : Na+ ratio in the spp. therefore. focusing on three common strategies. previous classifications of stems. by accumulating compatible solutes (including metabolism (CAM) succulents (see Section 3. which can distinguish them from crassulacean acid the tissues. Selective root cation uptake above. The latter (‘salt. one may check for screening multiple species. which exhibit strategies to reduce or avoid damaging vacuum filtration. or white patches on the soil becomes water-saturated and then extract the liquid by suction or surface. species cannot avoid significant NaCl uptake. including Special cases and extras precipitation and evapotranspiration. Succulent green stems can be treated and measured as if mechanisms by which plants deal with excess environmental they were leaves (see Special cases or extras in Section 3.14). necessary. and atomic salt-related habitat descriptors are also relevant.180 Australian Journal of Botany N. testing of plant survival and growth under saline conditions is will confirm this. NaCl. except special be detected by the Na assay on leaf or stem extracts noted above. we simplify more extensive. Many salt-resistant species possess dunes. Therefore. within the present Popular and convenient methods include atomic emission Section) to accompany trait measurements. A mean S halophytes. Selective root cation uptake. Na+ and K+ assays can be performed either on effects of excess salt in their tissues. Thus. Note that salt excretions on shoots or roots and do not allow the clear separation of more or less salt-resistant will wash off during wet weather. Salt compartmentalisation is resistance more efficiently and comprehensively. on licking one of these. the K : Na Among members of the at least 139 plant families that include selectivity (S) as S = ([K+]/[Na+])plant / ([K+]/[Na+])soil. traits. For soil. This has made some halophytes popular as human food. biochemical adaptations that are not covered here. Elytrigia juncea on beach of drought-tolerant species.g. structural and/or phenological sampling dates. e. However. soils. by taking the average of these over all biochemical. A salty taste. Pérez-Harguindeguy et al. many salt-resistant parameters (succulence (mm) = Lth  LWC) (see Section 3. particularly heavy or prolonged rain.g. during soil sampling. and often restricted to these. are able to maintain viable the water phase. to positively classify a species as salt-sensitive small. or after evaporating it.3). depending on the Na+ and populations in such areas. indicated by clear succulence of the leaves or photosynthetic Here. and a soil sample from the main actual salinity of the plants’ soil. which would by no means be quick and easy for Although this is more difficult to observe. also called flame photometry. rather than a single recipe for assessing Salt excretion. We. These plants can Values >800–1000 mm indicate significant succulence. or would be revealed very easily by measuring the electrical conductivity of such extracts (see Electrolyte leakage in Section 3. this text will stimulate period. so are best sought after a dry species from true halophytes. so as to prevent toxicity to the cytosol. which is then extracted from the marshes or on beaches. irregularly shaped white spots that are excreted salt crystals would be problematic from these traits alone. Several other spectrometry (EAS). widely available What and how to measure? conductivity meter (NaCl in solution gives a high conductivity). for each plant and associated soil sample. e. add water to a dry soil until it tide mark visible as a litter belt. especially if photosynthetic organs (usually leaves. the traits described below allow similar salt excretions on the surfaces of any roots uncovered a qualitative rather than quantitative assessment of salt resistance. with many characteristics can be somewhat succulent. Collect leaves from five expression of the traits described above can depend on the separate plants (Appendix 1). and may be quantified as the product of these by their roots (NaCl ‘excluders’). although certain salt-tolerant monocots tolerant’) species are often succulent. Some salt-resistant Succulence leads to high leaf water content (LWC) and leaf plant species can limit the uptake of potentially damaging Na+ thickness (Lth). However.1). while maintaining uptake of essential potassium (K+). but in some cases stems). Salt-tolerant succulents show a high NaCl level in their biochemical mechanisms to reduce salt stress or damage in leaves. cytosol compared with that in the rooting medium. species. This could detailed below fall into the foregoing categories. suggest fine-root zone below each. evolution has yielded multiple solutions to the value for a species is calculated from the mean of all replicate S problem of excess salt in the environment. Because these ratios may vary with several environmental factors. and location relative to the high homogenate by filtration.

strategy with respect to productivity as related to environmental or the total area of leaves. under controlled laboratory conditions or in the field. in case of herbaceous plants. a variable closely RGR ¼ ðlnM 2  lnM 1 Þ=ðt2  t1 Þ: related to the daily rate of photosynthesis per unit LA. the rate of increase in plant biomass per unit LA. several individuals (~10–15 per treatment). each harvest should include a representative sample of the total population studied. Otherwise. Breckle (2002). also known In the case of a well balanced design where plants are paired. the decrease in seed mass between t1 and the different plant parts are oven-dried for at least 48 h at 70C and t2 can therefore be added to M1 (excluding seed mass itself weighed. or they can be measured mass will cause an overestimation of growth.11 Relative growth rate and its components factorised into its components then. However. the average RGR is calculated as morphology (see Section 3. yielding plant fraction. FAO (1999). LA is measured (for details. grown either plant before the averaging. Breckle (2002). reduced by growing a larger number of plants and selecting a where A1 and A2 represent the LA at t1 and t2. probably simplest to calculate RGR and its growth parameters for each pair of plants. The petioles can either may draw on seed reserves for a long time after germination. at least a subset of the materials being studied. Flowers or additionally. gently washing away the soil (see details on procedure under Section 5). The actual The average unit leaf rate (ULR) over a given period is number of plants to be harvested for a reliable estimate increases with the variability in the population. be included in the stem fraction (reflecting support. including roots. RGR cannot be 2. significance and large datasets: Destructive harvests provide a wealth of information. harvest intervals should be chosen follows: such that plants have less than doubled mass during that interval. by their nature. Expressed in this way. (1977). by non-destructively More on methods: Jennings (1976). whereas exclusion of the seed they belong morphologically). the average RGR over Ideally. in the case of juvenile individuals of slow-growing woody Similarly. separately. Growth analysis requires the destructive doing so. In the case of more than two harvests. RGR is the (exponential) increase case. Zhu (2001). stem and root mass as well as LA. good insight into the components underlying growth variation can be obtained in a relatively simple way. Vendramini et al. plants can be plants. Yeo (1983). . Rozema et al. the whole volume of stems Relative growth rate (RGR) is a prominent indicator of plant (and branches) is determined in woody species (see Section 4. a separate sample of leaves (~20) time interval. stem (support and transport) and roots (water (1) Confounding effect of seed size. growth can be followed non-destructively for and Colmer (2008).1). as follows: one plant from each category at each harvest. along with the number in size relative to the size of the plant present at the start of a given of leaves. By harvesting average of the values from the 1st and 2nd harvest. To estimate LA. a more accurate impression of RGR can be obtained. and repeated handling may cause growth retardation.1). with the number of Average leaf mass fraction (LMF) during that period is the plants per category equal to the number of harvests. as well as storage). and then use the RGR values for each pair to What and how to measure? average over the population. By repeatedly measuring the same individuals. grouped by eye in even-sized categories. In the latter stress and disturbance regimes. Munns et al. leaf length and width are measured. respectively. Maas and Hoffman measuring an aspect of plant size at two or more moments in time. These How to calculate RGR? underlying parameters are related to allocation (leaf-mass From two consecutive harvests at times t1 and t2. to more than 2 months or longer where ML1 and ML2 indicate the leaf mass at t1 and t2. As a rule of thumb. the fraction of plant biomass allocated to leaf). size. Alternatively. one initial and one final harvest should be carried out. Size variability can be ULR ¼ ½ðM 2  M 1 Þ=ðA2  A1 Þ  ½ðlnA2  lnA1 Þ=ðt2  t1 Þ. Individuals average RGR can be derived from the linear-regression slope of ln should be acclimated to the current growth conditions. or combined with the leaf fraction (to which the RGR of the new seedling. are extremely labour-intensive and. it is as net assimilation rate). (2002). see Section 3. The harvest intervals may vary from less than 1 week in the case of fast-growing herbaceous species. (2002). By separate measurement of leaf. Alternatively Vendramini et al. but Flowers et al. priori similarly-sized individuals for the experiment. growth rates can be has to be used to determine the linear-regression slope of leaf compared among species and individuals that differ widely in length  width. including leaves (light interception and carbon (C) uptake). destroy (1985). LMF ¼ ½ðM L1 =M 1 Þ þ ðM L2 =M 2 Þ=2.New handbook for measurement of plant traits Australian Journal of Botany 181 References on theory. Again. make sure to first ln-transform the total mass of each harvest of two or more groups of plant individuals. leaf masses M1 and M2.1) before persistent seeds. respectively. In plant. (1977. For large. (2002). this is the Inclusion of the seed in the total plant mass underestimates preferred option). average specific leaf area (SLA) is calculated as species. from M1 and M2). 1986). At least (mass) over time. Especially tree seedlings and nutrient uptake. Plants are divided into Special cases or extras three functional parts. Ashraf and Harris (2004). discarding the simplest option would be to calculate ULR from each pair of the small and large individuals. SLA ¼ ½ðA1 =M L1 Þ þ ðA2 =M L2 Þ=2: At harvest. RGR is measured on a dry-mass basis for the whole the whole group of plants is calculated from the same equation. the whole root system is excavated and subsequently cleaned. Ideally. and physiology (unit leaf rate.

182 Australian Journal of Botany N. Pérez-Harguindeguy et al.

(2) Ontogenetic drift. As plants change over time, they readjust a standard method for measuring flammability in which the basic
allocation, morphology and leaf physiology. Consequently, architectural arrangement of the measured shoots is preserved
LMF, SLA and ULR may change with plant size, and RGR (see More on methods in the present Section for reference, and
generally decreases over time, the more so in fast-growing Fig. 2 for illustration) This involves a low-tech device in which
species. This does not devalue the use of the RGR, as plant shoots up to 70 cm long are placed, preheated and ignited in a
growth does not necessarily have to be strictly exponential. standard way, then the following measurements are taken:
As long as plant growth is somehow proportional to the plant
size already present, RGR is an appropriate parameter that (1) maximum temperature reached during burning (in C, MT),
encapsulates the average RGR over a given time period. measured with an infrared thermometer from a distance of
However, ontogenetic drift is an important characteristic of 50 cm to the burning shoot;
plant growth, and a higher frequency of harvests may provide (2) burning rate (BR), a value obtained by dividing the length of
better insight into this phenomenon. In comparing species or the sample that was burnt by the burning time (in seconds); it
treatments, it may be an option to compare plants at a given gives an idea of how quickly flames can spread across the
size or size interval, rather than over a given period of time. plant and to what extent the plant is able to carry fire; and
(3) Related to ontogenetic drift, shrubs and trees accumulate (3) burnt biomass percentage (BB), consisting of a visual
increasing amounts of xylem, of which large proportions estimation of the burnt biomass (percentage intervals); the
may die depending on the species. This inert mass would intervals are 1 = <1%, 2 = 1–10%, 3 = 11–25%, 4 = 26–50%,
greatly reduce RGR. Previous studies express RGR (‘relative 5 = 51–75% and 6 = 76–100%.
production rate’) on the living parts of large woody plants
For calculating (overall) flammability, the scores on each
by treating the biomass increment over Year 1 as M1 and the
component must be transformed to a proportional scale, with
increment over Year 2 as M2 It could similarly be based on
the value 1 being assigned to a reference value. In the case of
annual diameter (or volume) increments.
BB, the value 1 was assigned to the maximum possible value for
(4) Smooth curves. In the case of frequent (small) harvests, a
this component (i.e. 6), whereas for the other components, the
special technique can be applied, in which polynomial curves
reference value was based on the literature and the results of our
are fitted through the data. This is an art in itself!
experiments. Reference values are: MT = 500C; BR = 1 cm–1.
References on theory, significance and large datasets: Evans Standardised MT, BR and BB scores are then added to obtain a
(1972); Grime and Hunt (1975); Kitajima (1994); Cornelissen compound value of flammability (rounded to two decimals) that
et al. (1996); Walters and Reich (1999); Poorter and Nagel could vary between 0 (no flammability) and ~3 (maximum
(2000); Poorter and Garnier (2007); Rees et al. (2010). flammability).
More on methods: Evans (1972); Causton and Venus (1981); As an alternative to the direct measurement of flammability,
Hunt (1982); Poorter and Lewis (1986); Poorter and Welschen an estimation may be obtained by measuring several plant
(1993); Cornelissen et al. (1996); Rees et al. (2010). attributes that are known to influence plant flammability.
Five classes are defined for each of the attributes described
2.12 Plant flammability below (stem and twig water content, canopy architecture,
Flammability-enhancing traits are important contributors to surface : volume ratios, standing litter, volatile oils, waxes and
fire regimes in (periodically) dry regions and therefore they resins). Flammability is subsequently calculated as the average
have important ecological impacts (particularly on ecosystem (rounded to one decimal) of the class scores for each of the
dynamics), as well as socioeconomic and climatic consequences. following individual attributes (see Table 1 for ranges of values of
The intrinsic flammability of a plant depends on both its traits each attribute within each class). We strongly recommend testing
while alive and the effects of its leaves, branches, and stems after and calibrating this estimation against direct measurements of
those organs’ deaths. The flammability of those organs (either flammability as described above, or against direct measurements
living or dead) depends on (1) the type or quality of the tissue, and of ignitability and combustibility as described under Special
(2) the architecture and structure of the plant and its organs (which cases or extras in the present Section.
is mainly related to heat conductivity). (1) Water content of branches, twigs and leaves. Flammability
Note that the flammability of a given species can be overridden is expected to be greater in species with higher leaf dry-matter
by the flammability of the entire plant community (e.g. amount content (see Section 3.3 for protocol) and higher twig dry-
of litter, community structure and continuity, organic matter matter content (see Section 4.2 for protocol) and it is probably
content of the soil) and by the particular climatic conditions also a function of drying rate (here, represented inversely by
(e.g. after a long very dry period, many plants would burn quite drying time from saturation to dry equilibrium).
independently of their flammability). (2) Canopy architecture. Plants with complex architecture, i.e.
extensive branching, tend to spread fire easily. The degree
How to define and assess? (number of orders) of ramification (branching) is used here as
Flammability (broadly defined as the propensity to burn) is a a close predictor of canopy architectural complexity, and
compound plant functional trait. Its components vary among ranges from zero (no branches) to 5 (four or more orders of
authors and disciplines. Most studies are based on flammability ramification) (see Section 2.7).
measurements of small plant fragments in chambers in the (3) Surface-to-volume ratios. Smaller twigs (i.e. twigs of
laboratory. Although this produces highly standardised results, smaller cross-sectional area) and smaller leaves should
it does not scale up well to whole-shoot flammability. We propose have a higher surface-to-volume ratio (and, thus, faster

New handbook for measurement of plant traits Australian Journal of Botany 183

(l )

(b )

(d )
(j ) ( k)
(e) (h)


(f )

Fig. 2. General view of a device for measuring plant flammability in the field (reproduced with permission from Jaureguiberry et al. 2011). (a) Grill, (b) grill
thermometer, (c) temperature gauge, (d) security valve, (e) connection to gas cylinder, (f) removable legs, (g) blowtorch valve, (h) blowtorch, (i) burners,
(j) ventilation holes, (k) barrel, (l) removable wind protection and (m) gas cylinder. See Jaureguiberry et al. (2011) for technical details.

Table 1. Plant flammability traits
Classes for component traits for estimating plant flammability. Flammability itself is calculated as the average class value (rounded to 1 decimal) over all
component traits. Flammability increases from 1 to 5 For this calculation, twig drying time (which is probably closely negatively linked with twig dry-matter
content, TDMC; see Section 4.2) is optional

Component trait Flammability class
1 2 3 4 5
Twig dry-matter content (mg g ) <200 200–400 400–600 600–800 >800
Twig drying time (day) 5 4 3 2 1
Leaf dry-matter content (mg g–1) <150 mg 150–300 300–500 500–700 >700
Degree of ramification (branching) No branches Only 1st-order 2 orders of 3 orders of 4 orders of
(number of ramification orders) ramification ramification ramification ramification
Leaf size (lamina area) (mm2) >25 000 2500–25 000 250–2500 25–250 <25
Standing fine litter in driest season None as one Some Substantial More dead than Shoot dies back
litter unit (with dead leaves or live fine mass entirely, standing as
twigs or flaking bark) above ground one litter unit
Volatile oils, waxes and/or resins None Some Substantial Abundant Very abundant

drying rate) and therefore be more flammable. Since twig and attached to the plant during the dry season is critical, since
leaf size tend to be correlated in interspecific comparisons, litter tends to have very low water content and thus enhance
according to allometric rules, we use leaf size here to plant flammability. ‘Fine’ litter means litter with diameter or
represent both traits. A complication is that some species thickness less than 6 mm. We define five subjective classes
are leafless during the dry season; however, leaf litter is likely from no ‘fine standing litter’, via ‘substantial fine standing
to still be around in the community and affect flammability litter’ to ‘the entire above-ground shoot died back as one
during the dry season (see Section 3.2). However, for ground standing litter unit’.
fires, substantial accumulation of litter of small leaves may (5) Volatile oils, waxes and resins in various plant parts
pack densely, obstructing oxygen flow and actually strongly contribute to flammability. This is a subjective, categorical
inhibiting fire to spread. trait ranging from ‘none’ to ‘very high concentrations’.
(4) Standing litter. The relative amount of fine dead plant Check for aromatic (or strong, unpleasant) smells as well
material (branches, leaves, inflorescences, bark) still as sticky substances that are released on rubbing, breaking or

184 Australian Journal of Botany N. Pérez-Harguindeguy et al.

cutting various plant parts. Scented flowers or fruits are not Van Wilgen (1996); Schwilk and Ackerly (2001); Gill and Zylstra
diagnostic for this trait. (2005); Scarff and Westoby (2006); Cornwell et al. (2009);
Pausas et al. (2012).
Special cases or extras More on methods: Papió and Trabaud (1990); Stephens et al.
(1994); Valette (1997); Dimitrakopoulos and Panov (2001);
(1) Ignitability indicates how easily a plant ignites (i.e. starts to
Etlinger and Beall (2004); Scarff and Westoby (2006); Van
produce a flame). It can be measured directly by measuring
Altena et al. (2012); for flammability of whole shoots
the time required for a plant part to produce a flame when
described above, see Jaureguiberry et al. (2011).
exposed to a given heat source located at a given distance.
Ignitability experiments are usually performed several times 2.13 Water-flux traits
(e.g. 50), and the different fuels are ranked by taking into
account both the proportion of successful ignitions (ignition Plants play a key role in most hydrological fluxes in terrestrial
frequency) and the time required to produce flames (ignition ecosystems, including the capture of precipitation, retention
delay). Tissues producing flames quickly in most of the trials and spatial (re)distribution of water above and below ground,
are ranked as extremely ignitable, whereas tissues that rarely and water loss through evaporation and transpiration. There
produce flames and/or take a long time to produce them are are considerable differences among species in the extent and
considered of very low ignitability. These experiments are manner in which they influence water fluxes. These species-based
run in the laboratory under controlled conditions (moisture differences affect not only their own growth and survival, but
and temperature) by locating a heat source (e.g. electric also influence key ecosystem processes such as growth and
radiator, epiradiator, open flame) at a given distance (few nutrient cycling through direct and indirect effects of moisture
centimetres) from the sample. The values used to rank species distribution. This section focuses on plant traits that affect
according to ignitability depend on the type and power of the hydrological fluxes external to the plants, and therefore
heat source, on the distance of the heat source to the sample, excludes traits related to the flow of water through plants or
on the shape and size of the samples and on the relative the storage of water in plants (see Sections 3.3 and 4.4, among
humidity of the environment in the days before the test; these others). Differences in the impact on fluxes among species are
experimental conditions should be kept constant for all trials mainly due to differences in plant architecture, morphology and
and samples. We propose using an open flame at 420C, surface features, which can all be quantified. The effects of litter
placing the plant material at 4 cm from the flame. A standard and small plants on surface runoff and infiltration, and the effect of
quantity of 1 g of fresh material is used. roots on below-ground movement of water through preferential
(2) Plant tissue heat conductivity (combustibility) can be pathways and altered soil hydraulics, can also be important;
assessed by the heat content (calorific value, kJ g–1), however, they are not further discussed here.
which is a comprehensive measure of the potential thermal A large fraction of incident precipitation hits plant surfaces
energy that can be released during the burning of the fuel. before reaching the ground. The fate of this water can be (1) free
It is measured with an adiabatic bomb calorimeter using fuel throughfall, (2) retention followed by evaporation or uptake into
pellets of ~1 g, and the relative humidity of the environment the leaf, (3) release as throughfall, as a consequence of drainage
in the days before the test should be standardised as from saturated surfaces or (4) drainage via twigs and stems
well. Evidence shows that heat content varies relatively (stemflow). Precipitation reaches the ground as throughfall or
little among species and is only a modest contributor to stemflow. Rainfall interception regulates the amount of water
interspecific variation in flammability. reaching the soil (and plant roots) and may ensure a less erosive
(3) Combustibility and structural variables. In relation to the water supply during and after heavy showers. Rainfall
surface area-to-volume ratio, other structural variables have interception is influenced by the density of the plant crown
been used to characterise the combustibility, especially and its ability to retain water on its surfaces. Water retention
the proportion of biomass of different fuel classes (size on plant surfaces is greatest in low-intensity rainfall in the absence
distribution). Typically, the fuel classes used are the of wind. As vegetation surfaces approach saturation, further
biomass fractions of (1) foliage, (2) live fine woody fuel interception of precipitation leads to drainage of excess water
(<6-mm diameter; sometimes subdivided in <2.5 and (but see fog interception under Special cases and extras in the
2.5–6 mm), (3) dead fine woody fuel (<6 mm), and (4) present Section). Plant traits that determine the actual water
coarse woody fuel (6–25, 26–75, >75 mm). The summed retention on plant surfaces in a given environment are surface
proportion of live and dead fine fuels (foliage and woody of angle and ‘wettability’. Hydrophobic surfaces have sharp contact
<6 mm) may be the best correlate of overall surface area-to- angles and droplets tend to remain separate, whereas hydrophilic
volume ratio. surfaces have small contact angles and water spreads over the
(4) Fuel bulk density (= fuel weight/fuel volume) and canopy surface as a film. The surface traits that most affect wettability are
compactness (ratio of fuel volume to canopy volume) have cuticular waxes and trichomes.
also been used to characterise heat conductivity, mainly at
the population and community levels. Furthermore, high How to assess
litter fall and low decomposition rate will increase the
(1) Gap fraction. The openness or gap fraction of a plant’s
combustibility of the community.
crown is an important determinant of the free throughfall
References on theory, significance and large datasets: Mutch fraction (p) (as well as its light interception). For each species,
(1970); Rothermel (1972); Bond and Midgley (1995); Bond and we recommend taking measurements of individual plants,

a hose leads the captured water into a collector. a flexible gutter- shaped form (such as a hose cut in half) is fixed around the (5) Droplet retention ability. the upper necessarily equal to the gap fraction measured. If digital pictures have enough contrast. where rn is the distance large enough to saturate the crown. Stemflow can be quantified by attaching collars rainfall. neighbour foliage below the crown. The most realistic estimates are obtained using natural large as a result of clustering of foliage and channelling or simulated precipitation with plants in the field. type leaves. It can be useful to take these patterns into account by and pesticide spray literature. p is not position on the plant or leaf age. How easily a leaf can get wet is from wet leaves. For small-scale applications. relative to foliage cover (dark) within the crown outline. See Special cases measurements. leaf wetness and interception by accelerating drainage (4) Leaf wettability. It is useful to of water because stemflow concentrates the water running off measure droplet-retention ability on the same leaves as those large areas of foliage. particularly isobilateral and needle- and extras in the present Section for direct measurement of p. measurements on both sides of the leaf are or spirals around plant stems that channel the water running recommended. projected area. For most leaves. Variability of throughfall under plants is very field. This parameter can be because part of rainfall striking vegetation may not estimated at the whole-plant level or for plant parts such as be retained because of the force of the impact or the leaves and stems. The parameter p can be estimated stems and the extent of branching. How well water ‘sticks’ to a leaf stem and sealed with silicone or other sealant. From graphically as the slope of the regression line that describes the perspective of plant traits. because rain (usually adaxial) surface is the logical surface for wettability drops do not always fall strictly vertically. For others. it is important to do comparisons under similar rainfall conditions. to which different plant species channel precipitation to their stems. the inclination angles of (1) Free throughfall (p). leaf surface and measuring the angle of inclination of the leaf Collars and collectors need to be able to handle large volumes at which the droplet first begins to move. stemflow is best expressed as a the relationship between throughfall and rainfall. angle is its calculation based on measurement of the contact imaging software can help calculate the fraction of sky (light) area between droplet and leaf.g. as precipitation events that are of insufficient size and follows: intensity to cause drainage from these plants. Rainfall retention sampling along radii of the plant. The lens has to be held neatly measurement of the angle. The procedure involves (1) weighing the movement of crowns. Gap fraction can be The microscope can be fitted with a goniometer for direct estimated photographically. troughs or large sheets. using percentage of the volume of rain falling on the plant. in a situation of negligible between the nth collector and the centre of the plant. At the bottom can be measured by placing a droplet of water on a horizontal of the gutter. Measurement of drip-tip length involves a A droplet of standard volume (2–5 mL) is pipetted onto the decision about the position of the base of the drip tip. rainfall simulators may be used for comparative studies. In most cases. An alternative to the measurement of the contact but not much more. which is unable to droplets and the leaf surface. or images can be obtained with a horizontally below the canopy and capture the entire crown camera. Usually. If precipitation is recorded continuously. Choose time or facilities are limited. the capacity for water storage on stems. For a quantitative measure of the extent used for determination of leaf wettability. which is seldom reached in the dataloggers. because these influence e. down the stems into a collector. or to use rainfall simulators. respectively. They typically include standard commercial amount of water retained can be expressed in different ways. thus reducing the duration of surfaces have greater contact angles and are more spherical. If The sampling strategy depends on the research aims. It is . Droplets on water-repellent retain the accumulating water. observations made stemflow ½% ¼ stemflow ½L=ðrainfall ½mm immediately after commencement of large precipitation  crown projected area ½m2 Þ  100%: events can be used as well. Wetting can be achieved by immersion or Continuous measurements of precipitation utilise devices by simulated rainfall or fog. fog or mist. as well as leaves that are often exposed to inclined (2) Stemflow. leaf wetness. Water is channelled towards the long and determined by measuring contact angles between water narrow tip at the low end of a hanging leaf. Designs towards the centre of the plant or towards the outer parts of the for rainfall or fog simulators can be found in the soil erosion crown. This provides estimates of retention where (2) Drip tips. the gap fraction will be a leaves randomly or select in a standardised way according to leaf reasonable predictor of free throughfall. free throughfall will be slightly overestimated (3) Water retention on plant surfaces. among other things. Immersion will tend to give such as tipping-bucket mechanisms that produce output for maximum water retention. Methods for quantification of plant (or part) without surface water. Stemflow varies with Special cases or extras plant size and architecture. Representative (equal-area) can also be estimated as total interception (rainfall minus sampling of concentric parts of the crown is achieved by throughfall and stemflow) of a discrete rain event that is just following the rule rn = Hn  r1. rain gauges (pluviometers or pluviographs) or custom- including water per unit plant surface area or per crown made collectors such as funnels. These are morphological features that influence weighing is impractical or impossible. However. The literature. (2) wetting of the plant rainfall and throughfall are described in the hydrological (part) and (3) weighing the plant (part) with wet surfaces.New handbook for measurement of plant traits Australian Journal of Botany 185 with minimal canopy overlap with neighbours and minimal leaf surface and observed from the side with a microscope. evaporation.

We then recommend additional measurement of the two Narrow structural elements with thinner boundary layers component variables. but not always. seedlings. recommended that this is established by drawing the tip of a Specific leaf area is a function of leaf dry-matter content (see normally tapering (acute or obtuse angled) tip. avoid leaves with obvious symptoms of pathogen or accounted for. In general. Any rachis (stalk-like midrib of a compound leaf) and all veins are considered part of the leaf for a standardised SLA measurement (but 3 Leaf traits see Special cases or extras below in the present Section. In these. across individuals or species) to sample outer canopy leaves References on theory.4). Martorell and Ezcurra (2007). For true shade species (those that never grow in full et al. or with a substantial cover of epiphylls.3). Pérez-Harguindeguy et al. A certain fraction of water on contribute to SLA to different degrees. Water uptake is particularly efficient in influential. Ideally. Its interception can be common. Epiphyte load can be herbivore attack. light-saturated photosynthetic rate and with leaf nitrogen (N) which may lead to shrinkage of the leaves and therefore somewhat concentration. high ratio of crude fibre to protein) are gravity (‘horizontal precipitation’). area in the direction of air flow (e. sections with the leaves still attached and not removing the divided by its oven-dry mass. Domingo et al. (1990). Lth can be equally arid environments. and thus presumably resource-limited. Section 3. Veneklaas sunlight. significance and large datasets: Skinner (also called ‘sun leaves’) from plants growing under relatively et al.g. and probably stemflow (increasing leaves from adult plants (unless the research is focussed on e. slow-growing plants with (4) Fog interception. Fog consists of small water droplets sclerophyllous leaves (with thick epidermal walls and cuticle. deposited on plant surfaces through air flow rather than abundant sclerification. (2005). A simple method to determine rates and amounts of water Oxalis) have a high SLA and low Lth. samples in moist paper and put them in sealed plastic bags. Wherever fluxes).186 Australian Journal of Botany N. In cool-temperate herbaceous water on wet leaves represents ~0. or expanding or senescent leaves). This is more of a problem for such as tannins or lignin. overstorey. the samples (twigs with leaves attached) should be environments tend. Some species that normally grow in deeply shaded. Then. Herwitz (1985). leaf dry-matter content and Lth. soil-nutrient limitations. collect leaves from the least shaded parts found (but Núñez (2007). their interception and retention research questions it is best (giving the fairest comparison properties can be quantified as described above. SLA tends to scale positively with mass-based Plants in the field may be dehydrated to an unknown extent. Brewer and sunlight). Wherever epiphyte load is important. more than to high leaf thickness. common for epiphytes. (1991). Brewer et al. although there can also be end submerged in deionised water. Therefore. are simply 1/SLA).g. for many large epiphytes). up. namely. SLA assessed as mass. can hugely influence water interception photosynthetically active) but fully expanded and hardened of rainfall and fog. SLA and its components are often. Point 3 above in the present Section) and aerial roots. a tree crown) and the dimensions of canopy elements. Puigdefábregas and Pugnaire optimal conditions. SLA is frequently used in growth analysis because it is often positively related to potential RGR Storing and processing across species. (Note that leaf mass per area leaves until just before processing. cross-sectional area of related to each other and to productivity gradients in a simple way. Typically. low SLA of slow-growing species tends to be related to most of this water will have evaporated before it can be taken high leaf dry-matter content. Although datasets. specific leaf mass (SLM) and specific leaf weight (SLW). with the cut those in resource-poor environments. and Lth (see Section 3. e. Trichomes (see as well as SLA. deserts after a rain event) resource-rich leaves. In that . Burd (2007). For species that typically grow in the (1999). (1989). When woody perennials are dominant. in the absence of vegetation that captures it.2-mm depth. David et al. and negatively with leaf longevity and C unreliable measurements of LA (see Special cases or extras investment in quantitatively important secondary compounds below in the present Section). sclerophyllous or temporarily (e. as a covariate. to have a higher SLA than do cut and immediately placed into test tubes or flasks. which in some cases can be species-specific Select the relatively young (presumably more but generally is not. In contrast. Both components can (3) Water absorption by leaves. As a consequence of these traits affecting the rate of fog interception are the surface variations. low SLA is associated with high leaf dry- a net gain because fog would not normally precipitate in the matter content more than with high Lth. water uptake into the leaf can be significant even in non. (1998). on average. for a 3. the and the plant group in question. water retention on the host plant and modifying water g. Meyer (1994). depending on the habitat leaf surfaces may be absorbed by the leaf.1 Specific leaf area discussion on this and on whether petioles should be included in the measurement).g. it should be possible. If this is not feasible. may further increase fog capture. We recommend collecting whole twig Specific leaf area (SLA) is the one-sided area of a fresh leaf. micro-habitats (e. What and how to collect? (5) Epiphyte load. Fog interception can be succulent plants that are common in some seasonally dry particularly important to many epiphytes or to plants from dry subtropical to tropical areas.g. low SLA is associated with low substrates such as rocks and coastal deserts. In areas with severe uptake is by weighing. (LMA). plants with specialised trichomes such as certain bromeliads. are relatively efficient in capturing fog. as percentage cover or as counts (for is strongly affected by light intensity. take leaves from plant parts most exposed to direct More on methods: Aston (1979). wrap the considerable variation in SLA among co-occurring species. The main plant leaf dry-matter content and high Lth. not from those that look light-stressed or bleached). species in permanently soft-textured high-SLA leaves than for low-SLA.

once taken from the oven. whereas the main function of the leaf blade is completely within the scanning area. it is better to store the samples in plastic After the area measurement. Put them therefore in a with SLA values higher than 10–15 m2 kg–1. make the decision that best suits . inclusion of the petiole may reduce the scanned in the laboratory. therefore. Some Measuring authors consider that the petiole is an integral part of the leaf Each leaf (including or excluding petiole. dry in paper bags. In situations where desiccator with silica gel until weighing. Xerophytic and especially succulent leaves replicate. although generally improve the accuracy of the weighing. such as those will take up some moisture from the air. Always calibrate the area meter by using main function of the petiole is the spatial positioning and pieces of known area before measuring leaves and always check hydraulic support of the leaf. but in clearly be considered leaf and background Coloured scans will also allow different sectors. or at (2–6C) are essential to avoid rotting. http:// and if recollecting would be difficult. or else back in the oven the rehydration procedure described above cannot be applied. with two lamps Therefore. Be aware that. UK. see Special cases or extras 24 h. highly resinous leaves) rot very downloadable from the Nucleo DiverSus toolbox. until further separation of leaf and background. e. calibration. with a digital it can be a source of considerable and systematic error when camera.nih. 80C for 48 h (avoid higher temperatures). USB. low temperatures below in the present Section). Including comparing different studies. Cutting leaves into smaller pieces concentration and air humidity. should not be rehydrated for more than 6 h. University of Sheffield. try both moist and dry www. accessed 22 February 2013) and GIMP (from storage simultaneously and use the dry-stored leaves in the case of the GNU Project. When using digital images. A camera mounted on a tripod. or even in certain same-site a ruler or an object of known size in the image allows for size comparisons of species with very different leaf structures. Leafarea (A. preferably within according to the objective of the study. Weighing several tiny leaves as if they were one and storage in sealed. Try to then oven-dry and weigh petioles belonging to a position the leaves as flat as possible in the position that gives the replicate separately from the rest of the leaves from the largest put each leaf sample in the oven bags without any additional moisture.gnu. or separately according to the objectives of the study. on the monitor) that the whole leaf is positioned flat and of the stem.g. possible after collecting and rehydrating. An important issue is whether or not petioles should be included in SLA measurements. Projected area (as in a which the leaf cannot be displayed.g. If storage is to last for more than 24 h. If this process fails. see Box 1) quickly when moist and warm. LA can be measured with transpirational water loss. Askew. calculated SLA drastically. so that they can be measured together for post hoc measurement of other features of interest. If no cool box is available and processing can be found at Prometheus Wiki (see Box 1). is to measure leaf blade and petioles separately. and store the bags in the the samples leaves subjected to dry storage or for ‘soft’ leaves. we Special cases or extras recommend collecting leaves of these species in the morning after a rain event. (1) Petioles. in mildly succulent species). Store the collected samples in a 2013). some species with very small. bromeliads. Measure as soon as (petioles and laminae either separately or in the same envelope. cacti. by running a preliminary check against LAs 50%. The appropriate decision depends on the research question at hand. ImageJ (from the US National Institutes of Health. which will minimise may facilitate flattening. In both cases. rather than for the average of several replicates. thus resembling the function (e.g. ideally at 70C for at least 72 h. using a range of different leaf sizes. whatever the storage or rehydration method used might be. If you are to use a portable light interception and C fixation. e. In all cases.g. thereby facilitating the advantage that many scanners can draw their power via comparisons with other studies. make sure the leaves are not curled-up or overlapping. P. http://www. therefore. they are better stored or. to re-dry. UK) or LI-COR the petiole should not be included in the SLA because the (Lincoln. Freely downloadable programs are Tissues of some xerophytic species (e. moist plastic bags (with or without addition of then dividing the weight by the number of leaves will generally damp paper) for 12 h is an acceptable option. e. they include photograph) can be measured with specialised leaf-area meters petioles in SLA measurements. The fraction of leaf dry LA meter. Therefore. but without squashing them to the extent that the same replicate.New handbook for measurement of plant traits Australian Journal of Botany 187 case. or a few hours after generous watering. make sure that the estimation error is not too high for mass represented by the petiole varies from ~zero to almost your purposes. in the laboratory. the best (albeit more time-consuming) option lighting from different sides and no flash gives the best results. so that A third option is to determine LA with a flatbed scanner (with SLA can be calculated in both ways. Scan in colour mode to obtain we suggest to scan or photograph petioles and the rest of maximal information for the threshold level between what is to the leaves of each replicate in the same image. Although inclusion or not of the Images of leaves can also be electronically captured in the field or petiole may sometimes not be crucial within a single study. always do so for each individual rehydration method. Additional details on image processing in the laboratory. Other authors consider that such as those from Delta-T Devices (Cambridge. If in doubt (e. NE. accessed 22 February rotting of the moist-stored ones. from a laptop).g. When converting SLA into yields approximately ~5% lower values than does the complete LMA values or vice versa. temperatures are high. In general. and and extras below in the present Section) is cut from the stem and because it provides support and a vascular system without gently patted dry before measurement. then determine the Rehydration is preferable for most plants and essential for dry mass. breathe into the bag before closing it to enhance CO2 tissue might get damaged. see Special cases because it is shed at abscission together with the leaf. for more complex analyses including other plant organs. USA). Transform images to the HSV colourspace for a better cool box or fridge (never in a freezer) in the dark. image analysis software. and stored for later processing. Leaves are pressed gently under a glass plate.

and the practical steps in each case. only the Another decision to make in the case of compound leaves is lamina is considered. however.) have very tiny scales closely appressed to fine soft calculation.12). the decision on which measurement to take and petiole. pressed flat in envelopes Juncus) and the ‘branches’ of horsetails (Equisetum) or or in a plant press. The accuracy of the initial measurement of the saturated area younger stems of some rushes and sedges (Eleocharis. whether to measure the SLA of a typical individual leaflet as in the case of petioles (see Point 1 above in the present or that of all leaflets taken together. take the plant desiccate. Be aware than in some plants. Pérez-Harguindeguy et al. bags. excluding the leaf sheath. We recommend taking both measurements. Callitris considering the rachis as part of the leaf for the SLA sp. leaves that tend to curl. In not extremely tall trees. This sometimes poses practical point. To measure this trait. cases. Clearly. the research question. For a rough measurement. As in the case of rachis Section). For ferns. above. flat leaves. (8) Succulent and leafless plants. the choice of measurement depends on which could be accessed with a pruner on an extension pole. such as leaf-rolling in some grass consider exposed leaves halfway the crown length. we recommend taking several measurements for herbarium material and fossils) and an ‘pastry cutter’ portions (of known area) of young but easily measured trait that reflects multiple structural and fully hardened leaves (e. Once they have been placed in plastic provide the best insight. taking care that the same surface is measured. present Section).2) and LDMC (Section 3. However. Sometimes species have a lower SLA sure that your equipment is sensitive enough to adequately because of their thick rachis. The maximum area shrinkage reflects differences could mean taking the top 2 cm of a young twig. Although this internal LA (Section 3. similar green leafless shoots can be treated as leaves too. the rachis can a leaf analogue. because they are shed as a single unit. In such cases. There are. cut the leaf up into smaller parts and . True leaves from some species (e. whereas total LA is related to the total some workers rely on professional climbers. Then oven-dry the leaf. The decision of whether to measure such leaves. In ‘standard’ leaves. however. specify in your publication could be measured following the standard routines. (10) Leaves of tall trees. your study objectives. Measure fresh parenchyma does not always contain chlorophyll and projected LA. For plants whose main (3) LA shrinkage. part that is the functional analogue of a leaf and treat as calculated as 100  (1 – dry area/saturated area). Shrinkage is defined as a percentage. and Lth. This may be Because many of these species occur in a range of difficult or impossible for some needle-like leaves. pressure–volume whole leaf or equivalent (e. Usually. this species. For some species with photosynthetic spines or Shrinkage averages ~20% and can reach 80% for some non-succulent stems (e. For in leaf structure and is correlated with other leaf traits cacti and other succulents. smaller flat pieces. is crucial. are reduced. leaflet or the whole leaf. Several species have of the study.g. often seen as green or (4) Projected v. an alternative could be to however. Ulex.g. hydraulic properties of a leaf. Needle-like leaves are a specific case wrinkling and folding. Calculate shrinkage using the formula (9) Ferns. If leaves are larger than the window where projected and total LAs are different. only collect fronds (fern ‘leaves’) given above. large leaves may be put in a hard-cover folder to avoid (5) Needle-like leaves. we recommend taking a including cuticular conductance. plus the and measure leaves following the exact same protocol as for internal succulent parenchyma. we recommend to clearly specify in the much easier managed by cutting leaves into shorter pieces publication whether the area reported is that of an individual of 5–10 cm.g. because the leaves are generally narrow. a cladode in Opuntia) parameters such as modulus of elasticity and turgor loss whenever possible. Upper-canopy leaves of sun-exposed sided LA. However. in Agave) or ‘ribs’ (in cacti). measure leaf length with a ruler and leaf width with a are similar to those on petioles (see Point 1 above in the calliper and subsequently multiply 2  length  width. LA shrinkage is columnar cactus are often too big to process or even to both a potential problem (causing biases in leaf-area collect. where both light interception and gas exchange outer half of the crown (inner leaves are sometimes older).188 Australian Journal of Botany N. In such cases. the decision largely depends on the objectives depends on the research objectives. the projected LA is smaller than the total one. e. total LA. be more than 10 times heavier that the sum of the leaflets. This step is crucial because some leaves may SLA measurements. If these cannot be easily reached. it has an essential role in the CAM experience very little shrinkage in area (5%). slingshots or amount of photosynthetically active tissue. at the species. Cut leaf into pieces in the cases when the leaf therefore some authors recommend not considering it in is not flat. a whole Agave leaf or a rib of a plant growth form and deciduousness. without the spore-producing sori. and environments. it is important to specify the exact method especially thick leaves may need to be broken into several used in each case. As a default option. Measure projected LA on the dried leaf. you might treat the terminal twiglets as the study. (7) Leaves of grasses and grass-like plants. They are generally and above all.g.g. Projected LA is generally related to light trees should be preferred. we recommend (6) Tiny leaves. whether petioles are included or not. interception. or even roll up. margin of the frond. make (2) Compound leaves. so the metabolism of succulent plants (see also Section 3. although knowledge of both would (11) Very large leaves. Senna aphylla). as well as difficulties.3). LA is brown structures of various shapes at the lower side or measured as a one-sided projected LA. in non. guns. Leaves decrease in size when they photosynthetic organs are not true leaves. collect including epidermis and mesophyll on both sides. Projected LA of the area meter. and indicating this clearly when reporting twigs. leaf dry-matter content (LDMC). you can include the rachis or not.

In the case of a thick central variation in the LA may also be linked to allometric factors (plant vein. remove with a scissor the protruding upper or lower size. (2007). anatomy and architecture. Because leaflessness is an important leaves. There are situations in which taking fresh leaves. In the case environmental nutrient stress and disturbances. Wilson et al. Therefore. Paula size. entire sample leaf analogues (see Succulent and leafless plants in leaves. For storing leaves. and the appropriate decision depends portable scanning devices cannot be transported to or on the research question at hand. (2002). However.1. the leaflet powered in the field site. but include it in the dry-mass measurement. This need to be excluded from certain data analyses. Ideally. (1992. Alternatively. and always report quick and accurate way to compare leaf laminae. where heat stress. Species with alternate. because this allows one to address borer). expression of canopy dominance. this in your publication. Another practical and inexpensive alternative is measured. leaves of both types in proportion to their estimated contribution to total LA of the plant. either the leaflet area or the leaves to the laboratory for scanning is not feasible. to use a digital camera (see Measuring above in the present When analysing total light capture. See Section 3. ribs and petioles. SLA Special cases or extras measurements obtained in this way should not be compared with or combined with those taken on whole (1) Leafless plants. as a result. (1999). collect Section 3. Juneau and Tarasoff heat loss than do broad leaves with the same area. More on methods: Chen and Black (1992). 1999). (2009). In the case of species with two or number of replicates (i. with a punch. tend to have a smaller boundary layer and a more effective Hodgson et al. xerophytic Section 3. (3) Ferns. Westoby et al. References on theory. and placing the fragments in more questions and to compare the results with other studies.g. Vile et al. at least not without a specific calibration.1). considered adaptive in warm. (2012). 2001b). Within climatic zones. and length halfway. determine for compound-leaved species both obtaining leaf fragments of known area (e. Note that this whole LA may be different it overestimates SLA as compared with measurements from the area used to determine SLA. Plants are modular The area of a leaf (also called leaf area. This is measured as the maximum diameter of et al.g. more positively than does the area of the whole leaf to the Niinemets et large-sized leaves. geology.g.1). Díaz et al. method works well for medium. There is also emerging evidence that leaf width contributes (2001a. For the heat balance. close to the higher end of the number of more types of leaves with contrasting shape and anatomy. (13) Low-tech options for the measurement of SLA and LA Measure the individual leaf lamina for species with simple in the field. be aware that these zeros may the leaves in the field. Measure the laminae with or without petiole and rachis.1 LA is a variation in SLA over the leaf. thus overestimating the LA. If you want to rely on subsamples of leaves. the leaflet area and whole LA. leaf number. plants with both rosette and stem leaves. cold additionally a crude estimate of the size of each growth . an envelope for drying (take several punches per replicate. Blonder et al. Castro-Díez et al. (2005). significance and large datasets: Reich (4) Leaf width.1. according because they tend to weigh very little). rather variable within plants and we recommend collecting a large (12) Heterophyllous plants. these functions can be metric for leaf size and is defined as the one-sided or projected partially uncoupled. and measure their area later. Leaves with very stress. For compound-leaved species. expressed in mm2 (see Section 3. drought stress. (2) Heterophyllous plants. and leaves that are not too narrow (e. or divided leaves with narrow lobes. opposite and area of an individual leaf. number of part of the vein and scan the leaves without that removed lateral buds produced) and ecological strategy with respect to part. Garnier and Laurent (1994). (2011). sun-exposed environments. and Pausas (2006).New handbook for measurement of plant traits Australian Journal of Botany 189 measure the cumulative area of all parts. Leaf size is a compromise between 3. One solution in these cases is area is important. or whole LA can be measured. Vendramini et al. Wright et al. Narrow leaves. nutrient stress and high-radiation stress all thick veins or rachis can cast a lateral shade on the LA meter. record LA as zero for leafless species (not alternative is to obtain plastic or paper prints or cut-outs of as a missing value). altitude and latitude. See Section 3.e. whorled leaves frequently co-exist and leaf dry mass or Interspecific variation in LA has been variously related to climatic area multiplied by the number of leaves per node provides variation. (2000). make sure that there is not too large of For the leaf-collecting protocol. and phylogenetic of a thick rachis. and calculate the rachis area as the product of the two. see under Section 3.1). grasses). (2004). This method is a to the objectives of your study (see Section 3. the whole leaf should be Section). Garnier et al. (5) Leaf number per node. Another functional trait. remove the rachis and measure its diameter factors can also play an important role. especially in the case of large leaves with thick veins. see such as e. (2007). which is functionally analogous to a simple leaf. so as to obtain a Measuring representative SLA value of the individual. twig size. of whole leaves. (2002). This is (2012). tend to select for relatively small leaves. LA) is the most common in construction and. Then scan the leaves without rachis but What and how to collect? include the rachis in the dry mass.1. replicates recommended in Appendix 1). Garnier (1999).2 Area of a leaf functional and resource-use efficiency. and is an additional trait of ecological interest related to leaf Niinemets (2001). Poorter et al. Poorter and an imaginary circle that can be fitted anywhere within a leaf. avoiding thick veins. However.

Optimisation theory.3 Leaf dry-matter content 3. LDMC may vary same leaves will be used for the determination of SLA. Garnier et al. LDMC may give more meaningful results. Section 3. tends to be inversely related to SLA and longer-lived leaves. These patterns are indeed often observed. you may be interested in the .g. Garnier et al. therefore. (2007). as described for SLA. Following the rehydration procedure. More on methods: see references in More on methods of Vendramini et al. First. Both within and involving SLA. More on methods: Wilson et al. Depending on the research question. because of relations and flammability (see Section 2. and its dry being thickest at the midrib. For example. the same leaves will be used for the determination of both Follow similar procedures as for Section 3. although conductance) via longer diffusion pathways. the equation simplifies to LDMC  1/(SLA  Lth). (2002). primary veins. see the case of xerophytic species in Section Similarly as for SLA. hail) N : area ratio). Lth and substantially during the day. wind. Ackerly et al. Most comments for SLA also apply to LDMC. higher in sunnier. 3.1 In many cases. (2001a). Each Thickness tends to vary over the surface of the leaf. Westoby (1994). and that full rehydration before measurement is compulsory. succulents have slow combination with lower internal transmittance. Vaieretti et al. (2008). hence. the leaf (rF). particularly if using a digital micrometer. many studies potential RGR and positively with leaf lifespan. Parkhurst and Loucks (1972). greater internal the two traits may not capture the same functions (this is self-shading of chloroplasts.1 and 3.7). Lth is strongly affected by LWC. References on theory. (2002). Niinemets (2001). This may in extreme cases be 10 times the value of a Special cases or extras single leaf. What and how to collect? What and how to collect? Follow exactly the same procedure as for Section 3. LDMC has been shown to correlate negatively with least in interspecific studies.1). balancing photosynthetic benefits against C Assuming that the fresh mass per volume of leaves is close to costs of respiration and transpiration. 3. Thick leaves growth. LDMC density  Lth) (where density = dry mass/volume  LDMC. Ryser et al.7). Measuring where any slight loss of turgor results in an underestimation. Garnier and Laurent (1987). as well as in LDMC. however.g. (1999. et al. Givnish (1985). Milla and Reich (2007). Royer et al. the strongest anatomical driver of variation in Lth tends to decompose more slowly than that from leaves with is the number and thickness of mesophyll layers. Poorter et al. (2007). or higher optical reflectivity in particularly obvious in some groups. leaf ‘work to formal relationship involving Lth and the average density of shear’ is (by definition) the product of Lth and tissue toughness. but not always.1). Leaves with high LDMC tend to be relatively Lth is a strong driver of leaf N per area. Commonly. Hodgson et al. Some aspects of leaf water studies. As is the case for SLA. (1999). Niklas et al. generally leaf sample is then dried in an oven (see Section 3. 2011). LDMC (and perhaps Section 3. Consequently. the SLA and LDMC. the leaves are cut from the stem and gently blotted dry with tissue paper to remove any Measuring surface water before measuring water-saturated fresh mass.1. In cases where SLA is difficult to measure (see species may have slower CO2 diffusion (lower mesophyll Section 3. mass subsequently determined.3). thicker leaves often have lower % LDMC. Poorter and Rozendaal (2008).1). Wilson et al. it is related to SLA by a strength of leaves (see Section 3. Using these units. are also a feature of succulents.190 Australian Journal of Botany N. drier and less fertile habitats. significance and large datasets: Eliáš Raunkiaer (1934). expressed in of SLA (see Sections 3. Cornelissen (1999). Pérez-Harguindeguy et al. Niinemets et al. module. Vile et al. for a combination of reasons.7). this relationship is often weak in interspecific than are leaves with low LDMC. (2002). as follows: LDMC = 1/(rF  SLA  Lth). significance and large datasets: References on theory. see also Section 3. divided by its water-saturated fresh mass (g). predicts that Lth should be 1 g cm–3. Lth also plays a key role in determining the physical of the leaf tissues. thicker-leaved environments. Kazakou et al. low SLA and low LDMC. because SLA  1/(tissue mg g–1. some form of rehydration should be seriously considered. at Lth. and are thus assumed to be more lead to faster photosynthetic rates per unit LA (via a higher resistant to physical hazards (e. see Appendix 1. species with low LDMC tend N and longer leaf-lifespan (which are associated with lower to be associated with productive.4 Leaf thickness Leaf dry-matter content (LDMC) is the oven-dry mass (mg) of Leaf thickness (Lth. e. Storing and processing Storing and processing Similarly as for SLA. Litter derived from leaves with high LDMC also among species.3). (2009).1). see is related to the average density (fresh mass per fresh volume) Section 3. (1999).1 In many cases. margins and leaf base. In laminar leaves. the have shown that outer-canopy ‘sun’ leaves tend to be thicker than strengths of these relationships are usually weaker than those those from more-shaded parts of the canopy. (2009). (2008). Second. (2005). (2001b). low LDMC. (2007). Witkowski and Lamont (1991). LWC is simply 1000-LDMC. mm or mm) is one of the key components a leaf. except that any dry storage should be avoided (however.12) also depend on covariance of SLA and %N. For recommended sample size. herbivory. Within individuals. Although higher Lth should tough (see Section 3. often highly disturbed photosynthetic rate per unit leaf mass).

or to compare replicates at a given point on the leaf. green leaves replacing the original contact points on the micrometer with should be used if in doubt and if decomposition is not the main contacts 2–3 mm in diameter. by dividing the total cross-sectional area by to herbivores. must not be mistaken for the actual pH of the leaf as a section. average Lth can be quickly estimated as whole or any part thereof.New handbook for measurement of plant traits Australian Journal of Botany 191 average thickness across the leaf. One method is to measure these quantities directly from such as C concentration or C : N ratio. unlike in living leaves. Poorter et al. see below under Special cases or A relatively fast approximation of whole-leaf average Lth can extras in the present Section) and is robust to differences in soil be obtained by dividing leaf fresh mass by LA (which is the same chemistry (including pH). varies substantially among species. On the positive side. LVDT). because substrate pH can be is relatively slow (e.e. this method enables leaf-tissue pH among species tend to persist during senescence. for soft- leaved species such as Arabidopsis.g. The cells of any plant tissue consist of three or more membrane-delimited compartments that can. or the are particularly strong determinants of leaf-tissue pH. Shifts in species composition.e. On the down making leaf-litter pH a worthwhile trait also. Wright and Westoby (2002). Differences in the section width. among Probably the fastest approach is to measure Lth using a dial. (2004). between major veins. whereas high concentrations of Other approaches are needed if one wants to distinguish organic acids and of C-rich secondary metabolites (chemical- between thickness of midrib. we recommend depends on the question to be addressed. however. If leaves are oven-dried before grinding. For example.g. although it can be useful as an ecological (i) Needle leaves. or both.g. at an intermediate position between the border and the midrib. and usually do. displacement transducer. soft tissue may distort when hand-sectioned. Still. It can be a reasonable side. Hodgson et al. (1999). several of different compartments. (2009). It will. near midpoint of leaf). litter pH values . indicative value of Lth for the feature in question (such as e. Parkhurst (1994). however. leaf. Still. narrow enough to fit issue. mixed together. which may actually ‘taste’ acidity. Enríquez reactions will have occurred long before the tissue is ground. (2005). e. Green leaf-tissue pH correlates positively (see Section 3. 15 min per measurement). significance and large datasets: compartmentalisation under conditions favouring pH-modifying Clements (1905). The variation is at least thickness over the entire leaf surface is to back-calculate it partly intrinsic (presumably genetic) because this pH can differ from the leaf volume divided by LA. i. have already stopped. Therefore.g. If necessary. Ground leaf-tissue pH integrates the as calculating 1/SLA  LDMC). by assuming that leaf fresh effects of many compounds and processes in the leaf that affect mass and volume are tightly related. and a decomposer community dominated by fungi. However. thickness measurements at several points in the lamina will measured by grinding up the tissue and extracting it with distilled be more appropriate. via the be made within quick succession and averaged to give an leaf litter. some substances into account the higher density of dry material in the leaf. Smith et al. divided by cross-section width). however. important to decomposers. as is commonly correlated (negatively) with important biogeochemical traits done. Garnier and locations or of special tissues. loss of References on theory. Multiple measurements can planting pines on high-pH soils dominated by bacteria can. at best. with a pycnometer. it is laborious greatly among different species growing in the same soil (and also to accurately measure leaf volume. Shipley (1995). However. during the day in CAM plants. potassium) will give high pH. (2011). Niinemets (2001). (e.1 for free software) to calculate average Lth with digestibility. water-extracted homogenate or ground material used for this method. but sufficiently broad so as not to dent the leaf surface when making measurements. or to use image analysis SLA (Section 3. be a weighted average Diameter  p/4 (equivalent to cross-sectional area of the pH values of the various compartments of the leaf. pH of green leaves. metabolic processes Díaz et al. and (positively) with leaf cross-sections (hand-sections). because needle possibly modified by reactions that occur between components leaves typically taper towards the leaf tip. approximation it works well.1). et al. when they are measurements would normally need to be made. the average of The pH of green leaf tissue and of senesced leaves (leaf litter). as an concentrations of metal cations (calcium. can drive changes in the pH of gauge or a digital micrometer (or even a linear variable the litter layer and of the soil’s organic horizon.5 pH of green leaves or leaf litter comparisons. This approach does not take its exchange capacity for H+ ions. or the thickness at special More on methods: Witkowski and Lamont (1991). lead to more acidic soil organic matter within decades. will tend to give a lower the leaf. at a position as standard as possible within the lamina Vile et al. have different internal pH values. reasonably accurate measurements to be made. the pH of the Special cases or extras type of crude. e. Whether midrib or lamina between the main veins) or region of interest green leaves or senesced leaves. When more precision is needed. permanent deformation is Physiological caveat difficult to avoid. Because. i.g. and between the tip and the base of the leaf. (1998). should be measured (e. Green and Kruger (2001). For needle leaves that are circular in cross- trait. pH. in leaf litter. species that differ in leaf-litter pH. avoiding important secondary veins) is acceptable for broad interspecific 3. the pH measured this way has been shown to also represent the Wilson et al. and the method proxy for litter decomposability. making it an important predictor of palatability across the section. Another way to estimate the average water. magnesium.g. High lower density as a result of intercellular spaces. (1996). Givnish (1979). Often one measurement per Laurent (1994). including vacuoles. The latter may explain why leaf-tissue pH tends to be half-way between the leaf base and the tip. margin and intercostal regions of defence compounds) such as tannins. Knapp and Carter (1998).

Grinding fresh leaves does not always work in ground plant material.1 Fresh green leaves can simply be ground or used as a tool to assess whether the availability of N or P is more chopped as noted below. Leaves used for sample. it them with steel balls in individual plastic vials on a roller mill. significance and large datasets: Zinke spectroscopy (ICP–OES). ground leaves tend to match those grinders are available. Effective. increasingly being replaced by methods that employ a (2006. See under Section 3.1 for discussion of when and whether Add distilled or de-mineralised water to the ground leaf to consider them. then following the above procedure. Petiole or rachis are often cut off before LNC and LPC analysis. Dry follow diurnal changes in leaf pH to indicate possible CAM the ground samples again for at least 12 h before analysis. (2010). we recommend chopping the leaf sample into Wet acidic digestion. you can carried out immediately before shaking and centrifuging. may be of interest to (additionally) measure fresh leaves. as long as method and equipment to be applied (~2 g dry matter per replicate calibration is adequate (using buffer solutions of pH 4 and pH 7). e. Kjeldahl digestion for N analysis is (1962). Leaves collected for leaf SLA limiting for plant growth.6) and stored See Section 3.e. followed by colorimetric (flow-injection) analysis ball mill. growing at the same site. The supernatant can then be measured for pH by leaf material per replicate is collected. LA or SLA analysis can be also used to measure LNC and LPC. Grind each replicate Special cases or extras separately. store the material air-dry and in the dark until use. because pH values obtained from oven-dried leaves are largely comparable to those from fresh leaves.1 for the leaf-collecting procedures. we recommend adding 1. any petiole or rachis should be removed species tend to vary significantly with the N and P availability in before pH analysis. Marschner (2012). for various specific purposes. matter into N2 and CO2. Macro.12). to a maximum of 1 year. per unit of dry leaf For green leaves. then provided drying temperature has not been higher than 70C. Oven dry at 6070C for 72 h. measure P by inductively coupled plasma–optical emission References on theory.6 Leaf nitrogen (N) concentration and leaf phosphorous (P) sampling. Alternatively.5-mL Eppendorf Section). see Section 3. expressed in mg g–1 (or sometimes as % dry-leaf mass). however. see below under the present water to 0. Avoid cell contents of which have remained intact until shortly inter-sample contamination by cleaning the grinder or steel balls before measurement. i. but are included in some other Measuring cases. LNC and LPC of a given As a default option. the which is an efficient way to grind many samples at once.2 mL of case of combustion techniques. has been widely used. Germany) fits nicely inside such Eppendorf tubes. for N and 5 g for P in the case of acid digestion. 2011).1). Instead. after being oven-dried to obtain legumes. Samples may also be ground by shaking on fresh leaves fairly well. (2006. make sure Interspecific rankings of LNC and LPC are often correlated.192 Australian Journal of Botany N. inexpensive mechanical measurements on dried. This should be precise method for measuring total P. Use a ball mill for small samples. Initial air-dry until the analysis. Pérez-Harguindeguy et al.2 g for N in the If samples are small in volume. should reflect the actual situation in the litter at the time of 3.15 mL of ground leaf material in a 2. Storing and processing Weilheim. enough of them are collected so as to get enough material for Across species. concentration Leaf N concentration (LNC) and leaf P concentration (LPC) are What and how to collect the total amounts of N and P. Air-dried leaves or leaf What and how to collect? litter are ground as for leaf N analysis (see Section 3. Cornelissen et al. converting organic More on methods: Cornelissen et al. see Special cases or extras in their environments. many analysis can instead be used. rehydration is not necessary. Finzi et al.g. see under Section 3. Related to this. (using different reagents for N and P). Freschet et al. A thin SenTix 41 electrode connected to an Inolab level 2 pH meter (both WTW. However. LNC tends to be closely correlated with mass- the analysis. Initial leaf rehydration is not necessary. Actively N-fixing species. tend to have higher LNC : LPC ratios than other plants their biomass (see Section 3. but make sure that enough total supernatant. 0. procedure before processing. followed by mass spectrometry or gas . making cleaning between samples laborious.or micro-Kjeldahl (acidic) well because solids may stick to the surfaces or to the balls in a digestion. respectively. followed by formation of blue ~1-mm-diameter pieces with a razor blade or an automatic phosphomolybdenum complex from orthophosphate is a more chopper that gives comparable fragmentation. Shake the samples in a laboratory rotary shaker for 1 h. 2011). (see Section 3. The LNC : LPC (N : P) ratio is sometimes Section 3. High LNC or LPC are generally associated with high nutritional quality to the Processing and storing consumers in food webs. Although ones (repetitive strain injury). but is not recommended for large (1) Additional measurements on fresh leaves. Manual grinding with mortar and pestle is an option for small numbers of samples. If leaves are small. combination of combustion analysis. (1998). For instance. according to the analytical using any of a wide range of laboratory pH meters.1. this may be useful to carefully between samples. leaves of CAM plants are Measuring best collected in the afternoon. tube. After oven-drying the leaves. in comparisons of CAM plants with other plants for leaf pH. For centrifuge until there is a clear separation of the sediment and the replication.1 for the collecting and storing mass. and subsequently stored air-dry. to give an 8 : 1 volume ratio of water to leaf sample. based maximum photosynthetic rate and with SLA. well after nocturnal acid Several techniques are available to measure LNC and LPC accumulation.

g. recommended sample size. References on theory. Ws. We recommend running Section 3. Measure according to the direction in which the force is applied. Very small leaves. herbivory. very width. divided by its to the edges of the clamps could help.1. expressed in N. see Appendix 1. For unit conversion. with automated N analysers. or its the leaf tears. See Section 3. expressed in N mm–1. hail) and biotic (e. Storing and processing For leaves too tough to be torn. NO2–. remember that leaf or leaf fragment (expressed in N mm–1). fragment to obtain Ft. by putting them between sheets of absorbing paper in a plant press. Aerts and Chapin (1999). if samples measurements on epidermis fragments. if the focus is on chewing (1) Leafless and heterophyllous plants. because some N fractions (e. There are some more sophisticated instruments to measure Ft. for Mecmesin Ultra Test Tensiometer (Mecmesin. Temminghoff and Houba (2004). For analyses. which would be for species with soft leaves. MA. see Section 3. (2004). Ft is likely to be the most meaningful property. carry out the strength for several days. It also tends tender species). A thin piece of rubber added the force needed to tear a leaf or leaf fragment. sometimes called ‘force clamped tightly without much tissue damage. Slinfold.1. see give reasonably accurate LNC and LPC. unless the latter is not obvious (e. Grime et al.4). we recommend measuring the property that is most closely related to the process of interest. with increasing force. and if carried out leaves. such as What and how to collect? the 5542 (Instron. contributing to longer leaf lifespans. reflects the mean force needed to cut a leaf or leaf double-sided tape. USA) or (with adaptations) For the selection and collecting procedure.1 and store (down to 1 mm if necessary and possible). have to be sent to distant locations). Leaves too tender to provide an actual measurement with the apparatus are assigned an arbitrary tensile strength of zero. leaves in a cool box or fridge.g. rehydration At least five instruments have been used to measure work to is needed before measuring. They all measure how much work is required to cut a . (1997.g. If still too tough. To proceed. significance and large datasets: Chapin (1980). proceed to measurement straight away. cut a leaf fragment from the central section of the leaf but away from the midrib (central 3. (1) Tearing (tensile) tests Aerts (1996). However. first rehydrate by wrapping in moist paper and some cyclic N compounds) do not react in Kjeldahl analysis. are generally less labour. Try to do this force to tear (Ft). Combustion techniques also generally recover more N than do Kjeldahl For fresh samples. If you have the choice. we recommend using to have afterlife effects in the form of poor litter quality for fragments with a length : width ratio between 5 and 10. 2010). some Poaceae or Liliaceae).g. different ways. work to shear tests would be the best approach (Table 2). we believe that all of these standard methods should may be better for some xerophytic. we describe three methods that have produced a standard reference material with known LNC and LPC along good results and for which purpose-built equipment is available. with a simple apparatus that includes a 0–3-kg-range Horneck and Miller (1998). Field and Mooney (1986). Toughness of fresh and rehydrated shear (Ws). work to shear (Ws) and force to punch (Fp). Both Ws and Fp are 1 kg = 9. If this is not possible (e. or the leaf is too small for doing so without using a Physically stronger leaves are better protected against abiotic magnifying lens. UK).g. the the exact width of the leaf sample before placing it in the physical strength of the leaves can be defined and measured in apparatus. Express the result in N mm–1. NO3– and air-dried samples. Watch the dynamometer to read the force at the analogue. an alternative is to air-dry the (2) Shearing (cutting) tests samples. Canton. Ft is gently without damaging the tissues. in the cases of both Ft and Ws.and Measuring chemical-intensive than are Kjeldahl analyses. Note that Ft has been previously tough and slippery leaves. Lambers and Poorter (1992). to decomposition. immediately after collecting.81 N. dynamometer (Fig. put in a sealed plastic bag in the fridge for 24 h (gentle spraying However. (1997). Anderson and Ingram (1993). Force to tear (Ft) can be easily and inexpensively measured More on methods: Allen (1989). These combustion techniques provide leaves is well correlated for sclerophyllous leaves and grass concentrations of both N and C in the leaf. Because leaves have different strength properties make sure that force is applied along its main axis. Then pull slowly. The three most common measured properties are Then fix both ends of the sample with the clamps. (1997).1). certainly within 72 h highly succulent leaves (or modified stems). first try a narrower sample Follow the procedure described within Section 3. by using strong of fracture’. Whenever possible.7 Physical strength of leaves vein). with the samples. Divide the total force by the width of the leaf strongly influenced by Lth (see Section 3. until fragment at a constant angle and speed. apparatus (assuming sample width of 1 mm). insects or trampling by mammals.New handbook for measurement of plant traits Australian Journal of Botany 193 chromatography. and slightly succulent leaves may be referred to as ‘leaf tensile strength’. In this case. Reich et al. then tensile strength equals the maximum possible value in although it may be desirable for accurate measurement of Lth. Tougher leaves tend to keep their squashed if clamped into the apparatus. wind. In the case of Special cases or extras herbivory by vertebrate grazers. The length of the fragment follows the (e. Cornelissen et al. Rehydration is not indispensable. 3a). J m–1. In the case of Measure as soon as possible after collecting. Wright et al. Physical leaf or leaf fragment depends on the tensile strength and tends to investment in leaf strength is a good indicator of C investment vary between 1 mm (extremely tough species) and 10 mm (very in structural protection of the photosynthetic tissues. The width of the damage. Here. Fp is the force needed to force a punch through a moment of tearing. Place it perpendicular to the edges of the clamps. rotting-sensitive species. trampling) mechanical longitudinal axis (direction of main veins).

from Aranwela et al. snails). identification of plant resource-use identification of plant resource-use decomposition. Bucks. Pérez-Harguindeguy et al. USA (Onoda 4202. 1991) High Wycombe. Ws Leaf resistance to a punching force. Canton. Shimadzu DCS-5000 Bucks. MA. (b) scissoring test (prototype model from Darvell et al. (Lucas et al.194 Australian Journal of Botany N. 3. universal et al. 1996. trampling). as described in Hendry and Grime (1993). Instron. Instron. decomposition. a very similar device is described in Wright and Illius 1995). (c) shearing tests reproduced from Wright and Cannon (2001) (safety guard. decomposition. Different apparati for measuring physical strength of leaves.g. herbivore impacts insects. Types of tests commonly used for measuring leaf mechanical properties Parameter Tearing test Shearing test Punch test Property measured Leaf resistance to a tearing force. on vegetation (e. and (d) punch tests. UK (Wright and Illius 1995) Study objective Herbivory by mammalian grazers and Herbivory by chewing insects and Herbivory by chewing or sucking other tearing herbivores (e. Fp Unit N mm–1 J m–1 N mm–1 Apparatus Tearing apparatus (Hendry and Grime Leaf-cutter machine (Wright and Universal testing machine: 5542. High Wycombe. UK (Wright and Illius 1995) testing machine: 4202. identification of strategies strategies plant resource-use strategies (a) (b) (c) (d ) Fig. small vertebrates. scissoring machine Instron.g. Diagrammatic representation of the devices used for (a) tearing tests. 2008). 1996). Table 2. (Darvell et al. . quick-release sample holder and electronic control unit not shown). Ft Leaf resistance to a shearing force. universal testing machine: Cannon 2001). (1999). 1993).

Then.New handbook for measurement of plant traits Australian Journal of Botany 195 leaf. Measurement of Lth (in mm) device for measuring the average force needed to fracture a for this purpose is most practically carried out with a calliper.neoninc.5 mm) attached to a spring-loaded balance or a be spread out over a much longer period. This measure constitutes one Consistency across the leaf tends to be reasonable. either with a single blade against an anvil. This method has been more widely used in the display outside the main foliage peak of the more competitive tropics. A non-portable apparatus for this measurement is showed before placing it in the measuring apparatus. Lucas procedure according to the apparatus you are using. as long as important component of the timing of plant growth. Species with longer-lived leaves tend to invest Garnier. in which case a plant counterweight (being a container gradually filled with water and may maintain several cohorts of leaves of different ages weighed after penetration). must frequently clean the blade(s) with an alcohol-soaked wipe. Ws or Fp by the (average) blades (‘instrumented scissors’). Kitajima that this methodology is highly sensitive. considered a mechanism to conserve nutrients and/or reduce A calibrated copy of the apparatus described in Fig. leaf at a constant shearing angle (20) and speed is detailed in immediately after measuring the leaf-sample width and in contrast.cnrs-mop. be sure to complete the calibration Wright and Vincent (1996). the apparatus consists of a mechanical References on theory.05–0. Certain groups of competition avoiders (including sufficient. A clearance of 0. (1) Leafless plants. accessed 22 February 2013).usanpn. before the canopy of the materials (such as 5542.php. at the widest point along the lamina Leaf lifespan (longevity) is defined as the time period during (or halfway between the base and the tip. one can remove the or years. Leaves are cut 3. This is because leaves may senesce all all of which have some kind of fine-needle or a flat-ended punch together (as in deciduous species). . if this is difficult to which an individual leaf (or leaf analogue) or part of a leaf determine). Three measurements per leaf are probably phenology. A sharp edge in the flat end of the punch will also avoid Thus. vibrations. (particularly the epidermis) to rupture. (2000). or big veins are avoided. defined by the number fraction length. die. Wright and Cannon (2001). Aranwela et al. This test does not work well for many grasses and some gap colonisers) may have very short periods of foliar other monocots. A portable and widely used thickness of the leaf sample. (1999).org/ interesting additional parameter of leaf strength can be budburst/index. and a plant. 3c. To standardise the force per unit to measure the duration of green leaves. the midrib may be removed so that (see Monocotyledons below in the present Section) is alive only lamina tissue is tested (or lamina and midrib are tested and physiologically active. (2011). as is the case in many temperate evergreen hole in its centre. the operator must avoid and Poorter (2010). email: garnier@cefe. some tropical evergreen species maintain and the edge of the hole in the die is recommended to avoid error in a continuous green leaf canopy with a very short leaf lifespan. photosynthetic unit) is green. Different penetrometers of growth or the proportion of the year when a plant is able to (Fig. significant resources in leaf protection and (partly as a consequence) grow more slowly than do species with short- (3) Punch tests lived leaves. Long leaf lifespan is often refer to the specific user’s manuals). or their senescence may (diameter ~0. Choong et al. Cornelissen et al. 3c is respiratory costs in habitats with environmental stress or low available for use at CNRS in Montpellier. (2004). excluding toughness Leaf lifespan does not necessarily indicate the phenology provided by midribs and main veins. Onoda et al.1. To proceed. Instron). to understand the ability of a plant to exploit its seasonal overestimating toughness values resulting from compression and light environment and the timing of its growth. file is recorded. thus providing an indirect index of important plant analyse these data separately. months separately). France (contact Eric resource supply. Darvell et al. before placing it in the apparatus. a power source and a computer in which the output (1988). Detailed Special cases or extras information on monitoring all the aspects of plant phenology can be obtained from several networks set up for this purpose. it is also useful tension rather than shearing. seasonally green species closes. (1992). The punch goes through a die with a simultaneously. In both cases.5–5. This February 2013) and Project Budburst (http://www. accessed 22 (2) Leaf-tissue toughness or leaf-specific toughness. they also conserve internal nutrients longer. The litter of (previously) long-lived leaves tends to be relatively Force to punch (Fp) is the resistance of the actual leaf tissues resistant to decomposition. (1996). Measure the exact thickness of the leaf sample (lamina or midrib) with a calliper. 3b.g. Alternatively (and less precisely). some spring geophytes manage important growth at the more sophisticated apparati designed for measuring properties of beginning of the favourable season. Díaz et al. In addition. many evergreen species have a year-round ability to photosynthesise. nutrient-use efficiency values can also differ between apparati (for calculation process. It is expressed in days. and decomposability of litter.1 mm between the punch species. Turner (1994). measurements as a result of friction between the punch and the because they have a high and continuous rate of leaf production. significance and large datasets: Coley portion.8 Leaf lifespan and duration of green foliage at right angles to the midrib. Leaf lifespan relates to the nutrient-use strategy of portion of the data that represents the midrib being cut. including the NPN (http://www. In some studies. or with a pair of obtained by dividing Ft. Considering et al. Grubb (1992). 3d) have been used in the past (there is no standard design). wind). one has to divide the force by the circumference of months per year that the leaf canopy (or analogous main of the punch. and More on methods: Hendry and Grime (1993). perform photosynthesis. any possible source of external noise (e. in Fig. place the sample in the anvil and fix it with the clamp. Read and Stokes (2006). The procedure to calculate final traits such as potential growth rate. (1999). See Section Some recent studies have added the punch and die to species. The data are therefore expressed in N mm–1.

The leaf-identification individuals and leaves are the same as in Method 1 information needs to be recorded on the leaf itself (or above. the estimate of the median lifespan of different cohorts. rather than the Picea. it is (such as colour and/or symbols). or interannual variation is likely to cause shifts in the however. although most died. Periodically revisit and first time at a census event. although in an unambiguously (4) Counting ‘cohorts’ for many conifers and only some systematic way). some species time interval. can be made in which different method is very easy and quick. and most mortality occurs in the year (see Appendix 1). Method 1 (below) is the best. Tag or map individual lifespan. For instance. This can be tricky. This and leaf drawing. Care must be taken to make these interval length increases relative to the mean lifespan. We recommend about eight counts over this that in Mediterranean-type climates. by using the same criteria as in Section 3. it is simple to first census when a leaf is no longer present (or is visibly count. for plants in their exponential growth phase. In all cases periodicity occurs) and the branch or shoot is in quasi- where a general measure is desired. In addition to providing the most younger cohorts. measures for multiple consecutive cohorts. or map. or (2) if one estimates %mortality interval. At the marks on the shoot or branch. and for Methods 2–4 can replace Method 1. but can be used only colours are used at each census event. Number of individuals and leaves are the (1) Periodic census of tagged or mapped leaves. Pérez-Harguindeguy et al. woody pioneers in (A) Dicotyledons. This method intervals of roughly 1/10 of the ‘guesstimated’ lifespan) is effective when many leaves are produced within a whether they are alive or dead. For one gets to the cohort with less than 50% of the original each individual plant. This is a good method if the census is and shed them all within a short time period (see . and usually no or very few survivors accurate estimate of the mean and median leaf lifespan. upper canopy) from at least 10 individual plants. Because the census short time period. This this method also provides a frequency distribution and method can also work (1) if there is some mortality in enables estimates of the variance. Calculate may flush more than once per year. although it is temperate zone or many herbs. branch by branch. we recommend of peak ‘turnover’. Note that a total of leaf lifespan. This period can be much shorter for fast-growing plants such as tropical rain forest pioneers. the birth and death of individual leaves over time at (3) Observe a cohort of leaves until one-half have repeated census intervals is the best. which most other younger cohorts and a roughly equal proportion of methods do not provide. and often is done with a brief code woody angiosperms.1. although some Pinus species number of branches or shoots per individual. but a higher frequency may be better in experience two growing seasons each year. Leaves each successive cohort can be identified either by produced in given census intervals are drawn in such a differences in foliage properties or by scars or other way that their relative spatial positions are clear. select individuals and leaves equilibrium in terms of leaf production and mortality. a branch important to be very familiar with the species. In that case. if seasonal the accuracy of any individual measure will decrease. more quickly). number of leaves produced and died over a time interval some conifers may appear to be missing needles that represents a period of apparent equilibrium for leaf (judging from scars) that were never there in the first production and mortality (see below within the present place because of reproductive structures. Many conifers. with a large sample size. Count (for each shoot or branch) the total cohort by cohort. production number and the accumulated leaf-mortality (5) Duration of green foliage for species that produce most number (facilitated by plotting leaf production and leaf of leaves in a single ‘cohort’ within a small time period death against time). This technique is useful most labour-intensive and takes a longer time period. survivors in cohorts older than the first cohort with (2) Count leaves produced and died over a time >50% mortality. nearby on the branch. Be aware Section). This works if there is little leaf mortality for at least 40 and ideally 160 leaves per species is necessary younger cohorts. Then estimate mean leaf lifespan as the shoot (preferably the leader or a dominant shoot in the mean distance in time between the accumulated leaf. Number of mean should remain accurate. because there is some mortality in per individual plant. Tracking same as in Method 1 above. For woody angiosperms. Count one some cases. In this method. This method measures the median leaf labour-intensive method. it is crossed out on the more than 50% of the original foliage remaining until drawing with the colour of the present census event. To achieve this. but only if the plants with very long leaf lifespan (because one gets data criteria are met. and record periodically (at count the number of leaves remaining. show this pattern. Measuring leaf lifespan long enough to cover seasonal periodicity (so typically it Different methods are required for taxa with different kinds of needs to be several months up to a year if seasonal phenological patterns and leaf demographic patterns. and use that as the estimate of mean shoots and sample all leaves on them. select two or more branches or foliage remaining. Alternatively.196 Australian Journal of Botany N. This method gives a the lifespan for each individual leaf and take the average slight overestimate. especially Pinus and increasing the number of individuals. the if the species is known to produce foliage at regular position along the branch that separates leaves produced known intervals (most frequently once per year) and in consecutive census intervals must be marked. the number of cohorts with mostly or completely dead). in the cohorts older than this ‘peak turnover’ one. Count the number of leaves that appear leaves (not leafy cotyledons) when they appear for the between two census events.

Westoby et al.9 Vein density soaking time in all solutions. 1997. ensuring a large enough field of view (e. showing both broad phylogenetic trends and References on theory and significance: Uhl and Mosbrugger potential adaptation to resource gradients. see Section 3. making the mean of the leaf in H2O and transfer to a 2% w/v NaOCl–H2O solution for the whole blade lifespan much longer than the lifespan of a 5–30 min. and divide this number by the image area to obtain the vein individual leaf lifespans can be as short as a few months. (1) Leafless plants. In some species. if possible. not particularly Rinse the leaf in H2O and transfer via dehydration series to meaningful as a measure of tissue longevity. 2010). Leaves become delicate during processing References on theory. this may not provide an estimate of green-tissue longevity that is comparable Soak the leaf in 5% w/v NaOH–H2O for 24–72 h. (2004). Then rinse old tissue becomes senescent over time. Take particular care species in the survey at least once a month during the favourable to ensure that all veins in the network are visible. 30%. 70%. below in the present Section). until it becomes bleached.g. If photosynthetic tissues do not die and fall off as separate units. in immersion oil Observe the foliage of 5–10 individuals of a given species or Permount (allow several days after mounting for toluene to several times throughout the year. Brodribb et al. (1999). use the entire leaf. shows plasticity across environments and is highly variable among species. This method is likely to be the least lamina of given leaves. or Measuring duration of green foliage permanently. so that leaves will not Aerts (1995).1). Stain the leaf for 15 min in 1% w/v safranin O in adaptation of Method 2 above. Sack and Frole (2006). significance and large datasets: Chabot and should be moved carefully between solutions or solution and Hicks (1982). (2000). until it to the measures above for dicots. very thick or dense leaves may require several days in the NaOH solution before Vein networks constrain the transport of water. C and nutrients they become transparent. Reich et al. as specified above for monocotyledons. Vein counts should be made using estimated to have at least 20% of their potential peak-season microscope objectives of 4 for ferns and up to 40 for foliage area are interpreted as ‘green’ months. leaves may have sclereids that Vein density is a structural determinant of hydraulic conductance can be mistaken for veins. hydraulic function. after transfer to 100% toluene. one during the middle hard to see. 2004). puncture or tear. particular section of the blade and. Wright et al. Warm More on methods: Jow et al. protocol. below in the known trait variation between sun and shade leaves and across the present Section). or more concentrated NaOCl–H2O solution for a shorter time period. replace the soaking solution. and 5 min for can be assessed to estimate the tissue longevity. or 50%. The months in which the plants are accurate visualisation. Most species with a light microscope. The most season (preferably including a census shortly before and shortly numerous and functionally important minor veins can be very after the favourable season) and. 100% ethanol. (2) Sclereids. 2004). each step lasting 30 s. for larger leaves. the details of the protocol can be modified. Coley (1988). If possible. therefore. Depending on the species set considered. For smaller leaves. for tender leaves. measurements should be made reliable of the five described herein. follow Method 2 (above) for specific Special cases or extras zones of the photosynthetic tissues. . We recommend a census for all fully evaporate). Roth-Nebelsick et al. 100% the production and mortality of specific zones of the blade ethanol. Small or thin leaves may require less 3. If the NaOH solution turns an opaque brown related taxa. ethanol and/or other lignin stains. Then measure the total length of veins in the image Note that in some evergreen species from the aseasonal tropics. (2002. the blade continues to grow new tissue while colour during soaking. Destain in ethanol. (1980). with an tougher leaves). the length of minor veins per unit of epidermis. Craine NaOH may be used (although not boiling). density. and often require the epidermis to be removed for of the unfavourable season. However. Wright et al. 50%.New handbook for measurement of plant traits Australian Journal of Botany 197 Measuring duration of green foliage. given the growth habits of many monocots Measuring (see below within the present Protocol). (2001). longevity of entire blades can be measured as described above. This trait and function (see references below in the present Section). For best results in clearing leaves of given Poorter and Bongers (2006). Conversely. pure ethanol (e. the sufficient. Veins will appear red. Diemer (1998). Boyce et al. For some monocots species. This image analysis process may be semi-automated with freely Special cases or extras available software (see More on methods. This census can be tropical angiosperms. species. For the leaf collecting and Holbrook (2006). on sections of lamina containing no large veins. (2007. Sack Fresh or dried leaves may be used.g. In some grasses and becomes transparent. Reich et al. (1) Leaf handling. using image analysis software (see Section 3. Hairy leaves may require removal within the leaf. (1992. Additional venation traits can be the vein density may correlate with other leaf traits. In such cases. (2002). and longer et al. LA (mm mm–2) can characterise the structure of these networks. such as Lth. The leaf can be mounted in water or glycerol on plastic transparency film. should be vacuumed out of the dish. individual leaf lifespans >1 year will be green throughout the year. and also have an important and photosynthetic rate. measured for more detailed investigations of leaf structure stomatal density and maximum gas-exchange rates. a 1-cm2 section is (B) Monocotyledons. Vein density. 1–10 mm2). but the accuracy should be tested. Photograph the venation network under combined with assessment of Leaf lifespan.1 Sampling intensity should account for (2009). clearing times. (1999).

Amax scales with other structural. along with those other variables. aspects of the leaf economic spectrum and. range (next paragraph) can help ameliorate this problem by Measurements can also be made on detached foliage. photosynthetic temperature optima. source–sink inhibition or other causes. Because most published measurements have photosynthetic rate (see Section 3. enables comparison among species and especially among sites and times of year is challenging. direct sun for 5–10 min) to minimise canopy processes of whole ecosystems. and tests of intact v.1).g. that is also useful. If possible. In any case.198 Australian Journal of Botany N. However Rleaf under What. More on methods: Dilcher (1974). to a few minutes at most. focusing on the shaded taxa of understorey. when and how to measure? typical field conditions is valuable because it is both a measure Sample young.10). concerns about leaf induction status or stomatal closure as a result of shading (see Section 3. unless that is the focus of of gas-exchange responses of the taxa under study will be the research.1). limited Sample young to medium-aged fully expanded leaves soil moisture or air humidity. and in the dark until measurement. Some knowledge e. 3. Price et al. irradiance and to minimise variation resulting from very recently fixed temperatures are near optimal for the taxa in question. 1985b. Do not make in choosing the time of year. Detached leaves should be kept moist. See under Section 3.10 Light-saturated photosynthetic rate If possible. water stress). Simultaneous measures of leaf water. Gardner (1975). therefore. Pérez-Harguindeguy et al. Sample foliage from parts of the canopy sunlit essential. detached foliage should be made for a subsample. If possible.g. Do make measurements on days when soil night. after detachment. Conduct some (which may hold more foliage) and/or choosing a standardised test comparisons of gas exchange on intact and ‘re-cut’ branches. the leaf material inside the chamber should be The light-saturated photosynthetic rate (Amax) under typical collected (see Section 3. Wright et al. or been made under ambient CO2 concentrations. More on methods: Wong et al. are cold relative to for any subsequent leaf handling. (2004). (2011). Because realised photosynthesis is less than maximal because What. check reliable measurements. significance and large datasets: Reich least as an index) of metabolic capacity and a factor determining et al. enables scaling to canopy processes of whole ecosystems. however. heat or cold stress. chemical and longevity aspects of the leaf economic only if they have been in sufficiently high light just before spectrum and. 1985c). unless one is specifically focussed on the (days to weeks) periods of severe water deficit. those for Amax and. 1985a. air humidity. closure. usually expressed in mmol m–2 s–1 or stored for any subsequent chemical analyses. Do not make measurements during or just following during daytime. and general conditions measurements during or soon after atypical conditions (such as under which measurements can be made. 1991b). Ellsworth and Reich (1992). using specialised chambers whether given individuals fail to stay hydrated. 1997. Measure leaves structural. or mornings in general. within seconds. enables scaling to measurement (e.10 morning. detached foliage should be made for a subsample. or unusual shaded understorey taxa. chemical and longevity Reich et al. measured for LDMC and SLA. given the sensitivity and acclimation of Rleaf to temperature. measure intact foliage or else leaves on this problem. to ensure similar rates are observed.1) to ensure negligible respiration associated and source–sink inhibition. and portable photosynthesis system may be suboptimal for taxa conditions in the chamber can be either set at levels considered with lower flux rates. and field conditions. (1991a. recommended. however. temperature that is at the high rather than low end of the candidate to ensure the technique works for your taxa and system. nmol g–1 s–1. Brodribb this requires even greater attention. foliage. to alleviate optimal). merely by turning off.1 for discussion). 1999). that would be shielding. If a given to ensure that similar rates are observed. 3. One can reduce flow rates and/or optimal or left to track the in situ conditions (which need to be near increase the amount of foliage into the chamber. Rleaf scales with other metabolic. among others. (1979. the chamber completely from incident light. If possible. However. . Measurements in most ecosystems should be made at mid. care must be taken with biosynthesis (‘growth respiration’). average realised photosynthetic rate (for upper-canopy foliage). In the latter case. measurement can be made Leaf dark respiration (Rleaf) can be measured while measuring later in the day. along with those other variables. In such cases. this is not always sufficient to obtain branches cut and then re-cut underwater. leaves must have been in the dark for ~30 min Any reliable leaf gas-exchange system that can control leaf late morning (e. from 0800 hours to 1100 hours local time) temperature can be used. unless specifically night-time respiratory C flux. or both. fully expanded leaves (see Section 3. (1992. when and how to measure? of a host of factors. including low or high temperatures.11 Leaf dark respiration vapour conductance are typically made in concert with the Characterising leaf dark respiration (Rleaf) in a fashion that photosynthetic measurements. negative leaf water potential. measure intact leaves at temperatures. the signal to noise ratio of the typical Any reliable leaf gas-exchange system can be used. These of basal metabolism and a rough correlate of average realised should be from sunlit parts of the canopy. If rates can also be measured under saturating flux rates for Rleaf are roughly an order of magnitude lower than CO2 concentrations. Leaves should be measured and Feild (2010). is a valuable metric as a measure (or at References on theory. time of day. increasing the flux rate. plant water status. cool (to minimise This minimises the risk of sampling during midday and C and water loss). it is best to measure intact under non-limiting vapour-pressure deficits or temperatures. (see Section 3. afternoon declines in gas-exchange rate as a result of stomatal tests of intact v. If possible.g. photosynthate or transient light-induced respiratory CO2 losses.

significance and large datasets: Reich portion of a herbarium specimen. using a micro-wave and then be dried as quickly as possible at measure (subsets of) leaves at appropriate contrasting 70–80C. to avoid changes caused by loss of organic matter measurement temperatures. store at least part of the samples fresh (see spread over the entire mesophyll (photosynthetic tissues).1). 10C or 15C in cold boreal or summer tundra conditions). d 13C alone (for facultative CAM plants. C4 Carbon isotope ratios of organic material (d 13Cleaf) are plants tend to perform well in warm. best seen in a cross-section. d 13C values have been intraspecific variability is small enough not to interfere with found to range as widely as from –14‰ to –23‰). submerged aquatic plants have CAM too. water. However. 20C in the temperate zone.3 ‰. and the many plant families. C3 photosynthesis d 13 C : 21‰ to 35‰.g. instantaneous its isotope composition.g. Although least four different temperatures over a 1535C range. and Measuring responsiveness to elevated CO2 Compared with C3 plants. C3 plants have leaves in which anatomical analysis (see under (B) Anatomical analysis in the all chloroplasts are essentially similar in appearance and present Section). However. CAM can be inexpensively confirmed by verifying that C4 photosynthesis : 10‰ to 14‰. 25C in the tropics. (2001). There are obligate parts per thousand.03 ‰ and 0. Separating C3 or C4 from CAM plants is difficult on the basis of intraspecific genetic differences and/or phenological conditions.e. Wright et al. Where possible. If this is not possible. Using a razor blade or Collect the fully expanded leaves or analogous photosynthetic microtome. then the plant found. (1998. Bulk the replicate leaves or tissues for each plant. including C3. Once dry. or salty environments (e. (2010).12 Photosynthetic pathway screen. stomata are open at night and closed during the day. including the midrib and three leaves from each of three individual plants. or CAM v. or by measuring diurnal patterns of organic acids or leaf pH values. or ideally at (through leaf respiration or microbial decomposition). in tropical savannah-like ecosystems). Although C- isotope composition can be affected by environmental factors. Some Belemnite (PDB) standard as d13C in units of per mil (‰). Fig. only C3 metabolism has been found. We recommend sampling at least (particularly thick and protruding veins. Two main identification methods are available. (2005). Only small amounts of tissue are each with their particular biochemistry. then grind the dried tissues thoroughly to pass through a 40-mm-mesh or finer 3. C3. sunny and relatively dry and/ measured using an isotope ratio mass spectrometer (IRMS. The Section 3. mesophyll cells are not concentrated around the veins and are . These pathways have important consequences for 3 mg of dried organic material is used. dependent on the IRMS whereas CAM plants are generally very conservative with water used) and are traditionally expressed relative to the Pee Dee and occur predominantly in dry and warm ecosystems. Be aware that insecticides or et al. because Dry the samples immediately after collecting. It is photosynthetic tissue is succulent or organic acid concentrations useful to know in which families C4 and CAM have been are high during the night. In rule of thumb. depending on environmental factors following (see graphic explanation in Material S4. After isotopic analysis. less than CAM. make cross-sections of leaf blades of at least three structures of adult. In most cases.1) or killed by respiration across temperature regimes. 2008). so that species from those families can be screened is CAM. the cross-study comparisons are often made of taxa grown in sample can be stored for long periods of time without affecting and/or measured under different temperatures. as well as on the aim of Obligate CAM : 10‰ to 15‰: the work (e. as a the distinction between C4 and C3 photosynthetic pathways. If conducting major laterals.and nutrient-use efficiencies. are not relevant). optimum temperatures of photosynthesis (higher in C4 than in C3 plants). samples can also be collected from a References on theory. healthy plants growing in full sunlight or as plants per species. (B) Anatomical analysis C3 and C4 plants typically show consistent differences in leaf What and how to collect? anatomy. In such cases. It is often easier with small samples to grind all of the Three main photosynthetic pathways operate in terrestrial plants.New handbook for measurement of plant traits Australian Journal of Botany 199 For comparative Rleaf measurements.g. Below we describe two methods which in be decisive (see (B) Anatomical analysis in the present Section). if d 13C is between –10‰ and –23‰. combination provide good contrast between pathway types. with 10C intervals. C3).g. other sprays that may have been used to preserve the specimen. anatomical observations and diurnal systematically as potential candidates for these pathways (see measurements of gas exchange or biochemical analysis would Material S4. (2004). Table 1). not the preferred procedure. epiphytic orchids in high-elevation Australian rainforests). C4 and required for a C-isotope-ratio analysis. Tjoelker et al. Facultative CAM : 15‰ to 20‰ and Which method to choose (a combination would be the most reliable) depends on facilities or funding. Atkin et al. 1): (e. material with mortar and pestle. i. can affect its isotope composition. precision between 0. namely C-isotope composition and anatomical observations. Rodriguez-Calcerrada et al. the photosynthetic CAM species and also facultative ones. which may switch pathway of the species can be determined on the basis of the between C3 and CAM. but low during the day. one typically chooses (A) C-isotope analysis a standardised temperature appropriate for the site conditions Storing and processing (e. to contrast C4 v. the sample should temperature response functions can help in calibrating first be stored moist and cool (see under Section 3. making sure to include some regular veins close to full sunlight as possible.

As a result. The cells directly surrounding the (1990). (1989). D = a + (b – a)ci/ca. e. respectively. Pierce et al. significance and large datasets: palisade layer adjacent to each epidermis and a spongy layer O’Leary (1981). the veins are surrounded by a distinct layer Hattersley and Watson (1992). subsequently rinse with tap water for 1–2 h and to discriminate declines. and are ‘instantaneous intrinsic WUE’. fresh samples from the comparative studies. CAM species show a distinctly lower pH after the night vapour pressure on transpiration rates. sections are an alternative way to keep records for later and respectively adjacent to. and contain smaller chloroplasts with no starch grains. precautions outlined in Section 3. is of great ecological interest and can be measured on do not have typical C3 palisade or spongy mesophyll layers.g. 1000 mL distilled water and C depends on ci.10. Therefore. it could be classified as (possible) CAM. If ci is low relative to ca. Pyankov et al. (1989). viz. gs is stomatal conductance. WUE is often containing cells just under their epidermis. Ehleringer (1991). it is growing conditions can have widely different C gain per likely to be C3 unless the plant is particularly succulent and unit water loss. In its simplest form. between the two palisades). This excludes the effect of differences in in the leaf. However. and possess abundant. (1998). The C-isotope approach has proved extremely useful to study A range of methods is available for making the microscope WUE over longer time scales.3 mL 10% H2SO4 for 2 weeks. chloroplast. Mohr and Schopfer (1995). Hibberd and Quick (2002). in contrast to many living. (1) Permanent slides or photographs and chloroplast visibility.. C-isotope ratios can share the same stomatal diffusion pathway. and environmental factors.4‰ and leaves. (1999). be aware that some may result in photosynthetic enzymes discriminate against the heavier stable poorer visibility of the chloroplasts. Because in a CAM plant. Farquhar et al. The extent of the enzyme’s discrimination against 13 alcohol (formaldehyde alcohol). and the ability of the enzyme for 1 week.10). Fig. For (see Section 3. such that different species in different If Kranz anatomy is observed. a = 4.12). thin-walled. Many CAM leaves WUE). On short time scales (= instantaneous). the ratio of the rates of net belongs to one of the families with CAM occurrence. In addition. but only a thin layer of more or less isodiametric. normally Sage (2001). C4 plants. ‘Kranz anatomy’. typically exhibit More on methods: Farquhar et al. so that C in the green colour of the chloroplasts is to soak the plant or leaves is always depleted in 13C compared with that in the leaves in a solution of 100 g CuSO4 in 25 mL of 40% formal atmosphere. The relative magnitude of photosynthesis and C3 leaf anatomy (see More on methods below in the present transpiration depends on several physiological. often enlarged chloroplasts that contain large starch granules. for C3 plants. In the latter photosynthesis and transpiration (= water use efficiency. the species is C4 If not. walled. Belea of bundle-sheath cells (Material S4. the upper to lower epidermis (see assessment. we recommend taking into account the same leaf population) at pre-dawn. turgid where D is photosynthetic 13C discrimination. contain no chloroplasts. Zotz and veins (transport structures with thin-walled phloem and generally Ziegler (1997). where A is net photosynthesis. This quantity. Earnshaw et al. short or long time scales. It relies on the fact that slides permanent. but with diffusion of provide further evidence to distinguish between CAM and C3 or water being 1. Material S4.6 times faster than that of CO2. ca and Special cases or extras ci are the mole fractions of CO2 in ambient air and in the substomatal cavity. usually organised into ‘palisade’ and ‘spongy’ layers parallel to. of the leaf consisting of large. then the air inside the 0. colourless parenchyma However. which can be sectioned free-hand by using a suitable b = 27‰. Fig. (1997).200 Australian Journal of Botany N.5). the ratio of net photosynthesis broken down during the day to supply CO2 for the photosynthesis to stomatal conductance. Ehleringer et al. One method for retaining isotope 13C (relative to 12C) during photosynthesis. (2000). with the entire centre measured with infrared gas analysis (see Section 3. morphological Section). This makes separating liquid obtained by crushing fresh leaf samples in the afternoon species effects and environmental effects challenging. Note that D is calculated from of several leaves. as follows: D = (d 13Cair – d 13Cplant)/ . material thus greater proportion of 13C than a plant performing photosynthesis treated can be sectioned only by using a microtome after at a higher ci. and again (with new. water-use efficiency These differences can usually be identified easily under an ordinary light microscope. and to calculate organic (mostly malic) acids build up during the night. an additional check could be to determine the pH of the intensity and vapour pressure deficit. typical vapour. however. The mesophyll cells are usually concentrated around the bundle-sheath cells. instantaneous WUE changes dramatically for a cells that store water and organic acids. thicker-walled xylem cells). 2) that are often thick- et al. often as a single layer whose cells are radially oriented relative to the centre of 3. D allows time-integrated estimates technique such as sectioning a rolled-up leaf or a stack of ci : ca and intrinsic WUE. the plant ends up fixing a store in 4% formal alcohol until use. case. then in 4% formal alcohol leaf becomes enriched in 13C. intrinsic WUE C4 metabolism (see (A) C-isotope analysis above in the present relates to the CO2 gradient as follows: Section). Pérez-Harguindeguy et al. If living plants are within given leaf over short time spans. Wand et al. 2) (vertically held C3 leaves often have a References on theory. (2002).13 C-isotope composition as a measure of intrinsic the vein. Intrinsic WUE ¼ A=gs ¼ ðca  ci Þ=1:6 ¼ ca ð1  ci =ca Þ=1:6. Many plant physiology and Uptake of CO2 through stomata inevitably leads to loss of water anatomy textbooks give further illustrations of Kranz v. embedding or freezing it. Lüttge (1997). because of variable light easy reach. As CO2 and water vapour than they do in the afternoon. called bundle sheath cells. Photomicrographs of freshly prepared d 13C (see Section 3. Taiz and Zeiger (2010). in contrast.

to minimise differences resulting from acclimation to different temperatures in the (1) Cellulose extracts. In other words. Note that. try to collect them Special cases or extras within the shortest possible time interval. Process the leaves on the day of the carbohydrates from snap-frozen leaves. e. The answer will depend on the question Samples should be dried as soon as possible and finely ground. Leaves at different eliminating a cell’s ability to retain solutes such as ions. dicotyledons). and changes in electrolyte conductivity of a solution bathing the also because of differences in the isotope composition of the tissue.14 Electrolyte leakage as an indicator of frost sensitivity What and how to collect? Electrolyte leakage after freezing is an indicator of leaf frost For intrinsic WUE assessment. (2010). being asked. 3. one of the first effects is disruption of membranes. Complete submergence is important. and is not affected by cuticle thickness. the bulk leaf 13C : 12C References on theory. Depending on the contrast of interest. In most cases. it is generally reasonable to assume that the isotope composition of air is constant and equal to that of the lower atmosphere (d 13Cair What and how to collect?  –8‰). however. so as to minimise natural senescence processes. For It is also important to note that. and that the equivalent in leaf fragments) in 1 mL of deionised water in each of equation for D given above is a simplification of the theory. the leaves from the point of the gradient where the species is most abundant. The technique described here is different ages. collection should be Grind the dried tissues thoroughly to pass through a 40-mm-mesh standardised across taxa. with mesophyll water on a shaker. significance and large datasets: ratio of fixed C correlates with the ci:ca ratio for the time period Farquhar et al. If many species are considered. the isotope composition of the air needs (from tender to sclerophyllous) and taxa (monocotyledons and to be known. not useful for estimating intrinsic WUE. then blot dry and submerge two disks (or their conductance being a particular complication. The technique is suitable for a wide range of leaf types source air. see below intrinsic WUE from C-isotope composition involves within the present Protocol).12 for measuring C-isotope concentrations. different isotope composition of other C compounds. Collect young. collect See Section 3. We reiterate here that the estimation of (to provide for two treatments using two disks each. fully expanded sun leaves with no sign of Storing and processing herbivory or pathogen damage. on tree rings geographical distribution. C-isotope composition is or measurements of the isotope composition of the air. Ion positions in a tree or in a canopy can vary in D as a result of leakage from a tissue can be easily assessed by measuring differences in stomatal opening and photosynthetic capacity. and in these groups. and also means that leaves that grew in based on the idea that when a cell or tissue experiences an acute different years or different seasons can have different D. however. Seibt et al. the 13C : 12C represents a (2008). avoiding the main veins. the equation given for D does not apply to C4 or many replicates (one replicate being two tubes.g.1). Collect leaves from at least five randomly chosen adult cellulose extracts to avoid variation introduced by the slightly individuals of each species. Rinse the samples for 2 h in deionised (photosynthesis to transpiration ratio). there is fractionation regions and/or growing at warmer sites along a steep regional between leaves and stems. because of their different each treatment (see below within the present Protocol). although in most cases. photosynthetic flux. has implications for the sampling strategy. Cernusak et al. Diefendorf et al. d13C is usually determined for sensitivity and is related to climate. each containing . during which the C comprising the leaf was fixed weighted by the More on methods: Ehleringer and Osmond (2000). Shorter-term (typically at the Storing and processing scale of a day) studies of D have sampled recent assimilates Store the leaf material in a cool container until processed in the rather than structural C.1). with a cork borer cut four circular 5-mm-diameter leaf disks (2) Assumptions. the D values of the whole tissue and those of cellulose correlate very well. which thermal stress. Deciding when to collect is more complicated. season and plant leaves. In freely circulating air such as at the top of a canopy. it is recommended to sample and or preferably near the end of the season (see Special cases and grind larger amounts of tissue to ensure representativeness. especially reflecting ci : ca during favourable periods. This enables of species from colder regions and/or growing at colder sites differentiation between growing conditions using tree rings of within a regional gradient. with all non-photosynthetic organs climatic gradient have shown greater frost sensitivity than those being more enriched in 13C than are leaves. or finer screen. (2009). To estimate D. C-isotope ratio analysis requires only small collect foliage during the peak growing season (see Section 3. Because intrinsic WUE changes rapidly. extras below in the present Section) or in winter (for winter evergreen species). (1989). and the objective is an interspecific comparison. either by extracting non-structural laboratory (see Section 3. If a species grows along a wide environmental Measuring gradient. Leaves of species from warmer for a historical record. two Eppendorf tubes. but can be determined on any plant part. in general. cut fragments of the photosynthetically active tissue necessarily correlate well with the actual WUE that add up to a similar LA. longer-term measure of ci : ca. prepare as biochemistry. that intrinsic WUE does not like leaves. or by sampling harvest. For each phloem sap.New handbook for measurement of plant traits Australian Journal of Botany 201 (1 + d 13Cplant). samples (2–5 mg). For needle- several assumptions. Isotopes are sometimes analysed using field. plant. which highlights the requirement for assumptions CAM plants.

To control 3. the For calculating the mean. . (6) Chilling sensitivity. tissues may tolerate physiological desiccation. Prior to immersion. units MPa) is a simple indicator of leaf water status. freezing treatment.  PEL in the control treatment: Alternatively. experimental manipulations and differences in injury when leaf disks or fragments are cut. to the two leaf disk/fragment samples in the recommend normally performing the procedure with leaves respective tubes: (1) incubation at 20C (or at ambient collected at or near the end of the growing season. or if one wishes to detect temperature and then measure the conductivity of the solution. (2002). More on methods: Earnshaw et al. let the samples reach ambient at about 8C is not available. pre-dawn YL may be more hemicryptophytes.202 Australian Journal of Botany N. cavitation in the xylem and osmolyte is not necessarily applicable to deciduous plants and accumulation in the cell walls. This tolerance tentative index of soil water availability. Alternatively. A chilling-sensitive tissue would leak electrolytes after such incubation. e. subtract the PEL of the control treatment of each replicate from that for the freezing Species facing soil water shortage can avoid water stress to a treatment. (5) Sensitivity to high temperatures. releasing all solutes into the external solution. Measurement (2) Season of collection. (1990). the average corrected PEL for each individual When measured pre-dawn. as it has been for leaves. Thus. Then place the Eppendorf tube in a could be detected using a freezer at 8C. and the YL may thus represent the soil water potential in the ‘average’ root Special cases and extras zone. or by shutting stomata Corrected PEL ¼ PEL in the freezing treatment and losing stored water only slowly through their cuticle. (2) Corrected PEL – the PEL of the control treatment can vary (2002). is a physiological limitation that can be Calculations ecologically important in mountains at lower latitudes. However. temperatures (~40C. in an ordinary frost treatment and the control for each individual plant refrigerator. temperature. and should be used only as a involves stems and buds rather than leaves. among species because of intrinsic differences in membrane permeability. The same basic technique. Gurvich et al. freeze plant tissue.g. then re-measure its conductivity. the more negative the value.15 Leaf water potential as a measure of water status for these and other sources of error. Incubations should be for 14 h in complete with the rest of the protocol the same as described above. Kyoto. namely and (2) incubation at 8C in a calibrated freezer. Earnshaw et al. to our knowledge. significance and large datasets: Levitt PEL indicate significant disruption of membranes. puncture with a modification in the treatment temperature. where es is the conductivity of a sample immediately after the treatment. Japan) and by (4) Acclimation. It is usually tested for by (1) Percentage of electrolyte leakage (PEL) – separately. the greater the frost sensitivity. because their significant frost tolerance negative than the soil water potential. A different treatment. It would not detect tolerance to merely tube into a standard previously calibrated conductivity meter mild frost. darkness. cell injury. The bulk leaf water potential (YL. because this will not actually PEL ¼ ðes =et Þ  100. has been the cap of each Eppendorf tube to allow relief of pressure during successfully applied to leaf sensitivity to unusually high boiling. Horiba. Corrected PEL is thus degree by dropping leaves. (such as the Horiba C-172. equilibrated with the soil during the night. without any prior occurrence of autumnal acclimation in frost tolerance. and thus (1980). membranes. although the reliability of this has not been investigated. Blum (1988). including technique is not suitable for halophytes and succulents. whereas a chilling-tolerant tissue should not. can be used if a freezer whose temperature can be controlled After applying the treatment. the plant may have become plant replicate counts as one statistical observation. 0C in a distilled water (or rain replication. for the incubation at about 78C (the temperature of dry ice). as stable as possible) for the control treatment. the higher the PEL. (1990). The occurrence of acclimation to mild frost recording the conductivity. as follows: water) bath could be used. we acclimation. to avoid light-induced reactions. see More on Methods below in the present Section). sampled. for a species. standard deviation or standard error more dehydrated the leaf. Gurvich et al. for the incubation for 24 h or more at about +5C. It nocturnal transpiration. Pérez-Harguindeguy et al. two disks or equivalent leaf fragments) as the number of plants could possibly be tested with sections cut from stems. Because of the recognised wide Apply the following two treatments. tolerance to the kind of severe frost that can occur at high Do this by placing a sample of the solution from an Eppendorf latitudes or altitudes. or delay the development of water stress in their tissues by rooting deeply. on leaf samples boiling water bath for 15 min to completely disrupt cell collected on successive dates in summer and autumn. and might be detected by this technique. High values of References on theory. The water potential as a result of several mechanisms. (3) Incubation with dry ice. and et is its conductivity after boiling. recent work has shown instances of substantial disequilibrium between pre-dawn YL and soil (1) Applicability to different plant functional types.

Samples should be collected at midday and. This consists of a pressure container into which the sample (leaf or terminal twig) is placed. the ‘balance by similar constraints (e. The simplest way to measure leaf water potential is with a References on methods: Scholander (1966). be kept refrigerated and in darkness (e. When the pressure between the first and last measurement. Minimum leaf samples cut out from fresh leaves of a whole range of species water potentials usually vary from near 0 to 5 MPa. Pressure. either leaves or short. Compressed gas cylinder What and how to collect Pressure Measurement of minimum values of YL is typically carried out gauge at the end of the hot. pressure chamber. Depending on the type of pressure chamber used (see below within the present Protocol). the midday YL can provide a useful index of the degree of physiological drought experienced. the balance exhaled to increase moisture and CO2 to try to minimise shoot pressure represents the equilibrium water potential of the plant material in the transpiration within the bag. These be lower in (semi-)arid ecosystems. However. (2006). Assuming that the xylem osmotic potential is very low. palatability tends to be correlated with the N tank. is then gradually increased in the chamber. or Scholander bomb (see diagram in Fig. with the cut end (with the number of samples depending on the number of pressure protruding from the seal. (2008).16 Leaf palatability as indicated by preference measure the pressure inside the chamber. the minimum value for YL that a plant reaches. Ackerly (2004). Extreme care should be taken experiments can provide useful information about herbivore when pressure chambers are under high pressures. Bucci et al. needle valve and pressure-safe (normally copper) tubing. Turner (1988). connected to the chamber through a Despite the vast diversity and complexity of herbivores. pressure is gradually applied from the gas cylinder. Bhaskar and Ackerly (2006).New handbook for measurement of plant traits Australian Journal of Botany 203 During the day. dry season for evergreen species and in Mediterranean winter-rain ecosystems. Thus. usually at midday at the driest. Assuming that the xylem osmotic potential is very low. from shoots or individuals located in the sun. Bartlett et al. or at least within half an hour of collecting all samples Fig. multiplied by –1. Samples should be in the chamber equals the xylem pressure. from traits. as indicated balance pressure represents the equilibrium water potential of by model herbivore preference. Jacobsen Measuring et al. leafy twigs should be collected. 4. YL will decline below the soil water potential Clean-cut surface Rubber gasket as a result of transpiration into the atmosphere. Samples sealed in plastic bags should chamber. and their interactions. When measured in the dry season. into which one has just surface. with its cut end can be seen as an integrator of several underlying leaf-quality projecting to the exterior through the sealing port. significance and large datasets: Hinckley et al. low nutrient contents. Leaves should have Chamber been exposed to direct sun for at least 30 min before collection (avoid cloudy days). as previously indicated (see Section 3.1). A cut branch (or leaf or compound leaf) is placed inside the chamber. the time of year at which drought stress is maximal may not be obvious. a drop of water appears at the cut collected into sealable plastic bags. high pressure’ indicated by the gauge or manometer is recorded. a manometer or pressure gauge to 3. When a litter decomposability among species because both are limited drop of water appears at the cut end of the specimen. Additionally. Repeated-measurements in different seasons can help find the real minimum YL for each species. or an insulated cooler box containing pre-frozen cooling bars or ice). (1978). Once the chamber has been sealed (hermetically chambers available) over a period of no more than half an hour closed). Lid hottest time of year. is a cafeteria assay in which the plant material in the chamber. the One method for quantifying leaf palatability. can be used as an index of the tolerance to water shortage that the species (or individuals and populations) demonstrate (assuming that the plants are still healthy and not drought-injured).g. concentration of lignin and secondary metabolites). in summer-rain ecosystems. Leaf water generalist herbivores are allowed to feed selectively on leaf potential is conventionally expressed in MPa. Measuring water potential with a pressure chamber. Leaf palatability (as indicated by preference by herbivores) A leaf or shoot is placed inside the chamber. (2012). References on theory. but can distributed in random positions on a feeding arena. preference for a broad range of plant species. Lenz et al. 4). by model herbivores a pressure tank of liquid N. . terminal. plants. (2004). We recommend measuring samples as soon as possible. way.g. a small number of components of leaf Many models with different characteristics are commercially quality affect preference by generalist herbivores in a predictable available. and as a pressure source. in a refrigerated picnic fridge.

1999). The %LA consumed can before the trial and stored in refrigerated and moist conditions. 1996. at least two cafeteria basis of the original cut shape.1) before and after the trial. Values of LA additional samples from a known material. plastic bags containing several consumption measurements can be compared with moistened paper towel) to preserve turgidity. (2003).) require a cool.204 Australian Journal of Botany N. by offering whole shoots with and without spines to different animals (in this case. It is recommended to assess 10-mm lengths from the mid-leaf section and pinning these down leaf palatability in at least two independent feeding trials. To be sure that the model herbivores do not have previous experience with Storing and processing the plants included in the trials. The surface is covered by transparent plastic in the case of trials with snails. placed. After herbivores have been placed. 1996). such as lettuce for consumption can be transformed into values of leaf biomass human consumption. What and how to collect environment. accessibility. (2000). accessibility. making sure common species of accurately (see Section 3. dark and humid environment Hartley and Jones (1997). Actual LA may also be measured trials should be carried out. on a numbered grid cell on a polystyrene feeding arena (Fig. provided contrasting quality are included in both for cross-calibration. either the animals or the feeding conditions are theoretical background on which palatability tests are based. Cornelissen et al. be estimated by eye (with 2–10% accuracy per sample) on the If different species grow in different seasons. (a) the arena can be constructed on polystyrene and (b) the numbered grid should be wrapped in plastic. For example. significance and large datasets: Grime position and the arena is closed with a plastic net (Fig. 5). without exposure to All leaves are kept in sealed bags at 4–5C until processed. For narrow leaves. together into a star shape. 5). Pérez-Harguindeguy et al.
Instead of selecting one recording time. 8 and 12 h. Snails are of highly succulent and aphyllous species. Coley (1987). several herbivores (~10 per 500 leaf samples) are placed in a random References on theory. Be aware that all species must be collected within 2 days subsequently every 12 h for 3 days. preferably from at least 10 individuals per is measured by direct observation after 4. any of the plants included in the cafeteria experiments. grasshoppers and photosynthetic tissue) is used as a leaf analogue. . Diagrammatic representation of a cafeteria (following Grime et al.g.1. probably not right. model herbivores Measuring should be bigger than snails or grasshoppers) and recording how much biomass is consumed per unit time. Tiny leaves may also need to be grouped using different model herbivores to cover a wider range of like this (with minimal overlap) to make up to ~1 cm2 In the case preferences by generalist herbivores. the herbivores may be bred. or collected when young and raised in captivity. Singer (spraying. securing with a thin needle. and species. Experiments can be designed behaviour. Snails et al. for these cafeterias (snails and slugs). Southwood et al. Grasshoppers need a dry and light. Large veins Special cases or extras should be avoided unless leaves are too tiny. and crickets a dry and dark More on methods: Pérez-Harguindeguy et al. consumption area in Section 3. Cornelissen et al. Once all samples have been pinned onto the arena. If the model herbivores are snails. This and a Once all leaves have been collected. dark plastic cover) that will stimulate consumption. If the animals do not eat from known preferred to evaluate palatability v. (c) Each leaf sample should be pricked on the grid. an equivalent area is reached by cutting an appropriate number of (1) Independent feeding trials. The arena should be covered with netting to avoid Leaves are selected and collected as indicated in Specific leaf any escapes. can be useful to test animal (2) Palatability v. (1986). water-saturated environment (e. 10 1-cm2 samples (one from pre-trial 48-h starvation period (promoting consumption during each individual) should be cut from each fresh leaf and randomly the trial) are important to get unbiased results. which is a substrate that snails like for crawling on. While crickets are better at discriminating between leaf qualities preparing all leaf samples be sure to keep the cut leaves in a within graminoids.1). following the same material. 5. Helix spp. (a) (c) 1 2 3 4 5 6 26 27 28 29 30 31 51 52 53 54 55 56 76 77 78 79 80 81 101 102 103 104 105 106 (b) 126 127 128 129 130 131 Fig.g. (1999). the samples do not deteriorate during this procedure. (1970. Including analyse-first choice and successive choices. but they of epidermis and adjacent mesophyll (relatively young consume few graminoid monocots. (e. avoiding the contact with the plastic so as to prevent rotting. a thin 1-cm2 fragment recommended for their generalist-feeding habits. popular with some of the herbivores used if SLA is included in the calculations (see Section 3.

P or base-cation content in others. as well as N and lignin (if not. with litterbags simply on subjectively judged to have died very recently and are still the soil surface. In leaves that gradually present Protocol). In species that shed their litter. (e. In particular community or several communities. particularly in litter beds. stapling (with non-rusting staples) or by using a Special cases or extras in the present Section). To complete senescence without any evidence of decay. however. the litter can be stored in the same bags for up to with demineralised or rain water before incubation will get several months in a relatively dry environment away from them to field capacity more quickly. Very big leaves may have to be cut into part) because it is an expression of the quality of litter as a substrate sections. it is recommended to divide the bed in equal statistical collecting several times from the same plants. Often such dead leaves are still shiny and. with one replicate of each species in a random position leaf litter. leaf pH. so as their litter. roots) on the quality of fibreglass mesh. Litterbags Here.g. we recommend beds. land use or natural succession are strong drivers morphologically contrasting species. litter must never be dried at high temperatures. For each species. In any case. and covered with Petioles and rachides of compound leaves that are shed as an the same litter mixture (Fig. made by wooden or undecomposed. collect those homogeneous across the bed. from the ground within a few days after falling or by placing a clean cloth or net beneath the tree or branch and gently shaking it Measuring until senesced leaves fall. stems and branches. it can be either collected inside.e. for microorganisms. Comparing some of the been to compare mass losses of (leaf) litters of multiple species by same species at different mesh sizes will help calibrate among incubating them simultaneously in semi-natural outdoor methods. narrow-leaved worldwide. 1 g) of dry litter. To preserve its and retrieved immediately after burial to control for loss or characteristics. soil before further processing. whereas wider mesh sizes allow a more CO2 To estimate the organ afterlife effects of different species complete set of organisms to be involved in decomposition and on decomposition rates. to prevent differences in leaves in which the middle lamina section is at the right phase. Finer mesh size should be of changing litter decomposition rates in various biomes used when. but litterbags should be made of non-degradable. Filling the bags using funnels or tubes inserted into the bags temporarily may help here. It is important to brush or rinse-off any Litterbags are incubated in a purpose-built outdoor litter bed. in plastic squares (with natural drainage) filled with soil thoroughly deciduous woody species. mesh type or and shape can be incubated during the whole period to check . to get representative blocks. with feedbacks to climate through the release of species are included. with representative proportions of midrib and petiole. better cut them into segments). i. Typical litterbags measure from through the ‘afterlife’ effects of attributes of living plant 10  10 cm to 20  20 cm and are made of 1-mm polyester or modules (leaves. the composition should be die from the tip down. we focus on leaf litter. mixed with a standard or combined litters. some reference species at two or three different initial masses.1). tannin. In species that retain dead leaves on Litter beds can range from just a rather homogeneous square in the plants or die back completely above ground. however. If there are large differences content. although the protocol can also be can be sealed with polycarbonate glue. a aphyllous species. Spraying the samples Once dry. undecomposed leaf litter should be collected from mature plants with code embossed) is firmly attached to the litterbag or sealed in the field. Plant species exert strong control over decomposition rates inert but flexible materials. or in some Species-specific ‘decomposability’ thus derived integrates environments. we recommend incubating related to SLA. Make sure a resistant label (e. Decomposability is usually related to leaf whereas flexible long monocot leaves can sometimes be rolled up dry-matter content. ingression of particles during burial. to reduce heterogeneity in Storing and processing moisture dynamics among the samples. Additional control Litter collections should be air-dried in open paper bags until samples of some of the species may be treated identically. where the entire shoot functions as the green more standardised mixture not based on local communities photosynthetic unit analogous to leaves and also senesces as a may be used for particular purposes (see below within the unit. filled with pieces of plastic broadly representing litter amount There are no fixed rules regarding litterbag size. Shifts in species composition. among the litters to be compared. for calibration among species afterwards. For species further account for environmental gradients within larger in which leaves senesce sequentially in cohorts. in each block. Additional litterbags calculate initial oven-dry weights of the samples for incubation. to a much more elaborate bed. additionally. for some species. may still show bright autumn colours. such units are collected as leaf litter. cleaned from vegetation and litter. It is important not to break What and how to collect the leaf litter during handling. The litterbags are usually incubated 1–2 cm below the surface of this mixture. plastic. a powerful method in recent years has have less effect on the litter microclimate. decomposition rate caused by the microenvironment. the composition of this mixture may be based on a processed as such (but see discussion under Section 3. they reach their equilibrium moisture content. so as to from the activities of mammals and birds. and again after 48 h in an oven (60C). A litterbag usually contains a standard small amount ‘common-garden experiments’. it may be necessary to increase or decrease the several structural and chemical traits of the leaf (or other plant initial mass per litterbag. as in many monocotyledons. it has been found to be in initial mass among some species. Freshly senesced.g.New handbook for measurement of plant traits Australian Journal of Botany 205 3. 6).17 Litter decomposability mesh material. Litterbag sizes may vary within one study. as a consequence of to standardise the ‘relative packing’ of litter inside the bags for global changes. Stretching 3–5-cm-mesh sunlight. leaf toughness. cut off leaves that are field. in most studies. by sewing (with nylon applied to other plant parts with some adjustments (see under thread). an air-dried subsample of leaf litter nylon net or a metal grid over the bed may protect the samples is weighed. laboratory heat-sealer. Depending on the type of integral component of the leaf litter should be collected and study.

Pérez-Harguindeguy et al. M0 = mass of litter quality. litter quality. samples may environments and periods vary greatly. 6. tweezers and small brushes (Fig. Decomposition beds and litterbag cleaning. Images of (a) decomposition bed fully covered with litterbags. second. After retrieval. In the absence of such (a) (b) (c) unprocessed sample sample processing Fig. so that mass loss or k either be cleaned up and processed directly. as follows: according to the microclimatic conditions that will determine decomposition rates and.206 Australian Journal of Botany N. then weighed. or first frozen. involving more than one harvest. almost 90% mass loss can be reached in less than 2 months. Mt = mass of litter at time t and t = duration of the minimum time needed for over 50% mass loss for the same incubation (years).g. litters within litterbags. to the average quality of the k ¼ Ln ðM t =M 0 Þ=t. We advocate the establishment of one or a few be removed from the decomposed litter. given high where k = decomposition rate constant (year–1). whereas in arid or cold environments. in incubation period or as k (decomposition constant) derived a controlled environment greenhouse. Two retrieval dates are usually enough to check for the consistency of decomposability ranking across species over Calibration among studies time. could provide close to rate can be defined as the percentage of mass loss after the standardised environmental conditions from year to year. whereas five or more may be needed if decomposition A disadvantage of the litter-bed assay is that the incubation dynamics are to be analysed too. 6). or then oven-dried for 48 h at 60C. Retrieval dates and sampling schedule may vary. It would be better still if such litter beds. values can be compared directly only within but not between Adhering soil. Decomposition equivalent ‘common garden’ facilities. for the contribution of exogeneous organic matter to the from a presumed exponential mass-loss curve over time sample. (b) litterbags covered with natural mixed litter and protected by a wire mesh and (c) incubated-litterbag cleaning with brush.g. 2 years may be litter at time 0. soil fauna and other extraneous materials must studies or sites. e. In moist tropical environments. first. . to the degree possible centralised litter beds. which could simultaneously host samples using e. Litter samples are from multiple sites.

(2009). In combination with plant size-related traits. are also of the branches. defence. Soft-wooded or herbaceous samples are more References on theory. SSD is emerging as a core sites. 2007). because of biomechanical and fauna or leaching is studied. functional trait because of its importance for the stability. microcosms can be sampled it also plays an important global role in the storage of C. but see Special cases or extras in the stem is placed within the water. the slice should be 2–10 cm thick. we recommend (1) additional inclusion of a few species density’ in that SSD can also be measured on herbaceous from each range of sites in a multispecies litter bed. Cornelissen in plastic bags. Garnier et al. or else cut off are coordinated between organs across species. hydraulics. fine and coarse stems). This procedure allows the volume of irregularly shaped samples. What and how to collect? chemical changes and biological colonisation.3). and the extent to which these If possible. (1) Microcosms. cut with a knife or saw a ~10-cm-long (4) Litter decomposabilities of other organs (fine and coarse section near the base of the stem (between 10. so a representative sample free mass (%) of the initial and final litter samples (this is not must include a proportional representation of the complete stem. stems (diameter <6 cm). (2008).1 Stem-specific density enough to hold the entire sample. As physical damage. (2007). remain constant from pith to bark. for affect the species rankings of mass loss in incubations under large trees. at certain time intervals during the process of decomposition. the volume of the same section.3-m include coarse woody debris together with fine litter in such height. Stem density can increase. and tare the balance. and lower for branches and dry or windy environments with high irradiance. One should be aware that in some (because of tapering of vessels). is filled with distilled Stem-specific density (SSD. or when the researcher needs to hydraulic safety. suggestions. Cadisch and Giller (1997). Freschet et al. a low stem density (with large vessels) leads to a fast growth. C gain and growth potential Special cases or extras of plants.New handbook for measurement of plant traits Australian Journal of Botany 207 facility. A new method is available to diameters >6 cm. Graça et al. when still fresh. so as to be able species and includes the stem bark (i. If contamination is too high and difficult to only the loose bark that appears functionally detached from the eliminate. Parton et al. (2004). Fortunel et al. branchless section. roots. so should be wrapped in moist tissue Cornelissen (1996). decrease or 450C. (2005). . appropriate for soils high in organic matter). resistance against pathogens. Hard-wooded samples comparative common-garden studies. (2012). placed on a balance. Although most litter beds are situated under because of cheap volumetric construction costs and a large field conditions. a triangular sector from the bark tapering into the exposed conditions or below a litter layer. significance and large datasets: vulnerable to shrinkage. herbivores or standardise the incubation conditions for any other reason. and the mass loss expressed on the basis of the ash. For woody or thick succulent plants with stem great ecological relevance. (2012). remove or clay fraction. mass loss for outer bark (which includes more air spaces and cork). the sample density or wood density should be measured (see Special cases or can be oven-dried and then sieved to remove bulk of the sand extras below in the present Section). taking into account can be stored in a sealed plastic bag (preferably cool) until differences in size and time scale of decomposition. large 4. When sand or clay contamination is high. A graded test tube or beaker. This trait is a The wood sample is then completely submerged under the synonym for ‘stem density’. Adair et al. and stored in a cool box or refrigerator until et al. either stem even after removing obvious extraneous matters. select a regular. saw out a (pizza) slice from the trunk at ~1. secondary phloem and to calibrate mass-loss values across multiple sites. architecture. Freschet et al. Measuring More on methods: Taylor and Parkinson (1988). Species differences centre (like a pizza slice) of cross-sectional area. Collect either a whole stem cross-sectional slice or. to be sure that when the mass (at 70C for 72 h. Berg and Stem volume can be determined in. litterbags can also be incubated in hydraulic capacity. Austin and Vivanco measurement. Stem density partly underlies the growth-survival trade- off. (1999. Healthy adult plants should be selected according to previous (2) Contamination. Cornwell et al. Laskowski (2005). and the incubated material can be analysed for its mass loss. which in some cases accounts for a significant across multiple species from these sites or (2) inclusion of litter proportion of the overall stem structure (see Special cases and samples of a few reference species in multiple litter beds across extras in the present Section). the incubated litter sample can be ashed for 4 h at stem (see Section 4. in common-garden experiments. mg mm–3 or kg dm–3) is the oven-dry water (but not completely filled. For herbaceous species or woody species with thin main of interest. and thereby cork if any). we distinguish it from ‘wood water with the aid of a small-volume needle or tweezers.and 40-cm height). Cornelissen (1996). measurement.e. to be measured easily. Stem density is higher at the insertion point of branches (3) Photodegradation. For stem density. Robertson et al. (2006). (2008). (1999). at least. Depending on the objectives of the study. whereas a high stem density (with small microcosms when more detailed effects of soil and litter vessels) leads to a high survival. These resulting from photodegradation or sand abrasion (in addition density gradients have important implications for the sampling to microbial decomposition) may be substantial and this may procedure. which cannot be 4 Stem traits properly measured with the dimensional method. two different ways. (1) Water-displacement method. ~1/8 of the total in photodegradation at a given exposure may themselves be area. the liquid will not escape present Section) of a section of the main stem of a plant divided by from the test tube).

most of the structural before multiplying by L. Try to isolate the short. in turn.800 ADW + 0. until a constant weight is obtained. this stem again usually has stored in a plastic drinking straw. In some plants. can safely be used as SSD. containing substantial bound being registered by the balance (i. Samples should be wood density (oven-dry wood density of the main trunk dried in a well ventilated oven at 101–105C for 24–72 h (small including sapwood and heartwood) from xylem density or samples). Thus. If the stem is tissue. the ‘wood density’ is often measured at leaves. however. rope can be tied around the tree and fastened at the handle of because bark usually makes up only a very small fraction of the the borer. or a reduced. Many trees V ¼ ð0:5DÞ2  p  L and shrubs in savannah-type vegetation (and some Mediterranean and arid species) have. Pérez-Harguindeguy et al. make separate density estimates for wood and bark. which. Because wood is increase on water level leads to an increase in the weight mainly cellulose and lignin. which equals the wood sample volume molecular weight.e. and density is reported as ‘air-dry weight’ (see Section 2. oven. and often the some bamboo species). SSD as described in the present protocol is called ‘stemless’ is probably more convenient from the point of ‘oven dry weight’ (ODW). using callipers. such a sample may not be leaves are attached to an underground. previously replacing water. Because a core does not taper are attached. rope will wrap itself around the borer. which (4) Xylem density. before determining both registered in the balance (i. stem has. all the inward towards the centre of the stem. directly measured or random one if they are all similar. either no. the density of whose stem can ground (‘breast height’). Besides free water. This method can be applied to samples support is given by the wood and wood is generally denser having different geometrical forms. They then refer to the relationship sample) is quickly read and registered. The considered to be air or water spaces that do not belong to volume of a cylindrical sample can be determined simply by the stem tissue. measurement.e. very thin. because Calculate the volume (V) of the cylinder as they have very different chemical and cellular compositions. . perfectly representative of the density of the stem as a whole. for example. increment borer. whereas smaller spaces such as the lumens of measuring its total length (L). a very In the case of hollow stems (e. samples are often taken with an plants. stems also contain bound water. the (1) Oven drying and wood-specific gravity. and its diameter (D). or a densities). After volume measurement than bark. terminal need more drying time. a the contribution of the bark of a tree to its stem density. with the ends of the straw sealed. young trees of Cecropia or thick corky bark (with very low density). Tare again for each between weight and volume as wood specific gravity. C exchange and growth. branches of trees). physical properties and biological functions. using a calibrated ocular micrometer.g. After the core is extracted. the weight of the water and relatively small quantities of compounds of low displaced water). Some authors make a distinction between is removed only by drying at above 100C. condensed stem. many shrubs).99) stem density as zero. In these species. When coring trees with very dense wood. select the apparent main branch.3). qualitatively recording it as volume). because density is not weight. estimate the diameter of the hollow and volume of the bark may be an important part (even 50%) subtract its cross-sectional area from the stem cross-sectional area of the stem volume. If a plant branches from ground level (this formula cannot be correct for ‘dry weights’. to which the leaves to just beyond the centre of the stem. increasing the tension except as noted below. derived from ADW. and obtain its density. Most rosette plants and basal-leaved graminoids Samples with this method are usually taken at ~1. near ground level. determine the diameter on a cross-section of it (3) Wood density v. and intercellular spaces.g. it can be also be determined. often horizontal. ~1. are part of the stem more places along the sample. Very large holes in the stem are (2) Measurement of dimensions (or dimensional method). stem density. This value ignores (6) Dense woods. function in supporting photosynthetic In the timber industry. on the rope.5-cm. but weight/ ground leaf-supporting. The mass dry wood samples at 100–110C.3 m above produce aerial inflorescences. the sample is dried in the the important parameter. the difference from a density estimate using a sector or density can be determined.3). Additional useful methods from forestry (5) Plants without a prominent above-ground stem (rosette In forest ecological studies. Large-diameter of mechanical leaf-supporting function that an above-ground corers (12 mm) are better because they cause less compaction. (2) Stems with holes. the volume of the wood weight and volume. than quantitatively recording its by using the formula ODW = 0. however. which helps push the borer bit into the tree.0134 (R2 = 0. but which does not have the kind an entire stem section is usually probably small. whose however. many wood scientists and foresters oven- in cm3 (because water has a density of 1 g cm–3). It might be worthwhile to under a microscope.208 Australian Journal of Botany N. this error is probably unimportant. in which a wood core is cut from the bark inward. If the plant has no recognisable above- (ADW) (a misnomer. Xylem density has been proposed as a proxy for the tree hydraulic architecture. compared with that of an extensive-stemmed plant 12% moisture content. from the support viewpoint. wood density is (by any of the methods described above). modified stem called a rhizome (see Section 2. Large samples may sapwood density (measured on small. on one or xylem vessels. When the handle is turned around during coring. grasses and sedges). We suggest that data for ODW. ADW can be transformed to ODW view of further analyses. being careful to not touch the sides or bottom of the beaker Special cases or extras which can cause variations in the weight being registered by the balance. When the wood sample is submerged. the mass of a large tree trunk. stem. may limit tree performance in terms of transpiration. but only for (e.

adult plants should be sampled as indicated above paper and put them in sealed plastic bags. compared with that in Section 4. (2008). water paper. Following the rehydration procedure (see Section 3. on the diameter of the trunk Collect one to three terminal (highest ramification-order. If no cool box is available in the field and samples that are used for measurements of SSD (see Section 4.1).1). Santiago et al. in that case. In general.1. the procedure can be combined with that for relevance in trees or large shrubs subject to surface-fire regimes. or lower. Loehle (1988). it includes the vascular cambium. other phenols. on the position of bud primordia within the bark or smallest diameter-class). Twig drying time is expressed in days the main stem of forbs or the leaf sheaths of grasses. Twigs (or twig sections) should preferably be provide protection of vital tissues against attack by pathogens. where 100% is the final loss of mass when the this number may involve an error because the density of weight of the sample ceases to decline further at the indicated bark cell-wall material is not necessarily the same as that temperature. dry-matter content are expected to dry out relatively quickly during the dry season in fire-prone regions. a ‘main twig’ with fine side twigs can be collected as one suberin in cork. (1992).411  branch wood density. using if necessary). Williamson and Wiemann (2010).1). Swenson and Enquist (2008). adopted here. (2001b). Gartner produce in twigs a dry condition relatively similar to that from air- et al. or branch. so Measuring using it can sometimes introduce an error. Measure bark thickness on a minimum of five box or fridge (never in a freezer!). lignin. the equivalent Twig dry-matter content (TDMC) is the oven-dry mass (mg) of a to TDMC is stem dry-matter content. which is defined negatively correlated with potential RGR. number of days it takes to reach 95% of the mass reduction of wall material (1. this trait has special stem. water. Twigs with high More on methods: Garnier et al. Every 24 h. (2004). Shipley and Vu (2002). the sample as a result of drying (interpolating between weightings if an appreciable fraction of the stem volume is bark. Special cases or extras 4. SSD (see Section 4.2 Twig dry-matter content and twig drying time (i) Herbaceous plants. Continue until you are certain that a steady dry for wood. sample. Thick bark may also of five plants. Brown (1997). take the main herbivores. We consider TDMC to be a critical References on theory and significance: Bond and Van Wilgen component of plant potential flammability.New handbook for measurement of plant traits Australian Journal of Botany 209 (7) Measuring other than main stem. 20–30 cm long. Reyes et al. Main-stem wood density procedure once back in the laboratory. The volumetric fractions of cell-wall material. significance and large datasets: Putz weight obtained at 40C is not the true dry mass of the twig. then follow the above 1–2 cm in diameter may be sampled. measured in exactly the same way as LDMC but using expressed in mg g–1. the rather low drying temperature Anten and Schieving (2010). although this relationship can vary among species. (1983). although the effectiveness depends on the intensity and duration of a fire. (1992).1). Patiño et al.3 Bark thickness (and bark quality) positively correlated with specific density or dry-matter content of the main stem across woody species (see Section 4. drying outdoors. is chosen so as to More on methods: Reyes et al. For herbaceous plants. conductivity after ignition (see Section 2. TDMC should be 4. Lavorel and Garnier (2002). Store these in a cool (see Section 1. divided by the original volume (in cm3) of the 40% relative humidity of the outside air. because some bound water will remain within the cell walls of Gartner (1995). Each twig sample (consisting of 1–3 volume fraction of water is simply the decrease in weight (in g) twigs) is then first dried in an oven or drying room at 40C at on drying. often important components of bark defence as well.1). If wood samples cannot be temperatures are high. (until equilibrium moisture).53 g cm–3 for cell-wall material in dry wood).g. to remove any surface water before measuring water- and gas (‘air’) in the stem can be calculated as follows. the material (and probably also in its protein) at this temperature Poorter et al. For very fine. not been tested explicitly. and Bark thickness is the thickness of the bark in mm. preferably (to minimise damage) on the same the laboratory. hence. Van Gelder et al. The volume fraction that is cell-wall material equals each sample is reweighed. gums. (2009) (see Section 4. until further processing in adult individuals. (2004). although this has. it is better to store the samples in plastic removed from a trunk or main stem. has been found on average to equal 1.3). any (8) Components of stem volume (cell-wall material. Thick bark insulates meristems and bud primordia from lethally high temperatures What and how to collect? associated with fire. (2006). but which can be obtained relatively quickly. leaves are removed and the twigs gently blotted dry with tissue gas). However. tannins. . Storing and processing What and how to collect? Wrap the twigs (including any attached leaves) in moist Healthy. (2009). wood from branches bags without any additional moisture. twigs. divided by its water-saturated fresh mass (g). The saturated fresh mass. (2005). If a plant has no branches or twigs. which can be terminal twig. particularly fire (1996). The final dry References on theory. Chave et al. frost or drought. The fraction of the original volume that was gas is weight has been reached. TDMC is defined (analogously to simply 1 minus the foregoing two volume fractions. strongly ramifying terminal Be aware that the structure and biochemistry of the bark (e. Twig drying time is defined as the the dry mass : fresh volume divided by the density of dry cell.12). resins) are unit. Chave et al. et al. to here as the part of the stem that is external to the wood or xylem – our knowledge. Niklas (1994). LDMC) as dry mass divided by saturated mass. sun-exposed twigs from a minimum cambium and on bark quality and moisture.

(2012). the rate of forestry). and smaller branches or twigs. KS is a function of the numbers and conductivity of individual xylem conduits and their interconnections via pit (1) Bark quality. cork cambium or cork). KS typically measurements of both the maximum (outside the fissure) declines towards the leaves because of tapering of vessel and minimum (inside the fissure) bark thickness. greatly across species. If you do not use the same sample as for SSD. Take the average per sample. Special cases or extras in the present Section). secondary phloem. Bark services. and. provided they are kept We suggest five broad (subjective) categories. In the simplest case. see Point 1 of Special cases or extras in the present (3) Fissured stems. has advantages (more related to fire resistance) and problems Measuring (often the base of the tree is deformed).5 mm). because that is where surface fires occur (but see Brando et al. It can also be quantified thickness are made with callipers (or special tools used in as the LA-specific xylem hydraulic conductivity (KL). The complementary measurement of (use filtered. for which the methods are simplest. in forestry water-transport system. 40-cm height. 10–30 cm. take five random Section). off. several membranes. Measure this trait on main stem near the base between 10. been devised for herbaceous plants. In situ measurement with a purpose-designed forestry tool is relative to the transpiration demand of the foliage that the branch an acceptable alternative. including cool and with their ends in water. Avoid warts. An alternative can These methods have been developed largely for the stems of be to make this measurement at ~50–60 cm (and in any case woody plants. Xylem refers here to the conducting tissue in the stem thickness (mm) is the average of all sample means.and References on theory and significance: Jackson et al. if possible to the nearest 0. In most cases. The bark. five random measurements of bark gradient of pressure (kg m–1 s–1 MPa–1). considered the minimum conductivity for the stem part of the (4) Alternative height for measurements. so that the measurement includes the resistivity of both the (amplitudes 0. breast height) or directly at DBH.5 mm). nutrients and organic matter. in many cases. whereas in some angiosperms. (1999). bark thickness is measured at breast height (as DBH). and after known time intervals. Measuring at the base of the tree. At the investments (by dividing the bark thickness by the stem distal end of the segment.8). KL equals KS. Conduit length varies 2–5 mm) and very coarse texture (amplitudes >0. (3) intermediate texture xylem. wood (i. Then numbers and sizes. a collecting container catches emerging radius). as suggested here. specific objectives may ordinary laboratory equipment can be used. Segments ~30 cm long are commonly used. in cylindrical tubes).4 Xylem conductivity few centimetres wide and long. Decorticating bark is usually for performing xylem-conductivity measurements have been considered as standing litter. cut out a new piece of bark of at least a 4. degassed water) through a stem with a known cross- stem diameter can be useful to compare species for bark sectional area and no (or with sealed-off) side branches. leaves and roots. stem samples brought in from the field. Pérez-Harguindeguy et al. known pressure head is applied to push a 10 mM KCl solution (6) Bark investment. The efficiency of water transport is quantified as the stem-specific xylem hydraulic conductivity (KS).1 mm. because these may differ vessel length can reach more than a metre (although it is usually greatly and support different epiphyte communities. described. and so they can be them. The tested branches should be (1) smooth texture. the rate How to measure? of water flow per unit cross-sectional xylem area and per unit For each sample or tree. a imply its measurement). see Special cases or extras in the present Section. so it is not included in bark.210 Australian Journal of Botany N. In each sample. divided by the ratio between LA and sapwood cross-sectional area (see Special cases or extras Section 2.5–2 mm). (4) strong texture (amplitudes conduits and the inter-conduit connections. its volume is determined .e. includes everything external to the By replacing the water lost to the atmosphere via transpiration. liquid. as defined here. KL expresses the conductivity of a branch. phelloderm water transport prevents highly negative and damaging leaf or secondary cortex. however. for trees. gymnosperms having short tracheids Bark textures may be measured separately for the trunk (usually less than 1 cm long). What and how to collect (2) Bark surface structure (texture) may determine the capture The measurement may conveniently be made indoors on and/or storage of water. Relatively sophisticated types of apparatus (5) Decorticating bark. any vascular cambium. water potentials from developing. however. measurements are made on calculate bark thickness as one-half the difference between stems collected from the outer canopy. For species with water flow per unit of supported LA per unit pressure gradient fissured stems. lumen diameter (as can be modelled using Poiseuille’s law of flow An easy but possibly important one is the presence (1) v. Typically. surveys. In addition to bark thickness. equates to sapwood. Analogous bark thickness at 50 cm is strongly related to bark thickness at methods have. simple systems built from thickness measurements (however. absence (0) of visible (liquid or viscose) gums or resins in the bark. (kg m–1 s–1 MPa–1). and permits continued photosynthesis. (2) very slight texture (amplitudes of longer than the length of the longest conducting element in the microrelief within 0. Conductivity structural or chemical components of bark quality may be of an individual conduit increases with the fourth power of its of particular interest (see above within the present Protocol). thorns or other protuberances and remove any bark pieces that have mostly flaked Water transport from soil to leaves is critical to land-plant activity. per unit of xylem cross-sectional area.

However. on the x-axis. Sperry (2003). Tyree and horizontally (and symmetrically) across the top of a Zimmermann (2002). Section 3. values of xylem conductance. allowing a conductance measurement flow through that element. on Special cases or extras secondary branches that are removed at intervals for this purpose. conductance of a (1) Length of vessels. Zimmermann and Jeje (1981). (2004). partially dehydrate. on the y-axis. North and Noble can be inferred. The xylem vulnerability to embolism indicates the risk of loss of Details needed for actually employing this method are given water transport during drought. at curve by one number. effectively blocking water completely through it. its length (in m) divided by 10 should be added to xylem vulnerability curve. This simple method is based on the principle given stem-water potentials. force that was applied. and develop tension as a result of transpiration from its leaves. In the laboratory. Alder et al. (1) Evaporative dehydration. To characterise this tree trunks. and its conductance is details on the procedure). This affords a value for the percentage loss of that remains is likely to be a little longer than the length of its conductance at that particular Y. or ones that cannot be reliably attached to an available rotor. or longer. L is the length of the segment (m). Proceed as indicated in Fig. If the stem segment is held vertically during the Vulnerability to embolism is quantified by constructing a measurement. Sperry et al. cut several stems in the field (see Fig. 7. Vulnerability is expressed as the in some of the cited references. water within the xylem conduits. centrifuge rotor and spinning it generates tension in the (2004). Brodribb and Hill (2000). . similarly tested to obtain conductance-loss values for other References on theory. Sack and segments can be brought to different effective water Holbrook (2006). (2001). which is not easy to determine for Stem segments with different Y values can be obtained by any routine measurements.15) and is an important aspect of drought tolerance. which is applied to a stem segment that is times the atmospheric pressure. the sapwood area. This is needed to ensure that the length of segment of the main axis near the last-removed lateral is stem segments used for conductance measurements exceeds measured. the average of the of xylem conductivity. Cavender-Bares et al. which can in some cases become as high as 100 pressure. Conductivity is then calculated tolerate highly negative water potentials (= high tensions) without as embolising varies greatly among species (cf. however. K S ¼ J  L  A1  DY1 . Any air entry into a water. A large branch that includes to transport water under a unit pressure difference. percentage of the water-saturated xylem conductance that is lost at (3) Air injection. This consists of plotting measured the applied pressure (MPa) (but not if the segment is horizontal). Advantages of the method are that stem (1992). one can usually these conductance values (represented as a percentage of the assume that A is the entire xylem cross-sectional area (all of the maximum water-saturated conductance) were obtained. (1988). (2001). 7 for further from it (see references cited below). (2008a). When a target Y has been reached. the value of Y at its mid-point (50% a maximum. by the pressure chamber method (cf. the while the external pressure is applied. for its xylem to and L terms. Maherali et al. Kocacinar and Sage (2003). a corresponding (negative) Y or P References on methods: Sperry et al. (1996). however. Separate branches are longest vessel. Section 3. often an even smaller loss of conductance) is commonly used. The more such emboli develop. areas at the two ends of the segment). A would have to be the conducting xylem area. of three possible methods.New handbook for measurement of plant traits Australian Journal of Botany 211 either directly or gravimetrically. Xylem conductance refers to the capacity of a vascular system.15). the technique is difficult for stem segments longer than ~20 cm. Zwieniecki et al. potentials very quickly. Y is determined periodically. from the upper re-measured to obtain the maximum conductance of the end of one of these. and it cannot be used 4. against the KS is typically reported in kg m–1 s–1 MPa–1 With measurements values of stem water potential (Y).. (2004). at which on terminal or subterminal branch segments.5 Vulnerability to embolism for segments longer than the width of the rotor chamber. A is the mean cross-sectional area of the Follow the above instructions for collecting for measurement xylem of the stem (m2) (to a first approximation. Santiago et al. significance and large datasets: values of Y. remove a portion such that the segment segment. with larger branches or shape of this curve is usually sigmoid. The procedure uses a conducting element will dissipate the tension in it and quickly special pressure chamber designed for a stem segment to pass expand (becoming an air embolism). located within a pressure chamber. The water stream in the conduits is of substituting internal tension by external positive air under tension. From the centrifugal Holbrook and Zwieniecki (2005). Conductance some lateral secondary branches is cut from the plant and can be calculated from the above equation by simply omitting its A kept unwrapped during the day. and DY is the pressure difference (MPa) between the upstream and downstream ends of Measuring the segment. with whatever length and cross-sectional area it happens to have. area is actually conductive. (2) Centrifugation. The xylem being conductive). Water is then flushed through this segment briefly that of their vessels. Attaching a short stem segment Meinzer et al. What and how to collect where J is the rate of water flow through the stem (kg s–1). This type of chamber greater the loss of xylem conductance. The ability of a species to is commercially available from the PMS Instrument Co. To determine the maximum length of the under a pressure high enough to displace all air embolisms vessels.

If bubbles emerge from that end. From this segment. References on theory. however. fine roots selectively acquire difference in measuring just SRL or its two components. Procedure to determine the vessel length. The segment’s length at this point is just shorter than the length of its longest vessel. Sperry et al. the stem’s longest vessels are shorter than that segment’s length. Oregon. significance and large datasets: Tyree and Sperry (1989). while modifying soil chemistry via exudation of a functional traits of roots. accessed help better understand the below-ground strategies of multiple 15 February 2013). whereas roots with low resources between below-ground and above-ground parts. (2008a). Albany. Roots are often measured in aggregate when comparing the There is no single evolutionary solution to all the variable live fine roots of plants. With the syringe’s plunger a mild. Then a segment slightly longer that this (but one that does not emit bubbles when first tested) can be used to come close to the maximum vessel length as at the end of the foregoing procedure. we mineral nutrients from a complex range of soil solutions and recommend that both metrics be measured when measuring the soil particles.1 Specific root length Maherali et al. (1997). Pammenter and Vander resource investment (mass). References on methods: Cochard et al. Yet. so relatively long pieces must be cut successively from it until bubbles do emerge.1). If. (2002). at least one vessel has been cut at both ends of the segment. proceed as described below). Fine roots are the primary organs for water and with different traits. (2003). Feild and Balun (2008). high SRL can result from having a low diameter Variation in root traits among species has large ramifications for or low tissue density. numbers of fine roots can be enough to measure root diameter or Measuring several independent root traits in combination may branching order. although individual fine roots or small challenges of acquiring soil resources in different ecosystems. Separating fine roots according to the timing . Plants with high SRL build more Willigen (1998). Repeat this test using progressively longer stem segments until one is found from which bubbles do not emerge under pressure. the ratio of root length to dry mass of et al. When under high nutrient conditions. (1992). shorter root lifespan and higher RGRs than for low- SRL plants. Fine tissue density have lower longevity but greater rates of uptake roots can acquire resources directly or through symbionts. Sperry et al. Davis et al. (2004). retest the segment. after each cut. no bubbles emerge from the first stem segment that is tested. and a relatively large (e. (2007). cut off successively shorter slices and. 50 mL) air-filled syringe is attached to the free end of this tubing. tight-fitting diameter is slipped over the segment’s debarked end. allowing air to move freely through the segment’s entire length (if no bubbles emerge. range of compounds. Pérez-Harguindeguy et al. providing a ratio of a standard unit of acquisition (root length) to (1988. Alder et al. until bubbles are first seen to emerge. (a) A segment of approximately the probable length of the given species vessels is cut from the stem of interest and bark is removed from its basal end. Here. each of which is independently associated their ecology. is the below-ground analogue of SLA (see Section 3. 2008b). Roots also must respond to heterogeneity of resource availability at multiple spatial and temporal scales. fine roots. thin roots exert less penetrative nutrient acquisition and they are also responsible for transferring force on soil and transport less water. Holbrook and Zwieniecki (2005). 5.212 Australian Journal of Botany N. USA (http://pmsinstrument. we describe three main sets of traits. Choat Specific root length (SRL). (a) (b) rubber tubing receptacle air under pressure with water with syringe bubbles stem debarked emerging segment end stem stem debarked end Fig.g. species. while resisting attacks from a wide range of organisms and What and how to collect? environmental stresses. Brodribb et al. As there is little operational accessing soil resources directly. root length for a given dry-mass investment and are generally considered to have higher rates of nutrient and water uptake (per 5 Below-ground traits dry mass). For example. above-atmospheric air pressure is generated within the tubing while holding the segment’s free end under water. (b) A rubber tubing of a suitable.

and plucking of debris with forceps to diameter and tissue density have similar cellular structure. glass slide. roots morphology of fine roots. diameter and tissue density requires digitising the roots and Craine (2009). Craine et al. or preferably slightly cellulose purple or red–violet. cell diameters and amounts of different tissues can be generally have a lighter. A scanner that has the top 20 cm are the standard basis of comparison. floppy or deflated. fine roots should be traced which also aids in teasing individual roots apart. the a transparency adaptor that illuminates items on the scanner bed actual depth sampled should be allowed to be as varied as the from above. Böhm Once roots have been obtained and prepared. Quebec. units of root length need to be measurement generally fits in the palm of your hand. it is often most at 60C) and weighed. atypically Section 3. Canada). root dry mass. A useful rule of thumb is to in a polymer. In mixed-species assemblages. If each. A clear plastic back to shoots for positive identification. Live roots data. Washing the software. scanned roots should be dried (48 h a small number of species. Nevertheless. and radius). A typical amount of root necessary for When roots have been scanned. Roots measured in the field span a range of root age. healthy roots from the recently washed sample. analysed for nutrient concentrations. These root samples can also be ground and feasible to excavate the entire plant to be washed out later. which stains lignin blue–green and the fine roots as you are removing soil. Fitter (1996). Pregitzer et al. There should be no need to stain most roots to uniform stands and roots can often be distinguished among image them. we also recommend cross- can be ashed at 650C later and ash mass subtracted from gross sectioning roots. order. is recommended to provide crisp root images. Digitisation can be carried questions. It will help to observe a range of ages References on theory. require hours of painstaking plucking. For small plants. (2002). Roumet et al. Paula and Pausas (2011). so as to determine their cellular structure. diameter and Storing and processing root volume distribution of a sample of root length. roots are often divided by root (ramification) (2009). multiple roots of each species are embedded solution for longer periods of time. Washed roots can be stored in a 50% ethanol For this purpose. With these live. Bouma et al. Often roots have to be finger-massaged and (mainly phloem and xylem) as well as the construction of individual roots separated to allow particles to be removed. More on methods: Newman (1966). For a small number of is better to have a small amount of root that is better prepared than roots. . under a dissecting microscope. remove contaminants that are of a similar size and density as the Roots can vary in their relative proportions of cortex and stele roots of interest. with little degradation of structure. endodermis and large xylem elements are determined by tracing each portion of If necessary. however. The basis of comparison should be clear and still half the width of the finest roots of any plant. (root dry mass over volume. A scanner with a whereas roots acquired from ingrowth cores or young plants resolution of 1600 dpi provides a resolution of 15 mm. WinRhizo (Régent Instruments. whereas more rigorous washing might be more use. For a large number of samples or large or small individuals should be avoided. out with almost any low-end flatbed scanner. so as to properly identify healthy live roots. cleaning roots will require a combination of running water over a fine mesh sieve (0. After scanning. This is not necessary in tray works well. the latter being derived from length Washing techniques should be gentle for species with low. For these reasons. The software will automatically determine the length. saved suitable for high-density roots in soils with heavy clays or coarse in the JPEG format and analysed later by someone who owns organic matter that could compromise measurements. (2006). Preferably. average root diameter and root tissue density conditions for a week. the commercially available application. In general. Tennant (1975). and then mounted on a before. Craine woody species. stele. fully turgid appearance. Section for manual methods when none of the above facilities is whereas clearing roots of organic matter from a tundra soil might available. compared with calculated relative to one another and to total cross-sectional dead or dying roots of the same species which appear darker and area. In general. a root acquisition and preparation considered each time.2–1 mm) to remove fine heavy particles such as Special cases or extras sand. sort apparently the root manually in image-analysis software. (2000). significance and large datasets: and colours of absorptive roots for each plant species before Eissenstat and Yanai (1997). SRL. it traced and their diameter determined. root length. determining (1979). to better standardise comparisons across species. See under Special cases and extras in the present roots from a sandy soil can require as little as 30 s under a hose. Steudle measurement. Not all roots of a given particles such as pebbles. aiding in identification. Although the software is expensive for occasional density roots. Roots height above ground from which to collect leaves. which is would constrain age. are best imaged while submerged in a small amount of water. cut on a microtome into 4-mm slices. stained stop washing roots when it appears that you are losing as much of with toluidine blue. Roots from scanner with lower resolution may also work.New handbook for measurement of plant traits Australian Journal of Botany 213 or depth of sampling could be informative to answer particular measuring their length and diameter. secondary to measuring gross some fine particles such as clays are too difficult to remove. roots can be scanned independent of analysis software. Digital images are made for each species using light microscopy at 100 magnification and the cross- Measuring sectional areas of the root. is recommended. Wahl and Ryser (2000). (2001). however. note that live and dead roots may not be distinguishable by colour. enabling easy Unwashed roots can generally be stored under humid. rinsing in containers of water to remove coarser heavy (1) Root diameter and tissue density.1 for free software). this can be carried out with image-analysis software (see a larger amount of less well prepared root. For (2001). chilled calculations of SRL.

those root sections that define their vertical distribution of influence on soil activity. consider adding a dispersal depth. Furthermore. as exemplified by the recent extent. and/or submerged in large tubs. agent (e. To determine We provide specific protocols in Material S2 that deal with the lateral extent of root systems. and intensity of exploration. whereas understanding root length with depth is likely to best kept separate from the relatively thin sections. it is simpler to determine the root biomass with different or resource storage. Pérez-Harguindeguy et al. In are obviously particularly important for mechanical support general. below individuals for species with a tap root. 5. Hodge (2004). If the soil is particularly clayey. usually exceeding 10 mm in diameter. Root systems can be affecting this is SRL (see Section 5. Jackson (1999). In other cases. Caldwell and Virginia species. Linder et al. or would be strongly correlated if there were no change in SRL with contains calcium carbonate. It also determines the dried and weighed for biomass distributions. many plants exploit symbiotic associations relationship. and the ability of a species to compete for soil nutrients. Schenk and rooted species. Confirming the identity of individual roots and need to be measured explicitly. In general. . Withington et al. (1989). Lambers et al. an auger of differences in the affinities of the ion-transport carrier and in the 5–10-cm diameter should be used to remove biomass with depth. The best washing additive varies depending on the related to longevity and negatively related to nutrient uptake. Zwieniecki et al.2 Root-system morphology laid out on a large mesh screen. Subsamples of ranging from a few centimetres to tens of metres). They can be a better metric to understand the competitive ability of uptake still be combined for certain analyses later on. However. If depth distributions are to be determined. it is most easily carried out by lateral extent. capacities of the roots among different species are superimposed Typical depth distributions follow a somewhat exponential on this. For more deep-rooted species. Craine Collection and analysis (2009). distance. Excavating entire plants is reasonable for some shallow. sodium hexametaphosphate) to the washing Note that root tissue density and root diameter are positively water. a horizontal strip of soil can be following strategies: excavated. Dunbabin et al. biomass and length with depth (2) Clayey soil. When sampling larger shrubs and The depth distribution of roots combines depth and intensity of trees. root tissue density is positively related to resistance to References on theory and significance: Adiku et al. fine (maximum soil depth from which resources can be acquired. to be washed out with running Characteristics of entire root systems can be independent of water. (<2 mm) and coarse roots should be separated. aggregated. if desired. or at a point that represents the midpoint of its lateral of species in less studied regions.1). 40. The maximum cleaned fine roots can then be scanned. (1) Large shrubs and trees. Within the coarse root fraction. with bacteria or fungi to enhance their nutrient-competitive 80. Regardless. 20. There are species might require anatomical or molecular comparisons with three main traits of root systems that are best measured: depth. and a cross-section of the soil from a pit face excavated. quantitative removed in entirety or in sections. Depth distributions are a better indicator of the best way to deal with this is to use a specialised wood-cutting relative reliance of plants on different depths for soil resources and auger. are depths. (2) arbuscular mycorrhizae – symbiosis with arbuscular Once soils have been excavated. 10. ability of plants to interact with spatial heterogeneity in soil resources. For rooting depth. In other cases. Incomplete root-depth distributions can be used to Symbioses with N2-fixing bacteria or mycorrhizal fungi have estimate maximum rooting depths. root biomass should be plant that roots can acquire resources from. 2 m of soil. Then interspecific differences in nutrient-uptake capacity. other roots of that species. which is the subject of the present Section. root storage and washing use mycorrhizal fungi to aid in acquisition of nutrients and the same protocols as described above. roots have to be accessed from caves or boreholes. for example. (2000). Schenk and Jackson (2002). however. In particular condition of the soil.214 Australian Journal of Botany N. literature is strongly on the pattern of root biomass with depth. although this should be verified. for diameter. tracing roots back to their above-ground parts or sampling in the simplest metric to determine is maximum rooting depth conspecific stands. The utilisation of soil. A standard set of depths would be 5. lateral extent of roots defines the distance from the centre of the length and volume analyses. 5. a deep pit is dug and one pit face is smoothed vertically. (2000). pathogens and drought. however. the researcher will encounter thicker woody roots. (2004). addition. depth distributions can be determined randomly relative to the There is still much to be learned about nutrient-uptake strategies individual. however. Intact root systems are best water. this will depend been found in many plant species. (2006). (2011). Determining the maximum extent of roots depends on the More on methods: Böhm (1979). With ecosystems is undoubtedly affected by their inherent pits.3 Nutrient-uptake strategy Depth distributions can be determined by digging pits if a The performance of different species in nutrient-limited known cross-sectional area can be excavated with depth. starting at the centre of the plant. One factor a cross-section is removed with a flat shovel. so as to trace roots outward. For some species. whereas biomass will have to be determined directly discovery of specialised N-foraging snow roots in the Caucasus. where individuals are bunched. (2002). capacity. The amount of fine-root biomass or root length per Special cases or extras unit soil volume indicate the intensity of soil exploration. a pit must be dug Jackson (2002). 120 and 200 cm for root systems largely confined to the top ability. biased towards temperate species of Europe and North America. In some extreme cases. the lateral (1) N2-fixing bacteria – association with bacteria in nodules to extent of roots is likely to be equivalent to half the interplant fix atmospheric N2.g.

g. the routes it can travel and its final nutrients directly from a host plant. Record all categories that are sedges and capillaroid roots in rushes. bills. thick-walled. Impatiens). ecto-endo-mycorrhizae (certain References on theory and significance: see Material S2. It is important to realise that dispersules may (occasionally) get (d) trichomes – nutrient and water capture through transported by one of the above modes even though they have bromeliad leaves. (3) Internal animal transport (endo-zoochory). (6) Ant dispersal (myrmecochory). Howe and Westley (1997). e. that extract nutrients such as N and P from The mode of dispersal of the ‘dispersule’ (or propagule = unit of the roots or stems of a host plant. often brightly coloured berries. many fleshy. in gymnosperms) and pyroloid mycorrhizae (Pyrolaceae). Van der Pijl (1982). where the whole plant or infructescence with ripe seeds is rolled over the ground by wind force. as seen in many shrubs and trees (e. by birds.g. wind dispersal takes priority over ant (b) baskets – nutrient and water capture and storage. (Arbutus.g. and (e. Acer. fungi to aid in uptake of organic forms of N.g.g. the seed or fruit has no obvious aids for longer-distance transport and merely falls passively from the plant.1 Dispersal syndrome (8) root or stem-hemiparasitic green plants. poplars (Populus). (7) Dispersal by water (hydrochory). Arctostaphylos). dispersal. (7) myco-heterotrophic plants without chlorophyll that extract C and probably most nutrients from dead organic matter via 6 Regenerative traits mycorrhizal fungi.g. addition. Baptisia lanceolata in the south-eastern USA and Anastatica hierochuntica (rose-of-Jericho) in northern Africa and the Middle East. drupes and big fruits (often brightly coloured). many grasses). leaves. References on theory. transport and use by ants or related insects. How to classify? (11) hairy root clusters (proteoid roots). fruits or seeds that become attached e. dauciform roots in This is a categorical trait. burdock (Arctium). Dispersal syndromes (1) Unassisted dispersal. ‘balloons’ or comas (trichomes at the end of a seed). e. spores of ferns and related vascular cryptogams (Pteridophyta) and (D) ‘tumbleweeds’. birch (Betula). dispersules with elaiosomes (specialised nutritious appendages) that make them attractive for capture. (c) ant nests – nutrient uptake and storage. that aid in P uptake. awns. (2009). feathers. hygroscopic bristles on the dispersule that promote movement with varying humidity. for snow roots. Poschlod et al. in the case of certain ferns with very thin fronds).g.New handbook for measurement of plant traits Australian Journal of Botany 215 (3) ecto-mycorrhizae – symbiosis with ecto-mycorrhizal fungi Although most of the experimental work on the active uptake to aid in uptake of inorganic nutrients and organic forms of of nutrient ions by roots has been carried out on agricultural N. assumed to give significant potential dispersal (see Box 4). The radioisotope techniques for characterisation of ion-transport (5) orchids – symbiosis with orchid mycorrhizal fungi for mechanisms are beyond the scope of the present handbook. arbutoid mycorrhizae literature. and no obvious adaptation for it. Tough. This is particularly true for endo- (e) root velamen radiculum – nutrient and water capture zoochory and exo-zoochory. (10) carnivorous plants that capture organic forms of N and P from animals. (9) Bristle contraction. bats. Hulme (1998). mammals. e. (4) ericoid-mycorrhizae – symbiosis with ericoid mycorrhizal but again relatively little for tropical and subtropical species. indehiscent nuts tend to be hoarded by mammals (e. destination. barbs.g. Bakker et al. lime (Tilia). dispersules are adapted to prolonged floating on the water surface. such as mistletoes (Loranthaceae).g. many Asteraceae). (1996). In the case of similar potential (a) tank plants (ponds) – nutrient and water capture and contributions.ort P-uptake mechanism. apple (Malus)).g. significance and large datasets: Howe uptake presumably directly through root hairs (or through and Smallwood (1982).) by jays). Orchidaceae). prioritise the one with the presumed longer- storage. to animal hairs. e. wingless seeds or nuts by birds (e. that are evidently eaten by vertebrates and pass through the gut before the seeds enter the soil elsewhere (e. restrained seeds that are launched away from the plant by ‘explosion’ as soon as the seed capsule opens (e. see Onipchenko et al. pine (Pinus)). (B) seeds with pappus or other long hairs (e. holly (Ilex). (2) Wind dispersal (anemochory) includes (A) minute dust-like seeds (e.g. in Floras) for dispersal mode of many plant taxa. plants. but can be readily ascertained by consulting the ion-transport (6) rarer types of mycorrhizae. aided for instance by corky tissues and low specific gravity (e. thereby distributing the seeds. fruit or spore as it is dispersed) has obvious consequences for (9) holoparasitic plants without chlorophyll that extract C and the distances it can cover. willows (Salix). Box 4.g. e. ash (Fraxinus). distance dispersal. Pyrola. some information on that of wild plants is available. (13) none – no obvious specialised N. (2000). . brown or green seeds or nuts that are hoarded and buried by mammals or birds. Note that there is ample literature and storage. aided by appendages such as hooks. in (12) other specialised strategies (mostly in epiphytes). including order of decreasing importance. seed.g. acorns (Quercus spp. (4) External animal transport (exo-zoochory). The latter strategy is known from arid regions. coconut). arillate seeds. hazelnuts (Corylus) by squirrels) and rounded. elm (Ulmus). (5) Dispersal by hoarding. (8) Dispersal by launching (ballistichory).g.g. legs. 6. acquiring nutrients from litter. burs or sticky substances (e.g. (C) flattened fruits or seeds with large ‘wings’.

significance and large datasets: Hendry Wenny (2005). which of the dispersule. most simply. highest standardised value for each dimension (length. (2004). after each of these values has been divided by the can be chosen arbitrarily depending on the question. because the characters may be correlated with dispersal potential. e. (above-ground!) canopy seeds banks air. such as e. Pérez-Harguindeguy et al.g. air-dry storage is also recorded either by direct measurements in the field or can okay. off between dispersal potential (in space) and maximum plant it may be efficient to pay local people specialised in tree climbing lifespan as well as seed-bank persistence (dispersal in time). pappus or other loose parts (see above dispersules and animal-dispersal potential to either attachment in the present Section). Releasing height We recommend complementing this trait with other direct or should be measured during dispersule release and is the difference indirect assessment of banks of seeds or seedlings for future between the highest elevation of the seed or fruit and the base of regeneration of a species. the higher the probability that one they have been through an animal gut system first). it constitutes the seed plus surrounding structures. be identified by measurements of traits related to the dispersal potential. Peco 6. however. Leishman and Westoby (1998). For naturally dry dispersules. as do species with a long-term persistent seed bank.e. it is a crucial variable when asking if dispersal is limiting the occurrence of What and how to collect? a species in suitable habitats or species richness of plant The same type of individuals as for leaf traits and plant height communities.3 Dispersal potential is its oven-dry mass. The dispersule may (1993. Weiher et al. Dispersule size 6.2 Dispersule size and shape et al. i. structure or propagule) as it enters the soil.g.216 Australian Journal of Botany N. Therefore. The seeds seed spans larger distances. The dispersules can form and structure of seed surface responsible for a high dispersal either be picked off the plant or be collected from the soil surface. 1997). Funes et al.e. (1993. Dispersule shape is the variance of its three Dispersal potential is defined as the proportion of dispersules dimensions. and Grime (1993). attached after a defined time) is measured by putting seeds on . Storing and processing How to record? Store the dispersules in sealed plastic bags and keep in a cool box or fridge until measurement. Hakea and Protea) and long-lived that are placed on a flask shaker moving with a frequency of seedling banks of woody species in the shaded understorey of 100 min–1. Of interest is the entire reproductive dispersule (= dispersal More on methods: Hendry and Grime (1993). mass. (or dispersules) should be mature and alive. Pons and Pausas (2007). Tackenberg et al. Floating capacity (proportion of dispersules floating after a of serotinous species of fire-prone ecosystems (e. seed or fruit with all the structures.g. e. Pinus and defined time) is measured by putting dispersules in glass beakers Proteaceae such as Banksia. assessments. (and identification) to help with the collecting. Weiher et al. 1997). The flesh of fleshy fruits is removed too. with the same dispersal syndrome. For seed-bank assessment. but also strongly among species seed persistence in the soil (seed-bank persistence). Dispersal by humans. (2003). There may be a trade In some parts of the world. the width and the thickness (breadth) produced by one individual that travelz a certain distance. (1999). dispersal potential allows the spherical) tend to be buried deeper into the soil and live longer assessment of dispersability of a seed in relation to distance. The seeds are usually the units to get buried in this case (certainly if more seeds are produced. Therefore. For the remaining dispersule. i. however. therefore. Thompson et al. or if fragmentation is a threat to the survival of should be sampled.g. there are the plant. (1999). wings and awns remain high dispersal potential. the fruit. It in the seed bank. 48 h) and weigh (= dispersule size). Measuring dispersal thickness) using callipers or a binocular microscope and calculate potential. Myers Vivipary as in some mangroves could also be part included in such et al. by the actual rate of fall in still present Section). (2003). dispersal potential by water to buoyancy of the Remove any fruit flesh. The largest of the three values. (1999). In unitless. requires studies adapted to the specific the variance. Seed size and shape are then fundamental for varies not only among species. pappus and Special cases or extras awns. in many species.g. Process and measure as soon Dispersal potential is a continuous variable and may be as possible. Long-lived species often exhibit a low dispersal potential. Then dry at 60C for at least 72 h (or else at 80C for question. potential depend on the dispersal vector. take the potential or survival after digestion. Wind-dispersal potential Measuring is correlated with dispersule-releasing height and terminal velocity. The seed characters such as e. Attachment capacity (proportion of dispersules still woodlands and forests may also make important contributions. in some tropical rain forest areas. that are still attached when it is released.g. Measurements should be carried out on the intact dispersule. Pons and Pausas (2007). Thompson et al. only parts that fall off easily (e.g. the length. Forget and References on theory. Small dispersules with low shape values (relatively contrast to dispersal syndrome. Of interest is the unit that is likely to enter the species or populations. correspond with the seed. width and machines or vehicles is very complex. whereas parts such as e. McIntyre and Lavorel (2001). pappus) are habitats or in fragmented landscapes is often correlated with a removed. Both seed production and also seed attached. The capacity to survive in disturbed soil. Variances lie between 0 and 1 and are dispersules may be seeds or fruits or vegetative propagules. More on methods: Howe and Westley (1997). Terminal velocity is measured on freshly collected air- good methods to follow (see More on methods below in the dry dispersules and. or by modelling approaches.

Westoby (1998). those data can be also The same type of individuals as for leaf traits and plant height successfully used. (2) Secondary process. equilibrium mass in very large or hard-skinned seeds) and weigh. Only leave the fruit intact in cases where the testa and the surrounding fruit structure are virtually inseparable.g. fruit flesh). e. Be aware that seed size may 6. Survival after digestion is measured either by digestion If dispersule shape is also measured. For certain (e. (2003).g. (2000). do not remove any parts mass may be useful in such cases. 2005b. in some tropical rain forest Wright et al.1). selection of species by grazing animals) strongly influences dispersal potential. Westoby plant may be needed for species with tiny seeds (e. whether or not wrapped in moist paper (see Section ingestion by a standardised mechanical treatment and digestion 3. more closely related taxa being more (3) Seed volume. (2005). To assess animal-dispersal potential. assuming an ellipsoidal shape). e. identification). seed volume. Predicting animal-dispersal After measurements of dispersule shape (if applicable). comas.e. Interspecific variation in seed mass also has an important used is compatible. Stored resources in from each individual. predict over a range of scenarios.g. potential requires process-based models with the ability to remove any accessories (wings.e. Thompson et al. Schurr et al. 2005) Tackenberg (2003). standard Tackenberg et al. deep (2) Available databases. (2007). field studies should be added where possible. accuracy of the balance available. healthy adult plants that have their foliage (4) Additional measurements. Most of these databases What and how to collect? actually include both seed mass and volume. then store cool in sealed experiments with the respective animals or by simulating plastic bags. Römermann et al. (2005). dispersal by wind on the ground. Poschlod et al. We recommend collecting at least 10 seeds from each of 10 plants of a species.g. If the shape of the dispersal unit (e. fruit) is structure) may be of additional interest. however. Both dry and fresh measured too (see Section 6. areas.4 Seed mass vary more within an individual than among individuals of the Seed mass.2 above). Dry the seeds (1) For water plants. until dispersule measurement is finished. References on theory. Special cases or extras (1) Within individual variation. et al. Moles et al. i. as one statistical observation for calculations of mean. allometric) exposed to full sunlight (or otherwise plants with the strongest questions. expressed in mg. and larger numbers with the same reproductive effort. (1998. drought. If they cannot be weighed immediately after often obvious only from field studies and may require the cooling down. Cousens et al. Seiwa and Kikuzawa (1996). first try to define clearly which parts belong to the fruit as a whole and which belong strictly to the Special cases or extras seed. single-seeded fruits) at 80C for at least 48 h (or until between the highest point of seeds or fruits and water surface. Depending on the Mazer (1989). The seeds should be mature dispersule unit or the entire infructescence (reproductive and alive. significance and large datasets: although more plants per species is preferred. In other words. it may be efficient to work in collaboration with local More on methods: Hendry and Grime (1993). seed. Moles and Westoby (2006). but make sure not to remove the testa in the process. put them in the desiccator until weighing.g. additional measurements of the mass of the light exposure for that species). unpublished databases may be accessible also tend to be buried deeper in the soil. make sure the methodology banks. Will and deviation and standard error. also called seed size. (1999). (2002). which have to be calibrated dry storage is also appropriate. Tackenberg (2008). by digestion experiments.New handbook for measurement of plant traits Australian Journal of Botany 217 the respective animal fur. should be sampled. particularly if their under certain conditions. seed releasing height is the distance (or achenes. Otherwise air- by a standardised chemical treatment. which is then shaken by a shaking Storing and processing machine. Make sure to collect ‘average-sized’ seeds average seed of a species. or else establishment of additional new methods. establish in the face of environmental hazards (e.g. pappus. elaiosomes. which aids their longevity in seed added to the database. the samples will take up may strongly affect dispersal potential. . taxonomic component. 2005c). and process and measure as soon as possible. Smaller seeds can be produced in of published data are already available in the literature. often measured as p/6  L1  L2  L3 (i. Reich et al. is the oven-dry mass of an same species. Be aware that a considerable amount shade. Smaller seeds some of the large. In some parts of the world. once taken from the oven. More on methods: Fischer et al. (2007). people specialised in tree climbing to help with collecting (and (1997). orchids). and not the exceptionally small or large large seeds tend to help the young seedling to survive and ones. Wright et al. (1996). Cornelissen (1999). There are also many large datasets for likely to be similar in seed mass. Be aware that. Many of these data can probably be shape is close to spherical. back in the oven to dry off again. (2010). Note that the average number of References on theory and significance: Bruun and Poschlod seeds from one plant (whether based on five or 1000 seeds) counts (2006). Using the appropriate calibration equations. because the behaviour of animals Measuring (e. Leishman et al. Weiher et al. 100 or even 1000 seeds per (1998). herbivory). Such processes are moisture from the air. (2005a.g.

whereas small seed sizes (1) cryptocotylar hypogeal with reserve storage cotyledons are related to foliaceous and photosynthetic cotyledons. Zanne et al. Kitajima (1996).218 Australian Journal of Botany N. PEF cotyledon position – epigeal when hypocotyl develops at least 2 cm above soil surface. Garwood (1996). the occurrence and proportion of each type in other (3) phanerocotylar epigeal with foliaceous cotyledons (PEF). disturbances. possibilities in relation to cotyledon exposure (phanerocotylar or The distribution of seedling traits across families is still rather cryptocotylar). seeds in black). pathogen damage and predation must be selected among the collected ones to run the experiments. Ibarra-Manriquez et al. created on the basis of woody species (trees and shrubs) and cryptocotylar foliaceous seedling types are biologically not has been mainly used in tropical forests. 8): related to reserve storage seedling types. (2001).6 Resprouting capacity after major disturbance Fig. Leck et al. however. PER = phanerocotylar epigeal with storage its persistence in ecosystems where recurrent major disturbances cotyledons. extreme drought or frost events. see below within the present systematics was recognised quite early. they are still considered reserve organs (e. (2005). In many cases fleshy CHR CER cotyledons do contain chlorophyll. and phanerocotylar hypogeal foliaceous seedlings have established the following five seedling categories based on not yet been reported. Because (CHR). e. (4) phanerocotylar epigeal with reserve storage cotyledons (PER) and What and how to collect? Because this is not a plastic trait. Garwood (1996) possible. cryptocotylar if cotyledons remain within the seed coat. 6. Aspidosperma spp. the above-mentioned types have been particularly identified for (2) cryptocotylar epigeal with reserve storage cotyledons (CER). Seedling functional types. we recommend germinating sufficient number of seeds to obtain about five seedlings per species. or cotyledon function – reserve cotyledon are fleshy. large seed sizes are (position. foliaceous cotyledons (also called paracotyledons) are primarily photosynthetic. Although eight combinations are potentially possible. severe . relation to cotyledon function and position. Species are then assigned to the seedling morphological categories indicated above in the present Section (and in Fig. It is a categorical trait These categories result from the combination of different that can be used to characterise plant regenerative strategies. cotyledons in grey. although the importance of seedling traits in (foliaceous or reserve storage. (2008). Seedling functional types are correlated three cotyledon characters of presumed ecological significance with other plant traits such as seed size. hypogeal when hypocotyl develops on the soil surface. References on theory.g.5 Seedling functional morphology (5) phanerocotylar hypogeal with reserve storage cotyledons Seedling functional types refer to morphology of seedlings in (PHR). for example. however. hurricane-force CHR = cryptocotylar hypogeal with storage cotyledons. 8): cotyledon exposure – phanerocotylar if the seed coat opens and two cotyledons emerge from seed.4 (leaves in white. Hladik and Miquel (1990). PHR PER Measuring Seedling morphology (cotyledon exposure. Pérez-Harguindeguy et al. Wright et al. PEF = phanerocotylar epigeal with foliaceous cotyledons. De Vogel (1980). Special cases or extras (1) Chlorophyll in fleshy cotyledons. This trait has been Protocol). position (epigeal or hypogeal) and function poorly known. 6. 8. Seeds without evidence of. significance and large datasets: Ng (1978).). ecosystems should be tested. (2000). are common. Five seedling functional types as The capacity of a plant species to resprout after destruction of described in Section 6. texture and exposure) (Fig. and CER = wind and logging are the most obvious and widespread major cryptocotylar epigeal with storage cotyledons. Fire (natural or anthropogenic). Seedling functional morphology: PHR = phanerocotylar hypogeal most of its above-ground biomass is an important attribute for with storage cotyledons.g. tropical forests. position and function) should be described when at least five individuals have developed at least three leaves each.

if a tree is still or size limits for resprouting ability may reveal important standing after a fire. cambium and young xylem insights into population dynamics. resprouting may be investigated experimentally by destruction could also be applied to the study of resprouting clipping plants to simulate destruction of 75–100% of the in the face of disturbances other than fire. direct comparisons reproductive output lower. and their species suffer the same disturbance regime. species composition tends to be associated with the likelihood of major biomass-destruction events. It is particularly relevant for resprouting. Be aware that basal or below-ground parts) and divide by 100 to obtain the species highly adapted to fire (such as these examples) may ‘resprouting capacity’ (range 0–100. Del Tredici (2001). In species where no resprouting is observed merely and Midgley (2001). the assessment may be carried out up to 5 years (2) Strongly clonal plants. However. after the disturbance (as long as shoots emerging from near the soil it is important to assess whether damaged ramets can resprout surface can still be identified unambiguously as sprouts following from below-ground reserves and not from the foliage of a biomass destruction). intraspecific error of up to 25 units as a result of the dependence of sprouters tend to show a larger allocation of carbohydrates to resprouting capacity on the severity of disturbance encountered below-ground organs (or storage organs at soil-surface level). dead xylem (wood) is be seen as a component of recruitment. However. substantial resprouting. rangers). should provide the evidence for that. can resprout from buds located in high that has resprouted (i. the clipped parts can be trashing by vertebrates. and 100. This is to ensure that regrowth is only supported by resprouting ability of young plants may reveal important reserves from basal or below-ground organs. If fewer than five plants with ‘appropriate’ damage can be found. old. In the case of strongly clonal plants. following a fire. Estimate (crudely) the average percentage of (4) Resprouting after smaller biomass destruction. 20. search for spots with clear symptoms of a which the more quantitative assessment is not feasible. Note that in the case insights into population persistence. In longer-term approach of recording resprouting after less severe biomass studies. Pausas (1997). this event should have e. it should be recorded as destruction of 100% of resprouting species cannot resprout before a certain age the above-ground biomass. (2000). it is flooding and other short-term large-scale erosion events important to consider this a missing value (not a value of zero). give the species References on theory. browsing or trashing by large herbivores. Additional recording of green canopy. because the non-resprouting individuals are hard to been within the same year. significance and large datasets: Noble a default value of 50 if any resprouting is observed (50 being and Slatyer (1980). 60. used for other trait measurements as well). live above-ground biomass was destroyed. Thus. as is common in some herbs. Special cases or extras (1) Data from literature. using reserves from basal or below- resprouting capacity after major disturbance as follows: 0. For each species. in such species. When data are give the false impression that an area has not been exposed to available from more than one site. The contribution of sprouters to should be safe. moderate between general applicability and rapid assessment on the one resprouting. also qualify. because no major biomass destruction can be found. or direct species value. including the entire (3) Resprouting of young plants. such as herbivory or above-ground biomass (in which case. 40. The variability in sprouting capacity may occur. 80.g. or size. Quercus suber and many percentage by the percentage of the damaged plant population Eucalyptus spp. for each species.g. landslides. abundant hand and precision on the other. The woody plants and graminoids. same crude estimates may also be used for species for Within the study site. Multiply this disturbances. Other species in the same area. foresters. It is known that some have been killed. In general. we define resprouting capacity as the relative ability of conditions of major destruction of above-ground biomass a plant species to form new shoots after destruction of most of have been met.e. Make sure that the same Here. Therefore. For instance. Bellingham and Sparrow (2000). within ecosystems where different however.New handbook for measurement of plant traits Australian Journal of Botany 219 grazing. very abundant resprouting. The following method is a clear compromise never resprouting. assign subjective numbers for its above-ground biomass. unitless). Compared with non-sprouters. but may also be applied to forbs. data on the age not considered as part of the live biomass. see Kammesheidt (1999). Useful and legitimate data may be How to assess? obtained from the literature or by talking to local people (e. resprouting. as well as to the degree of stress in terms of available resources. their biomass growth tends to be slower. but all its bark. There appear to be ecological trade-offs between Broad interspecific comparisons have to take into account an sprouters and non-sprouter plants. Additional above-ground biomass destroyed among these plants (a recording of resprouting after less severe biomass destruction measure of disturbance severity) by comparing against average may provide useful insights into plant response to undamaged adult plants of the same species. recent major disturbance event. Everham and Brokaw (1996). being considered. very poor resprouting. resprouting adult plants between 5 and 50 individuals (depending on time should be recorded only if most above-ground biomass available) from which as much as possible but at least 75% of the has been destroyed for all ramets in the vicinity. if only woody species are find after disturbance. farmers. and others may lose their resprouting capacity Make sure that enough time has lapsed for possible when they attain a certain age or size. ground plant parts. try to find any number of connected ramet. halfway between ‘modest’ and ‘substantial’ resprouting. Thus. Higgins et al. In such cases. although this could also of trunks and branches of woody plants. although this ignores the fact that great intraspecific fire observations. . formed new shoots emerging from positions along the stem. Bond below). take the highest value as the severe fires recently.

Publishers: The Hague. Finally. Korhola A. . hypothesis. 665–674. Domenach A-M. 25–44. Vose JM. in xeric and mesic habitats. Ecological Monographs 74. Aerts R. Sperry JS.plantsci. Functional Ecology 24. we thank plant species Aranwela N.’ 2nd edn.1365-2435. doi:10. 87–105. Pockman WT (1996) Root and stem xylem embolism.1469-8137. Aubert S. Cereal Research Communications 26. Schieving F (2010) The role of wood mass density and Adrienne Nicotra. (Eds MJA Werger. Burrows (2002). Nature 442.x re-evaluation of processes and patterns.1111/j. 9–44 (SPB Academic and belowground biomass allocation patterns and competitive ability. Vivanco L (2006) Plant litter decomposition in a semi-arid architecture of Pinus palustris maintain similar stomatal conductance ecosystem controlled by photodegradation. Roggy J-C. doi:10. doi:10. Starmer KP (2002) Leaf Nature Geoscience 3. American improved the quality of the manuscript. Bekker RM. Zaehle S. Pausas and Albert CH.1007/BF00320419 Bartlett M.’ 2nd edn.1461-0248. The Allen SE (1989) ‘Chemical analysis of ecological material.1046/j. New Phytologist 144. CONICET (PIP 2006-2011 11220080101532). Scoffoni C. Tjoelker MG (2005) The hot and the cold: Biology 14. PJM Aerts R. Addington RN.1097/00010694-200003000-00005 Hérault B.10.1034/j. Paine CET. doi:10. Schurgers G. Journal of IC. Duru M. 383–396. JTA Verhoeven) pp.12.1111/j. 3–16.1071/FP03176 Hacke UG.1038/ngeo905 size. Ozier-Lafontaine H (2000) On the banks and seed dispersal: important topics in restoration ecology. Menon S. 365).2307/2261481 communities. doi:10. Paula Tecco. Bond WJ (2003) Growing tall vs growing wide: tree architecture and allometry of Acacia karroo in forest.1890/03-4022 (2010) Terrestrial biogeochemical feedbacks in the climate system. Ecology 87.1999. Bonal D. Mitchell RJ. 1192–1201. Feichter seasonal water deficit. Bonal D. Chapin S III (1999) The mineral nitrition of wild plants revisited: a doi:10. The authors thank Diana Abal Solis and Juan Pablo Bellini for monitoring. Fernando Casanoves and Bryan Finegan for valuable between species within grassland communities simplifies their discussions. Thompson K (1996) Seed Adiku SGK. Poorter et al. Braddock RD. Functional Ecology 24.2008. 555–558. Jack SB. 535–545. Journal of Experimental Botany 48. Poorter L.1600-0706. In ‘Plant form and vegetation structure’.1600-0706. Parton WJ. Knight CA. Ackerly DD.x Aerts R (1996) Nutrient resorption from senescing leaves of perennials are Baraloto C. Van der Aart PJM (1991) The relation between above. Hurry VM. Chave J (2010b) there general patterns? Journal of Ecology 84.1111/j. 226–236. UK) Sheffield. Read J (1999) Methods of assessing leaf-fracture around the world for beginning to give away the secrets of their functioning properties.002 assistance in the laboratory. and leaf turgor in Acer grandidentatum populations This contribution was funded. UK) of trait measurement. Advances in Ecological Research Barkman J (1988) New systems of plant growth forms and phenological plant 30. unravelling the variable response of plant respiration to temperature. Patiño S. Kulmala M. Del Grosso SJ. 3–14. Oikos 121. Barton K. 393–405.1093/jxb/48. Cell & Environment 29. C-4 and CAM-type plants. Vesala T doi:10.x Archibald S. 2636–2660.1016/S0169-5347(00)89156-9 doi:10. by the Inter-American Institute for along a soil moisture gradient Oecologia 105. tolerance in plants.x Bakker JP. The Netherlands) Oecologia 87.x Acknowledgements Alder NN. Sack L (2012) The determinants of leaf turgor loss Agrawal AA. which were doi:10.1007/BF00328731 supported by US National Science Foundation Grants GEO-0452325 and Alder NN.1016/j. Baraloto C. Nuismer S (1997) Use of centrifugal force in the GEO-1138881 The work was also funded by FONCyT (PICT 20441 and study of xylem cavitation.19672. doi:10. 597–608. Weiss SB. Plant Science 166. Thuiller W (2012) Bradstock (2007). Cruz P (2009) Convergence in plant traits Bond. Theau J-P. doi:10. Evolution 10.2003. SECYT-Universidad doi:10. in part. Kiss AS. Oren R (2006) Adjustments in hydraulic Austin AT.1365-3040.2 meta-analysis. Chave J (2010a) Decoupled leaf and Aerts R (1995) The advantages of being evergreen. The authors appreciate helpful comments of Anten NPR. Hans Lambers and one anonymous reviewer. 551–559.00506. Many users of the previous handbook Naturalist 175. 116–126. Boot RGA. Galbacs Z (1998) New methods for determination of C-3. 1338–1347. 461–490.1016/j.2011.1111/j. Harmon ME. Sanson G. Van der Aart.CO. Functional trait variation and sampling strategies in species-rich plant doi:10. Poschlod P.024 Adair EC. Vesk and Westoby (2004). 1020–1029.2003. O’Donnell D.1365-2435. Patiño S.x 413–418. and Mark Westoby and the ARC–NZ Network of Vegetation Anderson JM. Tsigaridis K. Pellet G. Global Change Atkin OK. Bruhn D.01727.665 Nacional de Córdoba – Argentina. authors particularly thank Phil Grime and the UCPE–University of (Blackwell: Oxford. doi:10. Sperry JS. Ecology Letters 15. Oecologia 130. 402–407. Plant. Silver WL. savanna. doi:10.2010. Functional Plant Biology 32. and ecosystem-level impacts. Yoccoz NG. Harrison SP.1007/s004420100805 doi:10. doi:10.1086/649581 gave useful feedback on trait measurements. (2010). specific leaf area and microhabitat distribution of chaparral woody Ashraf M. Paine CET. De Bello F.1016/S0065-2504(08)60016-1 types. Strykstra RJ. 208–216.1016/0022-1694(79)90057-X of long-term litter decomposition in diverse climates. J. The authors also thank Marcelo Cabido. assistance with figures and diagrams and Valeria Falczuk for technical doi:10. Hart SC (2008) Simple three-pool model accurately describes patterns Hydrology 42. Ingram JSI (1993) ‘Tropical soil biology and fertility: a Function–Macquarie University.x A multi-trait approach reveals the structure and the relative importance of Belea A. (CAB International: Wallingford. and arid environments.3.2009. 250–260.12181. Donovan LA. HJ During. Hérault B. Pérez-Harguindeguy et al. Harris PJC (2004) Potential biochemical indicators of salinity plants: contrasting patterns in species level and community level analyses. Albert CH.01600. stomatal conductance. Pecot SD. Burke Aston AR (1979) Rainfall interception by eight small trees. 1–67. 449–457. doi:10. doi:10. for inspiring discussion and development handbook of methods. 525–532. Trends in Ecology & stem economics in rain forest trees.2010.1890/0012-9658(2006)87[132:PDS]2. Fishbein M (2006) Plant defense syndromes.1111/j. Lavorel S (2010) doi:10.1461-0248. On the importance of intraspecific variability for the quantification of functional diversity. and the US NSF (DEB-0620652).2005. References Oikos 102.vs. Rose CW. which greatly mechanical constraints in the economy of tree architecture. Boulangeat I. intra. Soil Science 165.01517. Acta simulation of root water extraction: examination of a minimum energy Botanica Neerlandica 45. William Ansquer P. Bartlein PJ. Beauchene J. Global Change Research (IAI) CRN 2015 and SGP-CRA2015. Pockman WT. interspecific variability in plant traits. doi:10.1038/nature05038 doi:10.1111/j.01751. Ecology Letters 13.01430. Ecological Indicators 9. Thuiller W. point and prediction of drought tolerance of species and biomes: a global S132–S149. doi:10.220 Australian Journal of Botany N.x Ackerly DD (2004) Functional strategies of chaparral shrubs in relation to Arneth A.ecolind. 369–383. 293–301. Hall SA.0. Douzet R. Sorvari S. doi:10.2012. Lavorel S.

1071/FP10010 Bhaskar R. Keitel C.123. Farquhar GD.1365-3040. Stewart E. Hill RS (2000) Increases in water potential gradient reduce community ecology and phylogenetic biology. Kono A. Biological Sciences 276. Plant Physiology 144. Smith WK.1093/treephys/26.2. Sack L (2010) Viewing leaf structure and evolution nutrient turnover. U Lüttge) Ecology and Evolution 2. Bruun HH.3732/ajb. Jerez A. McKenna J. Evidence from a low-pressure 693–715. doi:10.x Cavender-Bares J. doi:10. Trujillo PJ.x Cavender-Bares J.01314. (Eds A Läuchli. 59–67. C3 plants? Review and synthesis of current hypotheses. doi:10. Enquist BJ (2012) The leaf area shrinkage effect can bias Bucci SJ. Poschlod P (2006) Why are small seeds dispersed through animal Ochoa A.1098/rspb. (Ed. package for the integrated analysis of functional diversity. Functional Plant doi:10. Ecology Letters 12. in Environmental Sustainability 2. 555–562. Jordan GJ (2007) Leaf maximum photosynthetic rate woody plant communities.’ (Edward water droplets. Box EO (1996) Plant functional types and climate at the global scale.0028-646X.1919 Narwani A.x Volpe TJ. Dawson TE. Laskowski R (2005) Litter decomposition: a guide to carbon and Brodribb TJ. Feild TS. Burrows GE (2002) Epicormic strand structure in Angophora.1021 Barbour MM.2307/3236274 Caldwell MM. 175–183.x Brewer CA. In ‘Plant physiogical Boyce CK. 1890–1898. doi:10. Tcherkez G. Arnold: London) doi:10. doi:10.1023/A:1014905104017 Cain SA (1950) Life-forms and phytoclimate. Koutstaal B (2000) Sample preparation and scanning Cadisch G. Andrade A. Brewer CA.2000.2010. Kitajima K.tb00965. Holbrook NM. pp. Cornelissen JHC (2000) Leaf structure and moisture gradient in western Patagonia. Plassmann R. Venail P. Reich PB. Sparrow AD (2000) Resprouting as a life history strategy in Brodribb TJ. Williams DG. Pace L. Nature 486. Kembel SW (2009) The merging of Brodribb TJ. Blonder B. Ecological studies 33. 409–416. Oikos 89. Grace JB.’ (CAB International: London) Soil 218. Kozak KH. Gutiérrez MV (2003) Relations Griffiths HG. xylem conductivity in whole plants. 449–455.x conductivity method. Methods in In ‘Salinity: environment–plants–molecules’. 145–157.3. Scholz FG. Ecology Ecological Monographs 74. RW Pearcy) vein evolution was physiologically and environmentally transformative. New Phytologist 153.145 Bond WJ. Lipson R.’ (CRC Press: Boca Physiology 24.1200062 relations of tropical savanna trees: from leaf to whole plant. Pla L. Mace GM. doi:10. and venation are linked by hydraulics. Naeem S (2012) (2012) Fire-induced tree mortality in a neotropical forest: the roles of bark Biodiversity loss and its impact on humanity.1111/j. Bazzaz FA (2004) Multiple trait associations Brodribb TJ. Srivastava DS. Virginia RA (1989) Root systems.1017/S0266467407004166 persistence niche. 402–411. Cox M. Giller KE (1997) ‘Driven by Nature. Zwieniecki MA (2009) Angiosperm leaf ecology: field methods and instrumentation’.00718. Tree Blum A (1988) ‘Plant breeding for stress environments. Hall R.1365-2486. hypothesis in Psychotria marginata and Faramea occidentalis doi:10. Tilman D. Díaz S (2011) FDiversity: a software Breckle SW (2002) Salinity.0030-1299. 443–450. Bustamante M paleoclimate and ecology research. Curtis C. Lipper L. Babin D.02533.New handbook for measurement of plant traits Australian Journal of Botany 221 Bellingham PJ. Global Change Biology doi:10.’ (Chapman and Hall: recovery.1461-0248. Oikos 73. Jones N.1111/j.2307/3545728 (Rubiaceae).1046/j. Hauschild A. 1–32. 367–398 (Chapman and Hall: London) Proceedings. Bolker B.1016/S0065-2504(05)38001-9 doi:10. Current Opinion Chaney M.1007/PL00008873 between leaf trichomes. sapwood area and Böhm W (1979) ‘Methods of studying root systems.890224. Wardle DA. Ackerly DD (2006) Ecological relevance of minimum seasonal Brown S (1997) ‘Estimating biomass and biomass change of tropical forests. doi:10. Franco AC. Journal doi:10. Brando PM. 79–85. Ruiz Matthai JD.8. (2004) Functional convergence in hydraulic architecture and water 1756–1763. wood density and fire behavior. Campbell B. photosynthetic capacity during early angiosperm diversification. Santiago LS. 1021–1028. Kinzig AP. 111–131.1399-3054.2006.1111/j. doi:10. Goldstein G.2009. Mainka S. Cardinale BJ.1086/513468 range of woody plant species and types. pp. Plant and decomposition. 1771–1776.1071/FP08216 .1991. Knohl A.1007/BF02879783 of Vegetation Science 7. Oecologia 124. Functional Plant Biology 37. 34–42.x Brussaard L. tree size. Putz FE Loreau M. Klucas A. Roberts DW (2006) How should leaf area. Sloat L.’ FAO Forestry Paper 134 (FAO: Rome) doi:10. Cueto M. Gonzalez A. doi:10. water potentials. Midgley JJ (2001) Ecology of sprouting in woody plants: the doi:10. Hooper DU.1104/pp.00082. Plant. Boyle B. Feild TS. A primer.1016/j. Cell & Environment 26. Perrings C. Fine PVA.2006. Barnes C. 309–320. Cornwell WK. FL) Buckley TN. 955–962. Plant Physiology 123. Contreras K.1038/nature11148 18.1016/S0169-5347(00)02033-4 and Lophostemon (Myrtaceae) – implications for fire resistance and Bond WJ. doi:10. security: scientific challenges for a new agriculture. Furst L. 630–641.14114. traits. Nielsen KL.2003.2009.1046/j.891 Raton. Sterner M. Meinzer FC. 353–359. International Journal anatomy as related to leaf mass per area variation in seedlings of a wide of Plant Sciences 168. Cartagena P. 185–196. Simova I. leaf turgor and xylem vulnerability in eight photosynthetic tissues generally 13C enriched compared with leaves in tropical dry forest trees. Brodribb TJ. 233–237.2041-210X. 1–17. 488–498.00975. 45–51. Gellagos J. Thornton J. Advances in Ecological Research 38. doi:10.2011. Ellsworth DS.1111/j. Richter R. Barber B. Brodribb TJ. Fisher M. 53–77 (Kluwer Academic Publishers: London) doi:10. 476–486.03. Argentina.2008. Daily GC. Vogelmann TC (1991) Functional interaction doi:10. Bond WJ.x Casanoves F.1104/pp. Venus JC (1981) ‘The biometry of plant growth. Aguilar. Mosier N. Botanical Review 16. Rabbinge R. Physiologia Plantarum 127.’ stomatal conductance vary with tree height to maximize growth? Tree (Springer: Berlin) Physiology 26. doi:10.1890/03-4007 Letters 13. Puyravaud JP. leaf wettability and the optical properties of Causton DR. Caron P. doi:10.1365-3040. doi:10. Plant. Balch JK. Duffy JE.1034/j. London) doi:10.x Biology 36. Cell & Environment 14.1111/j.101352 Berg B. Feild TS (2010) Leaf hydraulic evolution led a surge in leaf in relation to habitat differentiation among 17 floridian oak species. 199–213. guts: large numbers or seed size per se? Oikos 113. Wright IJ (2009) Why are non- between stomatal closure. Journal of Tropical Ecology 23. doi:10. Van Wilgen BW (1996) ‘Fire and plants.1461-0248.007 Lamb M. Russakoff B. Coe M. Plant litter quality and protocol for computerised analysis of root length and diameter. Nepstad DC. Midgley JJ (1995) Kill thy neighbour: an individualistic Burd M (2007) Adaptive function of drip tips: a test of the epiphyll argument for the evolution of flammability.1600-0706. Pulleman M (2010) Reconciling biodiversity conservation and food Beaucage B. Thompson TT.x doi:10.2010.1093/treephys/24.x Bouma TJ. 891–899. Stagg R. Larigauderie A. Núñez CI (2007) Patterns of leaf wettability along extreme Castro-Díez P. from a hydraulic perspective. Navabi M. 635–662.cosust. doi:10.2001. Bushey D.1111/j. halophytes and salt affected natural ecosystems.1111/j.00299. Eucalyptus doi:10. doi:10. Trends in Ecology & Evolution 16. Edwards EJ. McIntyre C. Di Rienzo JA. Feild TS. Aubyn HS.01410. Buzzard V. Christman MC. Cernusak LA. American Journal of Botany 99.107.

Riéra B.001313 Oecologia 118.1992. allocation response to environmental stress. doi:10. Kira T. 619–627. Jónsdóttir Grime JP.01916. Schmidt IK. Michelsen A.1111/j. Pérez-Harguindeguy N.2009. Wardle DA. Van Bodegom PM. structure and function of Hawaiian Metrosideros polymorpha at high vs. Cambridge Philosophical Society 78. Coley PD (1987) Patrones en las defensas de las plantas: ¿por qué los Vaieretti MV. van der Heijden MGA. Lescure J-P. doi:10.001305 Ecology 84. Bishop ID (2010) Towards better prediction of doi:10.x pH as a new plant trait: can it explain variation in foliar chemistry and Chen N. Onipchenko VG. doi:10. DeWilde L. Calef M. New Phytologist 175. Hik DS. Eviner VT. Vendramini F. Bhaskar R. allocation and leaf traits in Consequences of changing biotic diversity. ontogeny. Oecologia 74. Holbrook NM (2007) Diversity of hydraulic traits in doi:10.1038/35012241 111. Morgan HD.1469-8137. 205–209. Ginnett TF (1998) Spines protect plants against browsing by Biology 15. Annual Review of Cornelissen JHC (1999) A triangular relationship between leaf size and seed Ecology and Systematics 11. 2431–2449.1007/BF00380050 wood fates across the globe: rotted. Fölster H. doi:10. Naylor RL (2008) Increasing wildfire in Alaska’s control litter decomposition rate across species and life forms in regional boreal forest: pathways to potential solutions of a wicked problem. Tognetti R.110182.1146/annurev. Tyree MT (1992) Use of positive pressures to establish Quested HM. seedlings of a wide range of woody plant species and types. Andalo C. Lunch CK (2007) Adjustment of implications. rates in a wide range of temperate plant species and types. Puig H. Sack L. Pérez-Harguindeguy et al.x nine Cordia species growing in tropical forests with contrasting Cornelissen JHC. Cairns MA.1111/j.1007/s004420050371 seed dispersal by animals: conceptual frameworks and process-based Cooper DJ. Lovecraft AL. Kurokawa H.2007. Hobbie SE. Hartley AE. Oecologia doi:10. 219–221. (2011) Leaf pH as a plant trait: species-driven rather than soil-driven doi:10.1111/j.1046/j. 19–102.2307/3220956 Godoy O. doi:10.01323. Wada N. Santiago LS.1007/s00267-002-2808-2 Press: Princeton. Van Logtestijn RSP.x Chave J. or consumed? Global Change Cooper SM. doi:10.1111/j. Biological Reviews of the Laundre J. 21–28. Hobbie SE. Australian Journal doi:10. Cerabolini B (1999) Leaf structure and defence Rupp TS.1111/j. response patterns of Populus deltoides to alluvial groundwater pumping. Aerts R.2010. Pereira B. doi:10. Diaz S (2000) AP (1997) Foliar nutrients in relation to growth.x Coley PD (1988) Effects of plant growth rate and leaf lifetime on the amount Cornwell WK. Dorrepaal E. Naylor RL. Cordell S. carbon cycling processes among Subarctic plant species and types? Cell & Environment 15. Lucas PW. S78–S92. Chambers JQ. Gerdol R. Zaveleta ES. size among woody species: allometry. Annual Review of Cornelissen JHC (1996) An experimental comparison of leaf decomposition Ecology and Systematics 13. Pérez-Harguindeguy N. Cornwell WK. Totland Ø. Díaz S. Chabot BF.1111/j.1469-8137. Swenson NG.x variation. doi:10. Choat B. doi:10. Craine JM (2009) ‘Resource strategies of wild plants. Callaghan TV. Pugnaire F (1993) Evolution of suites of traits in Cornelissen JHC. Cornelissen JHC. Castro-Díez P.tb00992. Lavorel S. Ong JSY.1469-8137. Westoby M (2008) Plant species traits are the predominant herbívoros prefieren ciertas especies? Revista de Biologia Tropical 35. 233–260. 1163–1170. Jansen S. Vitousek PM. Gurvich DE.1461-0248.1111/j. Natcher Cornelissen JHC. Allison SD.1007/s00442-005-0269-z doi:10. Werger MJA. Quested H. Thompson K precipitation. Huntington H. Soudzilovskaia NA. Klein JA.1365-2435. Zhao X. Quested HM. Díaz S.1017/S1464793102006000 Soudzilovskaia NA. Letters 11. Pausas JG. New Phytologist 121.13. Van Bodegom P.2307/2261479 Chapin FS III (1980) The mineral nutrition of wild plants.2010.100. Rowland Reynolds HL. 573–582.x small climbing mammals. Sala OE.x low precipitation. Eggleton P. Wirth C.2008.x Environmental Management 31. McGuire D. 315–326. 248–255.222 Australian Journal of Botany N. Bauhus J. Weedon JT. Sandvik SM.01219.1086/285524 Journal of Ecology 84.1461-0248. Hill J. Turner IM (1992) doi:10. Hooper DU.1104/pp. 87–99.x Leaf fracture toughness and sclerophylly: their correlations and ecological Cornwell WK. 215–226. Scott ML (2003) Physiological and morphological models. Zavaleta E. Read J. Marzano B. Gudmundsson J.00430. Ecological Research 6. Hunt R (1996) Seedling growth.110180. burned. Klein Alatalo J. Ecology Letters 10. Díaz S.01765. Nature 405. Kazakou E.1365-2435. Karlsson S.x Cornelissen JHC. Journal of doi:10.1641/B580609 doi:10. 531–536. Chiba Y (1991) Plant form based on the pipe model theory II. Chatain A. Gwynn-Jones Identification. Ecology 151–164. Stuart Chapin F. MOL Team (2007) Global negative vegetation analysis of ramification in morphology.1999. measurement of plant functional traits worldwide. Eamus D. 1065–1071. Lavorel S. Aerts R. 531–540. Lewis SL.’ (Princeton University doi:10. Fresco N.2307/2261337 Chapin FS III. Nelson JL.205 Gurvich DE. Callaghan TV. DC. control on litter decomposition rates within biomes worldwide. Trainor SF. Hoorens B. Sibma F. 686–698. doi:10.1146/annurev. 191–200. 597–610.1071/BT02124 Chave J. floras on two continents.1.1365-2435. Van Logtestijn Cherubini P.x Transactions of the American Microscopical Society 26. Castro-Díez P. Sack L. Broekman R. 119–148. Towards a worldwide wood economics spectrum.01051. Ogawa H.01747.1007/s00442-005-0100-x of Botany 51. doi:10. Clements ES (1905) The relation of leaf structure to physical factors. measurement and interpretation of tree rings in woody D. American Naturalist 142(Suppl). Oecologia 145. Schoch W. Callaghan TV. Scarff F. Buchmann N.1992. Magnusson B. CM. French K.1461-0248. 234–242. Bioscience 58. Díaz S. New Phytologist 143. Preston and type of anti-herbivore defense. Van Logtestijn RSP. feedback to climate warming responses of leaf litter decomposition rates doi:10. 1063–1071. 460–469. ecology and taxonomy. Shaver GR. vulnerability curves. Cornelissen JHC. Functional Ecology 25. Functional Ecology 21. Yamakura T (2005) Tree allometry and improved estimation of Poorter H (2003) A handbook of protocols for standarised and easy carbon stocks and balance in tropical forests. Higuchi N. Coomes DA.1365-2486. Cousens RD.02137. 335–380. Mack MC.1111/j. Huntington O. Quantitative Welker JM.2007. Solheim B. Tan HTW. Ray L. Nelson BW. Cornelissen JHC. Allison SD. 755–765. Cochard H. and leaf attributes in a wide range of woody plant species and types. doi:10.1007/BF02353867 in cold biomes. Brown S. Gartner BL.1007/s004420050725 Chapin FS III. Cornelissen JHC. Black TA (1992) Defining leaf area index for non-flat leaves. Reich PB.tb01131. Oecologia 113. doi:10. Molau U. species from mediterranean climates. doi:10. Garnier E. Aerts R (2006) Foliar 351–366. 449–455. Wright IJ.01285. NJ) . doi:10. 229–259.1111/j. Hofgaard A. Functional Ecology 24. Choong MF. Amatangelo K. Innes JL (2003) RSP. Gwynn-Jones D. 421–429. Cabido M. Autumn K. Ecology Letters 12. Zanne AE (2009) Plant traits and doi:10. doi:10.2009. Fromard F. Plant Physiology 100. Pérez-Harguindeguy N. Tolvanen A.1365-3040. Zanne AE (2009) Cornelissen JHC. Cruizat P. Reich PB. Garnier E. Plant. doi:10.11. ter Steege H. Press MC. Eviner VT. Hicks DJ (1982) The ecology of leaf life span.1007/s004420050259 Chapin FS III. Bräker OU. D’Amico DR.1111/j. Oecologia 147. Stenström A. Brovkin V. doi:10. Van Rheenen JWA.

Journal Ellsworth DS. Bogaard A. Guerrero-Campo J.x Davis SD. Band SR. Funes G. Cerling TE. UK) of future climate and atmospheric CO2 content for regional Cramer W. 23–27.1016/S0065-2504(08)60005-7 doi:10. Oecologia 112. Adams GC Eamus D. Global Change Biology 16. Kazemi-Saeed F.1998. De Bello F. 954–962. Hodgson JG. Puigdefábregas J (1998) results from six dynamic global vegetation models. doi:10. Agricultural and Forest Meteorology 91.1016/j. Zak MR (2004) The ETH. Jalili A.’ (Backhuys Publishers: Leiden.1046/j.x terrestrial ecosystem structure and function to CO2 and climate change: Domingo F. Reier Ü.930210. 187–200 (Academic Press: London) De Kroon H. Stiftung Rübel 37. 271–288 (Cambridge University Press: Cambridge.02. Tree Physiology 22.2001. architecture and the spatial and temporal variability of nitrate supply. Measurement Science & Dunbabin V. Castro-Díez P. Nielsen SL (1996) Broad-scale future climate.1007/BF02858075 Elberse WTH. Bondeau A. Grime JP. doi:10. Crocker MC. Valente F. Knops J (1999) Measurement Diemer M (1998) Life span and dynamics of leaves of herbaceous perennials of leaf longevity of 14 species of grasses and forbs using a novel in high-elevation environments: ‘news from the elephant’s leg’. Grigulis K. 107. 357–373. Harvey V. (Eds DC Coleman. Thuiller W. Edwards EJ.1016/S0065-2504(01)32012-3 failure. Functional Ecology 12. 204–211. Thornton PK (2010) Implications pp. Gunn TC. Moro MJ. . Kerenga K.1469-8137.00207. Gerhold P.1093/treephys/22.1111/j.x doi:10.04. 274–285. Betts RA. Falczuk V. Duarte CM.tree. J De Vogel EF (1980) ‘Seedlings of dicotyledons. Proceedings of the National Academy of Sciences. plant traits that drive ecosystems: evidence from three continents. In ‘Encyclopedia of hydrological sciences’. 295–304. 1–157. (Eds TM Smith. Pérez-Rontomé MC.0269-8463.2004.1 Incorporating plant functional diversity effects in ecosystem service Enquist BJ (2002) Universal scaling in tree and vascular plant allometry: assessments. Annals of Botany 106. Jones G.1093/treephys/10. saccharum seedlings in contrasting natural light and water regimes. Ellenberg H.01997.2001.2007.1111/j. De Bello F. Ekologia 4. doi:10.1365-2486. Ramankutty Africa. In ‘Carbon isotopes techniques’. Diefendorf AF. biogeography and ecosystem services across East PM. 285–299. Basconcelo S. Lavorel S. Maestro-Martínez M.1046/j. Sand-Jensen K. Smith B.x canopies.1071/WF01003 ‘Plant functional types’. International Journal of Wildland Fire Cramer W (1997) Using plant functional types in a global vegetation model.1365-2486. Reich PB (1992) Water relations and gas exchange of Acer of Vegetation Science 15. Rengel Z.1073/pnas.00071.3732/ajb. Biological Conservation 143. Grigulis K. 475–481. UK) doi:10. Prentice IC. Griffiths H. Panov PI (2001) Pyric properties of some dominant doi:10. 9–17. HH.1999. The Netherlands) Publishers: Dordrecht.1365-2435. Shugart. 627–634 (John Wiley and Sons: Chichester. Global Change Measurement and modelling of rainfall interception by three semi-arid Biology 7. Sitch S.x pp.00411. Methods in Ecology and Evolution 2. PW Rundel) pp. Berin DM. Boustani S. nutrient availabilities. and climate.New handbook for measurement of plant traits Australian Journal of Botany 223 Craine JM. Advances in Ecological Research 32. Earnshaw MJ.1073/pnas. Chapin S III (2001) The relationships Dilcher DL (1974) Approaches to the identification of angiosperm leaf among root and leaf traits of 76 grassland species and relative abundance remains. Kowsary E.0704716104 cells to ecosystems.1600-0706. doi:10.002 S.1007/s004420050311 Del Tredici P (2001) Sprouting in temperate trees: a morphological and Eissenstat DM. Yanai RD (1997) The ecology of root lifespan.2307/2389891 Pérez-Harguindeguy N. Friend AD. and morphology of eight grass species from habitats with different Montserrat-Martí G. Berendse F (1993) A comparative study of the growth Díaz S. 20684–20689. 281–300 (Kluwer Academic Publishing and Documentation: Wageningen. approach. doi:10. Abbas-Azimi R. doi:10. Lucas PW (1996) A portable fracture doi:10. 280–288. Tilman D. Gash JHC (2005) Evaporation of intercepted rainfall. 289–295.1111/j.1088/0957-0233/7/6/016 root systems: efficiency of nitrate uptake is dependent on root system David JS. Lavorel S. In ‘Plant physiological clonal plants.1034/j. Robson MT (2007) Physiology 10. 197–206. In 10. 163–174. Sánchez G. Lee PKD. Smith B. (Eds RW Pearcy. 5738–5743. N. Shirvany FA. Oecologia 84. Villar-Salvador P.00827. Lewis SL. American Journal of Botany 89. chilling tolerance De Bello F. Van Groenendael JM (1997) ‘The ecology and evolution of Ehleringer JR.15-16. Functional Ecology 7.2041-210X. (Ed. Freeman KH (2010) Global doi:10. conditions.1016/j. Kucharik C. with revised subdivisions. Portwood KA. USA toward a general quantitative theory linking plant form and function from 104.1. Asri Y. Yuen TDB.x Mediterranean vegetation species. New Phytologist 142. The Netherlands) De Witte LC. 617–640.1093/aob/mcq191 doi:10. Müller-Dombois D (1967) A key to Raunkiaer plant life forms Hynd A. Prior L (2001) Ecophysiology of trees of seasonally dry tropics: (2002) Shoot dieback during prolonged drought in Ceanothus comparisons among phenologies. Osmond CB (2000) Stable isotopes. 275–292. (Rhamnaceae) chaparal of California: a possible case of hydraulic 113–197. Brovkin V. Dolezal J. Oikos 93. Foley JA.biocon.2009. doi:10. Dimitrakopoulos AP.00383. Janecek doi:10. 1–60. Cox biogeochemistry. HA Mooney. Berichte des geobotanischen Institutes der Romo-Díez A. Trends in Ecology & Evolution 22. Donoghue MJ (2007) The relevance of phylogeny to doi:10. 820–828. Lêps J (2011) Quantifying the relevance of intraspecific trait variability Ehleringer JR (1991) 13C/12C fractionation and its utility in terrestrial plant for functional diversity. doi:10. The Netherlands) ecology: field methods and instrumentation’. Fisher V. Cornelissen JHC.1045 patterns in leaf 13C discrimination and implications for studies of past and Enríquez S. MG Anderson) Functional Ecology 18. Hamzehee B. de Torres-Espuny L. studies. Cabido M. Wing SL. Charles M. Helliker BR (1997) C4 photosynthesis.022 studies of global change. Ewers FW. Yazdani S. Lomas MR. Botanical Review 40.1016/S0168-1923(98)00068-9 toughness tester for biological materials. Tilman DG.x Craine JM.5.89. Pärtel M (2010) A biodiversity and cell sap osmotic potential values of grasses along an altitudinal monitoring framework for practical conservation of grasslands and gradient in Papua New Guinea. Advances in ecological review Botanical Review 67. 859–870. pp. 1–20. Dehghan M. Still CJ. Carver KA. Lavorel S. Stöcklin J (2010) Longevity of clonal plants: why it matters Ehleringer JR. Reich PB.2009. Zarrinkamar F. FI Woodward) Doherty RM. Sperry JS. Vendramini Eliáš P (1985) Leaf indices of woodland herbs as indicators of habitat F. B Fry) doi:10. Khoshnevi M.1046/j. Shaw S. 413–425. doi:10. 223–229. Sitch S. Siavash B. Albert CH. 243–249. doi:10. Mueller KE.’ (Centre for Agricultural Ehleringer. 1045–1064. Woodward FI. shrublands. doi:10. Proceedings of the National Academy of Sciences. White A. Quétier F.1007/BF02860067 along fertility and disturbance gradients. 56–73. Brenner AJ. Diggle AJ (2004) Simulating form and function of Technology 7. Tree Díaz S. Wedin DA. doi:10. Ecological Research 27.820 Broadmeadow MSJ (1990) Photosynthetic pathway. doi:10. 121–140. Thompson K. and how to measure it. Young-Molling C (2001) Global response of doi:10. atmospheric CO2.0910513107 Oecologia 108. Darvell BW.2010. USA comparison of photosynthetic rates across phototrophic organisms. Koch PL.

Basconcelo S. New Phytologist 152. doi:10. 440–446. stem and root tissue strategies Givnish TJ (1987) Comparative studies of leaf form: assessing the relative across 758 Neotropical tree species. Díaz S. In ‘Seed fate: Gaucherand S.00239.1086/415032 (1980) Age states of plants of various growth forms: a review. In ‘On the economy and ecosystem functioning in grasslands: a standardized methodology of plant form and function’. S Jain. Plant functional markers capture ecosystem properties during secondary Ecological Applications 8. methods and applications for paper 57 (FAO: Rome) agroecology. Methods and interpretation of Garnier E.2012. Shipley B.1093/aob/mcl215 Fitter A (1996) Characteristics and functions of root systems. (Eds GH 109–114.1469-8137.00563. 113–185. Farquhar GD. Annales Botanici Fennici 36.1111/j.x Kigel J. Billès G. Etlinger MG. communities Fisher JB (1986) Branching patterns and angles in trees. 313–337. Navas 177. Aerts R (2010) Evidence performance of landscape plants. Golodets C. Doležal J.tb04687. The Quarterly Gatsuk LE. Papanastasis VP. Quétier F. (Ed. doi:10. 1206–1221.0269-8463. Van Breemen N. Roumet C. Rusch dispersal of plants and animals on sheep in calcareous grasslands. R Dirzo. Dolezal J.1046/j.1987.x pp. doi:10.04. 665–675. Joffre J. Laurent G (2001a) A standardized protocol and Plant Molecular Biology 40. New Phytologist Garnier E. In ‘The halophytes. Laurent G. 242–243. Lavorel S (2007) New method for rapid assessment of predation. Gill AM. Lepš J.2001.1111/j. Skarpe C. Lavorel S. Neill C.x Wallingford. doi:10.1038/334242a0 Jouany C. Aerts R.28. Vorontzova LI. Grigulis K. 598–611. Roumet C. Lehsten V. Functional Ecology 26. Hubick KT (1989) Carbon isotope doi:10. 2630–2637. Brokaw NVL (1996) Forest damage and recovery from doi:10. doi:10. 945–963. Zaugolnova LB. 25–55 (Cambridge University Press: Cambridge. Clipson NJW (1986) Halophytes. New and potential consequences for tree biomechanics.x catastrophic wind. Phytologist 179.1111/j. Golodets C. Ansquer P.cosust.1 pp. Colmer TD (2008) Salinity tolerance in halophytes.1890/03-0799 Fischer M. 727–737. doi:10. Berthelier P. Funes G. Toussaint J-P (2004) within temperate forests: species effects on soil carbon and nitrogen. Freitas H. Annual Review of Plant Physiology 28. Ecology 85. Theau J-P.1046/j. Balun L (2008) Xylem hydraulic and photosynthetic function of doi:10. (Eds P Forget. Ansquer P. Annals of Botany 99. University Press: Cambridge. 88–94. Baraloto C (2012) Leaf. Agronomy for Sustainable Development 32. Yeo AR (1977) The mechanism of salt tolerance in Garwood NC (1996) Functional morphology of tropical tree seedlings. TJ Givnish) pp. Zarovali MP (2007) Journal of Applied Ecology 33. Orians. Everham EM. Theau J-P.1071/WF04039 98.1442-9993. competitive ability from plant traits. Troke PF.2007. JH Cushman) pp. PH Raven) across Europe.1007/s13593-011-0036-y discrimination and photosynthesis. 66–74. Tree Physiology 24. 101–126 (Springer: Berlin) Gardner RO (1975) Overview of botanical clearing technique. Evosystem services: an evolutionary perspective on the links between Garnier E. doi:10. A Eshel. Current Opinion in Environmental in congeneric annual and perennial grass species. doi:10.x FAO (1999) ‘Soil salinity assessment. Annual Review of Plant Physiology Garnier E.1890/08-0418. 927–936. 99–105. Ecology 90. MD Swaine) pp.1365-2435. Robson M.96082. Papadimitriou Fischer S. New Phytologist doi:10. Ehleringer JR. Bigelow SW (1996) Plant life forms and ecosystem functioning.1007/BF02857920 of Lachemilla pinnata (Rosaceae) in different plant communities of Ewel JJ. 2nd edn.1046/j.1523-1739. 493–523 (Cambridge and lessons from an application to 11 European sites. In ‘Plant roots: Gartner BL (1995) ‘Plant stems: physiology and functional morphology. Feild TS. doi:10. Zylstra P (2005) Flammability of Australian forests. Grigulis K. Bellmann A. Fine PVA. 362–373.tb04036. plants.x Evans GC (1972) ‘The quantitative analysis of plant growth. succession. Lepš J.x Gnetum (Gnetales) species from Papua New Guinea. Freitas H. Wenny D (2005) How to elucidate seed fate? A review of methods Ecology 68. Stain Faith DP. 1–20 (Academic Press: San Diego. doi:10.02531.1111/j. Garnier E. Pérez-Harguindeguy et al. Debussche M. Castro H.01781. Stöcklin J (1997) Local extinctions of plants in remnants of Garnier E. 69–83. Conti E. Beall FC (2004) Development of a laboratory protocol for fire Freschet GT. Cornelissen JHC. Cabido M (1999) Seed bank dynamics doi:10. Cruz P. mountain grassland in central Argentina. Mooney HA (1986) The photosynthesis–nitrogen relationship in wild traits. 131–160. Keddy PA (1988) A comparative approach to predicting Fortunel C. Journal of Ecology 13. UK) 967–985. doi:10. 688–695. Quested H.1111/j. Ducout B. Australian Papadimitriou M.060189.1111/j.pp.2009. Journal of Forget P. Laurent Finzi AC. Castro H. S Vander Wal) pp.11. doi:10. Sternberg M. Lehsten V.1093/treephys/24. J the functional composition of herbaceous plant communities.224 Australian Journal of Botany N. In ‘On the economy of plant form and function’. CA) (Marcel Dekker: New York) Gartner BL. doi:10. TJ Givnish) doi:10.1016/j. Roumet C. Navas M-L. Beinlich B (1996) Experimental studies on the M. Papanastasis VP. (Eds Y Waisel. Eriksson O. Pakeman R. 11. 479–488. UK) Gaudet CL. Vile D. In ‘Topics in plant of land use changes and climate on litter decomposability of grasslands population biology’.1997. Forestry 68.2011. Debain S. Blanchard A.2307/2259429 used to study seed removal and secondary seed dispersal.02306. Cruz P. Sustainability 2.002 725–736.0028-646x. U Kafkafi) pp.’ FAO irrigation and drainage functional plant ecology: concepts.2010. Zhukova LA Review of Biology 61.01615. 379–394 (CABI Publishing: Ecology 32. doi:10. Moore JC. G.2007.40. GB Johnson. Donoghue MJ (2010) Technology 50.002443 Functional Ecology 15. doi:10. Quested H. Aubry D. for the determination of specific leaf area and leaf dry matter content.1111/j. Cortez J.060177.1146/annurev. Sternberg M. UK) Garnier E. Quétier F.x 1239–1250. P Hulme. Kleyer M.x 106.1469-8137.01913. Eriksson O. Hendry AP. extensively used calcareous grasslands 1950–85 Conservation Biology Fortunel C. ecology of tropical forest tree seedlings’. Gardiner BA (2004) Gas in stems: abundance Flowers TJ. 503–537. Canham CD (1998) Canopy tree-soil interactions G. doi:10. Laurent G (1994) Leaf anatomy. Jouany C.000513 (Parthenon: New York) Flowers TJ. Zarovali M (2009) Leaf traits capture the effects Givnish TJ (1979) On the adaptive significance of leaf form. OT Solbrig.2001.1111/j. Lavorel S. doi:10. 675–696.1365-2745. Kleyer M. Botanical Review 62. specific mass and water content biodiversity and human well-being. Smirnova OV. Van Logtestijn RSP. Functional Ecology 26. International Journal of Wildland Fire of the ‘plant economics spectrum’ in a Subarctic flora. 365–399.1365-2435. 1153–1161.2008. doi:10. Nature 334.x M-L (2001b) Consistency of species ranking based on functional leaf Field C.1994. Meier T. Robson M.x . Roumet C. Kazakou E. roles of selective pressures and phylogenetic constarints. Pakeman R. Yahara T. Thébault A. Cornelissen JHC (2012) A plant economics spectrum of Scientific Publications: Oxford. dispersal and seedling establishment’.pp. Kigel J. Navas M-L (2012) A trait-based approach to comparative electrical conductivity measurements. (Ed. New Phytologist 128.02020.’ (Blackwell Freschet GT.2307/2404699 Assessing the effects of land-use change on plant traits.’ the hidden half’. Hajibagheri MA.1146/annurev. Kazakou E.1469-8137.1469-8137. 375–407 (Macmillan: London) Fortunel C. Austral Lambert. In ‘Biodiversity and ecosystem processes in tropical forests’. 56–65. Meier T. (Ed. 59–129 doi:10. Magallón S.1239 Flowers TJ. 89–121. Poschlod P. UK) litter decomposability. (Ed.

(Ed. M Hadley) pp.1046/j. doi:10. Davis SD. Ashenden TW. de Torres Espuny L. doi:10. MJ Crawley) pp. Grime JP.1086/303201 A laboratory manual. Hattersley PW. Ecology 76. 217–223. Sharafi M. Falczuk V. Ross. Journal of Ecology and Systematics 13. Pérez-Harguindeguy N. Oikos 79. Journal of Ecology 63. Cornelissen JHC. 1801–1812.’ CC. 1337–1345. doi:10. UK) Castro-Díez P. Thompson K.x analysis.1111/j. Charles M. Reiling K. Liu Z. 239–245. 284–324 (Blackwell biogeochemistry and ecosystem function.1093/treephys/21.2000.1442-9993.2307/1941205 correlate with leaf structure among deciduous and evergreen tree species. Annual Review of and from competition among plants and among animal.1046/j. Forest Science 20. Hodgson JG (1996) Evidence of a Carter G. Booth RE. doi:10. Thorpe PC. doi:10. Miquel S (1990) Seedling types and plant establishment in an Tree Physiology 21.1078/1433-8319-00050 contrasting plant functional and chorological types. doi:10. 1341–1346. Running SW (1978) Temporal and spatial experiment with three Berberis species. Green DS. Lamont BB (2002) Relationships between physical and chemical dry forest. Tasker DE. Cerabolini BEL. The Netherlands) smong species in a herbaceous plant community. 201–228. variations in the water status of forest trees. Science: Oxford. Bossard Holbrook NM.1146/annurev. of grass species. Askew AP. Bogard A.1111/j. (Eds KS Grime JP (1979) Competition and the struggle for existence. Kruger EL (2001) Light-mediated constraints on leaf function doi:10. Naturalist 153. 219–240 (Wiley: Chichester.1365-2435. earth systems and society’. Austral Ecology 36. 32–46.01015. P Kabat) pp. Romo-Díez A. The functional approach to plant growth doi:10. (Ed. 393–422. J Dodson) pp. doi:10. Jones G. Thorpe C demography and evolution.1469-8137.00540. Oikos 85. variability: a recipe for grass-tree coexistence in savanna.2010. Bartlein PJ (2010) Fire in the Earth system. In ‘Grass (Eds JD Tenhunen. Cabido M. Grime JP. (2011) Differential effects of plant diversity on functional trait variation In ‘Plant ecology’.00612. 1139–1145. Davison AW. Hall W. In Ibarra-Manriquez G.2002.1111/j. Díaz S (2011) Device for the standard and petioles of C3 flowering plants. Pérez-Rontomé MC. Thompson K.1093/aob/mcq220 Hulme PE (1998) Post-dispersal seed predation: consequences for plant Gurvich DE. Zwieniecki MA (2005) ‘Vascular transport in plants. In ‘Reproductive ecology of tropical plants’. 614–632. 213–229. 75–84 (CRC Press: New York) Grubb PJ (1992) A positive distrust in simplicity – lessons from plant defences Howe HF. 216–222.x significance in a local flora. Jackson UB (1999) Allometry of constitutive defense: University Press: London) a model and a comparative test with tree bark and fire regime. Trolliope WSW (2000) Fire.x within and across individuals and species: an example from a Costa Rican Hanley ME. Guerrero-Campo J. Grime JP. Lipowsky A. Thornton JD (1970) Food selection by the snail Cepaea 282–294.1365-2028. New Phytologist 162. 100–113.00434. Wilson PJ. Graça MAS. 218–223. New Phytologist 180. doi:10.2307/2260852 doi:10. doi:10. Hodgson JG.’ (Springer: Dordrecht. Blythe GM. Band SR. Martínez Ramos M.New handbook for measurement of plant traits Australian Journal of Botany 225 Gowda JH. Jalili A. Adams DC. In ‘Plant ecology’.’ (Edward Arnold: London) Harrison SP.1016/0022-1694(85)90209-4 doi:10. Ewers FW (2008) Comparative community Herwitz SR (1985) Interception storage capacities of tropical rainforest physiology: nonconvergence in water relations among three semi-arid canopy trees.2307/2258728 C-S-R plant functional types: a soft approach to a hard problem. Oikos 77.2009.2008. measurement of shoot flammability in the field.1016/j. Royo Pla F causal connection between anti-herbivore defense and the decomposition (2011) Is leaf dry matter content a better predictor of soil fertility than rate of leaves. In ‘Integrating hydrology. Hodgson JG.1038/415451a 821–829.x .2002.08. Jones CG (1997) Plant chemistry and herbivory.’ (Princeton University JML. 237–252. UK) dynamics’. UK) evolution and domestication’. Díaz S.x Functional Ecology 16. attributes of congeneric seedlings: how important is seedling defence? doi:10. In ‘Animal populations in relation to their food resources’. Rorison IH. Bertone GA.1046/j.001 a0001–z0001. Watson L (1992) Diversification of photosynthesis. 38–116 (Cambridge Jackson JF.x Gowda J. Westley LC (1997) Ecology of pollination and seed dispersal. Montserrat-Martí G. doi:10.00435. is green. or why the world Jackson RB (1999) The importance of root distributions for hydrology.001221 Gubsch M. Hodkinson DJ.2004. Acta Oecologica 26. YP Kalra) plants. UK) doi:10. Journal of doi:10. Grime JP (1993) ‘Methods in comparative plant ecology. Buchmann N. 466–474. Hannah MA. 73–99 (Blackwell Scientific Publications: Oxford. doi:10. Schulze ED. Bond WJ. (Eds RM Anderson. Pratt RB. 261–282 (UNESCO/Parthenon: Carnforth. African Journal of Ecology 41. Whitehouse J Horneck DA.02222.1469-8137. Hynd A. resprouting and tortilis Hayne. (Ed.110182.x Ecology 88. American Journal (Springer: Dordrecht.1093/aob/mcr225 Hendry GAF. The Netherlands) of Botany 88. Band SR. Press: Princeton. 21–48 functional types in a lowland rain forest in México. 157–169.actao. Ecology 80. Annals of Botany 107. Marlon JR. Grime JP. Functional Ecology 24. Perspectives in Plant Ecology. Hodgson JG. doi:10. Smallwood J (1982) Ecology of seed dispersal. Hunt R. Roscher C Howe HF. Evolution (2002) Foliar resistence to simulated extreme temperature events in and Systematics 1. Nature 415. Sutton F. 585–610. Spencer RE.x Hibberd JM. 259–281. Mackey Horn HS (1971) ‘The adaptive geometry of trees. 123–139 Hodge A (2004) The plastic plant: root responses to heterogeneous supplies (Blackwell Scientific Publications: Oxford. Hladik A. NJ) Fraser W.1111/j. Gupta PL.1365-2486.’ (Chapman and Hall: London) Jacobsen AL. Cornelissen JHC. UK) of nutrients. Shipley B.2307/3546494 nemoralis L. 451–454. doi:10. Wilson P. Raffaele E (2004) Spine production is induced by fire: a natural Hinckley TM. Thompson K (1999) Allocating doi:10. BD Neal AM. (1997) Integrated screening validates primary axes of specialisation in In ‘Handbook and reference methods for plant analysis’. Hunt R. American Hendry GAF.2003.2307/3558356 Hartley SE. A Watson) pp. Grime JP. MJ Crawley) pp. Hand (Elsevier Academic Press: London) DW. Global Change Hulshof CM.2307/3545938 specific leaf area? Annals of Botany 108. Palo RT (2003) Age-related changes in defensive traits of Acacia Higgins SI. Cornelissen JHC. (Ed. Hunt R (1982) ‘Plant growth curves. Lassoie JP. Reader RJ. (Ed. (Ed. LR Taylor) pp. Cooper JEL. Matthews N.02554.01614. GP Chapman) pp. Swenson NG (2010) Variation in leaf functional trait values Biology 8. 489–494.13. Hillier SH. Werger MJA (1995) Canopy structure and shoton slux sartitioning decomposition. doi:10. shrub communities. Schmid B. 9–24. Campbell BD.2307/3546011 pp. 262–283 (Blackwell: Oxford.18. Hunt R (1975) Relative growth-rate: its range and adaptive doi:10. Palmer C. In ‘Population Bawa.2004.1046/j. Miller RO (1998) Determination of total nitrogen in plant tissue.1365-2745. Mowforth MA. Oyama K (2001) Seedling ‘Changing climates. Gessner MO (2005) ‘Methods to study litter Hirose T.1341 African rain forest.1365-2435. Journal of Hydrology 77. Quick WP (2002) Characteristics of C4 photosynthesis in stems Jaureguiberry P. UK) ecosystem dynamics and biogeochemistry in complex landscapes’. doi:10. Bärlocher F.

Freschet GT. White S. Wright SJ. Medlyn BE. 149–160 (Springer-Verlag: Berlin) Kitajima K.1104/pp. Klimešová J (2005) Clonal traits. Baldocchi D. plants: a search for physiological causes and ecological consequences. Kocacinar F. Kleyer M (2003) Life-history traits of the northwest Coomes D. Hermy M. Atkin O. doi:10. 708–721. Eriksson O. doi:10. Blanco CC. doi:10.1365-2435.1093/aob/mcp202 Larson DW (2001) The paradox of great longevity in a short-lived tree species.2009. Fyllas NM. Parker VT.017 Collecting and measuring standards of life-history traits of the Jow WM. Adriaens D. doi:10.x TD. Nadrowski K.1046/j. Pausas JG. Klotz S. Tackenberg O. predicts sapling leaf lifespan and shade tolerance of tropical tree Quétier F. e42604. Ford H. Messie J. Rammig A. Violle C. Wirth C (2011) TRY – a global doi:10. Nöllert S. but not lamina Lavorel S. Zanne AE.2307/2446360 Casanoves F. Yamada T. American Journal of Botany 85. Bullock SH. K Thompson) pp. Pillar tropical rain forest. Perspectives in Plant Ecology. Evolution Westoby M. Lambers H.1016/S0065-2504(08)60148-8 database of plant traits. Journal of Archaeological Science 37. Flores O. RM Bekker. Siefert A.2005. Kleyer to maximum height in a Bornean mixed dipterocarp forest. Bossuyt B. Westoby M (1998) Seed size and shape are not related to GM. 940–946.2003. Steendam HJ. Jansen S. Chambers JQ.002 Van Bodegom PM. Soudzilovskaia NA. Cornelissen JHC. Adenostoma fasciculatum (Rosaceae). patterns as correlates of seedling shade tolerance of 13 tropical trees. 66–88 (University of Groningen: Groningen. Knevel IC. doi:10.02451. Van Groenendael JM.1111/j.x B (2008) The LEDA traitbase: a database of life-history traits of the Jones G.jas.1365-2435. (Eds H De Kroon. Salgado-Negret B. Fang J.2008.1365-2745. Jennings DH (1976) The effects of sodium chloride on higher plants. (Eds IC Knevel. Journal of Ecology 96.1111/j. Waldram M. Joly CA. Klimešová J. doi:10. Pérez-Harguindeguy N. In ‘The LEDA traitbase. species from a broad range of habitats.1111/j. Klimeš L. Annals of Botany 104. Craine JM.1469-8137.1998. McIntyre S.03212. Finnegan PM.10.00810. Kitajima K. Lavorel S. McIntyre S.226 Australian Journal of Botany N. Sage RF (2003) Photosynthetic pathway alters xylem structure Ma S.2307/2442650 The Netherlands) Juneau KJ. Poorter L. Ordoñez J. Grigulis K. depend on leaf traits but not on nitrogen supply. Charles M. patterns of seed reserve Pausas JG. Knops JMH. Lens F.2307/3235647 Reinsch S.foreco. Chave J. Kitajima K (1996) Cotyledon functional morphology. doi:10.1017/S0266467499000723 regeneration – A literature review and proposal for simple Kattge J.111. thickness. Jalili A. M Stadler. Lee Ecology 91. Enquist BJ. Kubo T (2003) Tree Gallagher RV. (Cambridge University Press: Cambridge. Kühn I. doi:10.066 and ecosystem functioning from plant traits: revisiting the Holy Grail. Lenz T.x Fidelis A. Roumet C. Advances in Ecological Research 23. Weiher E. American Journal of Botany 67. Cornwell WK. Sardans J. PLoS ONE 7. Prentice IC. Shiodera S. 1–29 (Backhuys Publishers: Leiden.x .x Ogaya R. Thomsen K (1994) A new method for measuring tree height in Ozinga WA. Kramer K. Duarte L. Harrison SP. Korning J.1365-2486. related to adult tree size in a Malaysian mixed dipterocarp forest. D Kunzmann. Bond WJ. Dannemann A.tb01064. Klimešová J. Journal of Vegetation Science 5. King DA. Blonder B.2010. Dobrin BH. doi:10.1016/j. Reu B. Patiño S. Bekker RM.1016/S0531-5565(00)00233-3 Forest Ecology and Management 223. Leadley P.1111/j. 193–210 ecosystems in a changing world’. 419–428. Baker TR.pone. Klimeš L (2007) Bud banks and their role in vegetative doi:10. Kitajima K (1994) Relative importance of photosynthetic traits and allocation Functional Ecology 16.x methods. Phillips OL. Lloyd J. Kummerow J (1980) Leaf turnover rates of northwest European flora’.1654-1103. Kunzmann Ecology 12. Oleksyn J. Biological Reviews of the Cambridge Philosophical Society 51. Williams NSG. Pintor C. Pearse SJ. UK) Poschlod P. Estiarte M. fire and logging in Paraguay and Netherlands) Venezuela. Harrison SP. Anand M. Poschlod P.tb02188.1111/j. Sack L. 797–806. Global Change Biology 17. Garnier E. Garnier E. European flora: the LEDA database. In ‘The functional types: are we getting any closer to the holy grail? In ‘Terrestrial ecology of tropical forest tree seedlings’. Klimešová J. Laliberté E. Noor NSM (2006) Growth and mortality are Experimental Gerontology 36. doi:10. Ryan MH.1365-2478. Moles and hydraulic function in herbaceous plants. White M. Elser J.2006. New Phytologist 186. doi:10. Durka W. classification and assessment. Van Groenendael JM (1997) permanent and temporary storage. Bogaard A. Reichstein M. doi:10. Garnier E. Higgins SI. Partomihardjo T. Hickler T. Llusià J. Journal of Tropical Ecology 15.174318 1151–1161. Thornton P. and Systematics 8. The sprouts after slash-and-burn agriculture.01430.08. Carter GA (1998) Variability in leaf optical properties among 26 Ackerly DD. Boedeltje G. Thompson K. 256–261. 26.2003. Hodgson J (2010) Crops and weeds: the role northwest European flora. Manning P. Cavender-Bares J. Tarasoff CS (2012) Leaf area and water content changes after Klimeš L. Cornelissen JHC. Journal of M. Reich PB. Simpson RL (2008) ‘Seedling ecology and evolution. Urcelay C (2007) Plant utilization and regeneration niches of tropical tree seedlings. Wright IJ.’ Kleyer M.x Oecologia 98. (Ed. Thompson K. D. the field – methodology matters! Functional Ecology 22. Yguel B. Römermann C.1111/j.1046/j. Klimeš L. Díaz S.1016/j. Leishman M. 152–158. 2905–2935. Esser G. Louault F. 611–614. Mahecha MD. Pérez-Harguindeguy et al.1371/journal. Bradstock R. Prinzing A. Hendriks RJJ. Bekker RM. Dorrough SB. Zaehle S. Kerkhoff AJ. Gutierrez AG. Hodgson JG. Gimenez O. Knevel IC.1007/BF00324232 Lavorel S. Pitelka L) (UNESCO: Paris) pp. Bönisch G. Stadler M. 143–157. Fagan WF. doi:10. 1266–1274. doi:10. V. Niinemets Ü. Müller SC. MD Swaine) pp. 1058–1066. Kühn I. Kammesheidt L (1999) Forest recovery by root suckers and aboveground JM Van Groenendael) pp. Clonal plant architecture: a comparative analysis of form and function. Garden D. 545–556. Onoda Y.2011. 453–486. Frank D. Proulx R. Thébault A. Peco doi:10. 115–129. Rusch Leishman MR. Sosinski E.ppees. Cell & Environment AT. Klotz S. Wilmann B. Overbeck G.x Veneklaas EJ (2011) Phosphorus nutrition of proteaceae in severely Kazakou E. Journal of Vegetation Science 14. 70–77. Plant. Bunker DE. Lambers H. Hodgson species differentiation in growth. Green WA. Sonnenschein M. Lewis SL. Bahn M. Finegan B.2002. Poorter L (2010) Tissue-level leaf toughness.x Leck MA.2003. Ozinga WA. Soussana J-F. Swaine E. Götzenberger L. Fernández-Méndez F. Shane MW. Davies SJ. Peñuelas J. Schlegelmilch J. Nüske A.00215. PD Canadell JG. Knapp AK. doi:10. of weed functional ecology in the identification of crop husbandry doi:10. Functional Endels P.1469-185X. Swenson N.01119. Suzuki E. Lavorel S. 480–485. Garnier E (2009) Litter phosphorus-impoverished soils: are there lessons to be learned for quality and decomposability of species from a Mediterranean succession future crops? Plant Physiology 156.00664. Bonis A (2008) Assessing functional diversity in species.10. Chapin III FS.1365-2745. 139–140. Poorter H (1992) Inherent variation in growth rate between higher Shipley B. 2015–2026. Laughlin D. Onipchenko VG. 651–673. Garnier E (2002) Predicting changes in community composition doi:10. 187–261. Violle C. Bakker JP.1976.1046/j. Díaz S. Bekker R. (Ed. Roumet C. persistence in soil in Australia in the same way as in Britain. 134–147. Massad T. Cornelissen JHC. doi:10. Kohyama T. Jackel A-K. Kurokawa H.1016/j. recruitment and allometry in relation JG. Poorter H.0042604 In ‘The ecology and evolution of clonal plants’. Naeem S. doi:10. Paula S. Kirkup D.

Environmental ecosystems. density and thickness in trees and shrubs.x Mooney HA (2010) Biodiversity targets after 2010 Current Opinion in Moles A. doi:10. doi:10.14194.1111/j.’ 4th edn.00680. Westoby M (2000) The evolutionary ecology McIntyre S. 153–175. Ecology 85.x Meinzer FC. steps in the analysis of attribute response and plant functional 2nd edn. Cramer W.1073/pnas. 423–433.1098/rstb. mediate selection by mammals.2010. Perrings C.2307/2401670 Magnani F. Walpole M. Weiher E. Westoby M (2006) Seed size and plant strategy across the Mace GM.02. Goldstein G (2001) Water transport in trees: Litton CM. 91–105. Maherali H. 10540–10544. Oecologia 168.2000.2307/1933631 can adjust hydraulically to their environment. Ackerly D.2006. R Lal) pp. doi:10.1098/rspb.1365-2745. Plant. Phillips N. doi:10. Journal of Ecology 97.2006. 671–677.2307/2937284 Newman EI (1966) A method of estimating the total root length of a sample. Clearwater MJ.1890/0012-9658(2001)082[0453:GSCCOL]2.2000.00991.1399-3054.002 doi:10. Raich JW. Jackson RB (2000) A universal molecular method for forms.tree. Pockman WT.1006/anbo.01526. 2184–2199. 1046–1051.x doi:10. MH ratio. erosion research methods’.x Mäkelä A. Vellend M. Hubbard R. Schachtman DP. 279–287. Wright IJ. and Mazer J (1989) Ecological. Rivelli AR. 31–57 (CAB International: London) types. Proceedings of the Maherali H. doi:10. ecological scales? A case for trait-based ecology. 3–8. Leishman MR (2009) Global patterns in plant vulnerability of woody plants to xylem cavitation. 12–20. traits. Enquist BJ. new insights and some controversies.2006.CO. doi:10. Faith DP. Westoby M (2006) Rebuilding community Niinemets Ü (2001) Global-scale climatic controls of leaf dry mass per area. London) Lagudah ES. Cell & Environment 26. height. Hemmings FA. Journal of Experimental Myers J. M Fenner) pp. M. Barnard H. Ezcurra E (2007) The narrow-leaf syndrome: a functional and salt tolerance of crops.2009. doi:10. 913–920. Lindsay MP. FL) Lüttge (1997) ‘Physiolgical ecology of tropical plants.’ (Springer-Verlag: Milla R. Biological Sciences 334. Hinckley T. 385–392. Díaz S. doi:10. doi:10. Westoby M (2006) Interrelations among pressure-volume doi:10.1111/j. 2109–2115. Ecology Letters 13. toughness of the leaf of the dicotyledon Calophyllum inophyllum L 163–182. Molecular Ecology 9. James RA. Dominy NJ. Tavsanoglu Ç. Philosophical Transactions of the Royal Society of Messier J. 239–262.’ (Academic Press: Munns R. Hare RA (2002) Avenues for increasing Martorell C.1461-0248. USA 102. UK) McGill BJ. DeLucia EH (2001) Influence of climate-driven shifts in biomass National Academy of Sciences. 139–145.0417 of leaf traits responsible for seasonal and inter-annual variability in Milton SJ (1991) Plant spinescence in arid southern Africa – does moisture ecosystem CO2 exchange? Functional Ecology 25. 35–44. 258–270. Oecologia doi:10.1007/s00442-006-0614-x Mutch RW (1970) Wildland fires and ecosystems – A hypothesis.1016/S0098-8472(01)00074-0 Loehle C (1988) Tree life histories: the role of defenses.0501473102 129. In (2002) The relationship between tree height and leaf area: sapwood area ‘Tropical trees as living systems’.003 (2005) Factors that shape seed mass evolution. 561–573. 481–491.1016/j. In ‘The ecology of regeneration in plant communities’. Tweddle J. Reich PB (2007) The scaling of leaf area and mass: the cost of light Berlin) interception increases with leaf size. Fernández RJ (2006) Stay wet or else: three ways in which plants 51.2007. Yamashita N (2000) Mechanical defences Meyer LD (1994) Rainfall simulators for soil conservation research. Physiologia McIntyre S.1139/x88-032 transport: short-term regulation. (Eds PB Tomlinson.2010. Lechowicz MJ (2010) How do traits vary across London. Larigauderie A. ecology from functional traits. Husain S. and the role of physiologically based selection evolutionary approach to the form of fog-harvesting rosette plants. 3963–3977. Meinzer F. 453–469.1365-3040.1046/j.1111/j. Proceedings. Canadian Journal Mencuccini M (2003) The ecological significance of long-distance water of Forest Research 18. McDowell N. In ‘Handbook of plant functional ecology’. doi:10.1991. allocation on water transport and storage in ponderosa pine.00535. Lenz TI. Dickie J. Turner IM. Journal of Ecology 89. In ‘Soil to herbivory. Pausas JG (2012) Local versus regional different age and size classes. Biological Sciences Ma S. Zanne AE.1007/s00442-002-0904-x Zimmerman) pp. Scholes RJ. invasion. Köstner B.1111/j. Berrick AJ (1991) The fracture costs of stature across plant life forms. Hoffman GJ (1977) Crop salt tolerance – current assessment. Wright I.2001. 407–448 (Marcel Dekker: New York) 1549–1559.1093/jxb/erl127 tailed deer: implications for long-distance dispersal. and Experimental Botany 45.1046/j.x Lucas PW.1365-2435. Ecology 82.1365-2745. doi:10.1007/s004680100118 doi:10. Annals of Botany 86. Marshall J. Forbes TDA (1999) Disturbance Plantarum 127. Series B.0030-1299. Dawson TE (2011) Are temporal variations 274. Ryan MG (2007) Carbon allocation in forest current perspectives.1023/A:1021119414799 Oecologia 151.2307/3237077 Press: New York) Medina E (1999) Tropical forests: diversity and function of dominant life- Linder CR. Bond B.2007. doi:10. Journal of Applied Ecology 3.1365-2486. Warton DI. McGill BJ. Oecologia 139. 923–932. Moles A. Baldocchi DD. Laffan SW. Warman L. doi:10. Lavorel S.2003.1007/s00442-011-2127-5 Marschner H (2012) ‘Mineral nutrition of higher plants. Moles AT. Swenson NG. Marks P (2004) Seed dispersal by white- Botany 57. 95–106. Schopfer P (1995) ‘Plant physiology.0099 838–848. Whitehead D Ng FSP (1978) Strategies of establishment in Malayan forest trees. 2089–2109. Mohr H. Watson JEM. Mambelli S.03.x doi:10. Trends in Ecology & Evolution 21. doi:10.01476. (Ed. and life history correlates of seed mass migration of plants in eastern North America. identifying underground plant parts to species. doi:10. Walther BA. 93–105. doi:10. Oecologia 132. Palmer whole life cycle Oikos 113.’ (Academic traits. Ryan M. Journal of Vegetation Science 10.1046/j. Ecology Maseda PH. Plant and Soil 247. among Indiana dune angiosperms. Trees 15.cosust.2 . Lavorel S (2001) Livestock grazing in subtropical pastures: of seed size. Gardescu S. F Valladares) pp. doi:10. Landsberg J.01779.x response in vebetation – towards a global perspective on functional Levitt J (1980) ‘Responses of plants to environmental stresses.1890/02-0538 doi:10.1007/s00442-003-1474-2 doi:10. Vanninen P (2001) Vertical structure of Scots pine crowns in Moreira B.1111/j. Tan HTW. 209–226. taxonomic. doi:10. Jackson RB (2004) Adaptive variation in the Pitman A. doi:10. Turner IM. doi:10. Pitman A. Oecologia 87.1111/j. 115–134.1016/j. doi:10.New handbook for measurement of plant traits Australian Journal of Botany 227 Leishman M. Moore LA. Choong MF.x (Guttiferae). Global Change Biology 13.1007/BF00325267 Maas EV. Le Prestre P. Ecological Monographs 59.0. (Ed. long-term acclimation and the hydraulic Lucas PW. Westoby M Environmental Sustainability 2. doi:10.2010.1261 Beach.01420.1365-294x. (Eds FI Pugnaire. 129–162 (Cambridge University Press: Cambridge. Condon AG. 621–630. 178–185. Ishii H. 209–222. Webb C. intraspecific variability in regeneration traits. 83–104 (St Lucie Press: Delray doi:10. Moles A.x curve traits across species and water availability gradients. (Springer: Berlin) Journal of the Irrigation and Drainage Division 103.01034.

727–736. Choong AMF. Moreno JM. Gleichman JM.2011. Cobb ED. doi:10.x physiological and structural traits in Amazon forest trees.1461-0248.1890/08-1309.x doi:10. 87–95. Proceedings of the National Academy of Sciences. south-east Australia. Leal E. Shipley B. Lloyd J. doi:10.2002. 283–303.tb01053. Yamashita N (2011) Cabido M (2003) Leaf traits and herbivore selection in the field and in Global patterns of leaf mechanical properties. Fernandes PM. Santos AJB. Martin ME. New Phytologist 156. F Valladares) pp. Ecology Letters 12.8-9. Higuchi N. Trabaud L (1990) Structural characteristics of fuel components of Oecologia 153. Witte J-PM. 81–120 (Marcel Dekker: New York) . Noble PS (1992) Drought-induced changes in hydraulic doi:10. Makarov MI. Seed A. Richards L. Palomino W. Elgart A. Teekev DK. Pickett STA. 249–259. conceptual approach.x Onoda Y.2009. Pausas JG (2006) Leaf traits and resprouting ability in the Olff H. Peña-Cruz A. Priante Filho N. Markesteijn L. Santiago LS. Valladares F Parton W.1111/j.1134853 doi:10. Paula S.5194/bg-6-545-2009 conductivity and structure in roots of Ferocactus acanthodes and Patiño S. doi:10. Gallo J. Fasth B (2007) Global-scale economics? Disproportional scaling of support costs within the similarities in nitrogen release patterns during long-term decomposition. Adler PB. carbon assimilation. Phillips OL. Díaz S. McArthur C. 9–19. Loucks OL (1972) Optimal leaf size in relation to environment.03945. of Vegetation Science 8. Garnier E (1999) Ecological significance of relative growth rate Parkhurst DF.1992. Forest Ecology and Management 35. Van Bael SA. Bokdam J. Alessio GA. Quested HM (2007) Sampling plant functional traits: what daily CAM use by Bromeliaceae.1111/j. Pausas JG (2009) Fire-related and boreal forests: functional relations and potential climate feedbacks.1442-9993. 1420. New Phytologist 193. Peacock J. Slatyer RO (1980) The use of vital attributes to predict successional Salomão R. Austral Ecology 28. Harmon ME. Brandt LA.1016/0378-1127(90)90006-W performance across 53 rain forest species. 758–764. 8891–8896. wet environments: does theory mirror nature? American Naturalist 175. Palacio S. Wright IJ C. Mercado LM. Sosinski EE. Sarmiento Niklas KJ.589 Pons J. New Phytologist 86. doi:10.01321. Paiva R. Science 315. Matesanz S. 5–21. Herrera R. 642–650.1469-8137. Schieving F. Chave J. JHC. Czimczik CI. Pausas JG. IL) Hoyos EJ. Functional Ecology 20. doi:10. spectrum of leaf physiognomy. Silva N. and albedo in temperate Fernandez-Santos B. Alvarez E.CO.1980. doi:10. Fyllas NM. doi:10. Malhi Y. Kazanis Tavsanoglu Lloret F. USA 104. Schmerler J.tb00507. Hart SC. USA 105. doi:10.x Ordoñez JC. Soares de Almeida S. 703–706. Global Ecology and Biogeography 16. Arianoutsou M. Pitman N. 321–331. Lloyd J (2012) Coordination of doi:10. Pau U KT. Traba J. Díaz S. 301–312.2010.1111/j.5194/bg-9-775-2012 20.1890/0012-9658(2006)87[1733:LTAGPO]2. Annals of Botany 101. doi:10.2006. Joseph Wright S. 91–96. Burke IC. Gurvich DE. Vera FWM. (2007) Do we underestimate the importance of leaf size in plant King JY. Westoby M. 505–537. Bongers F (2006) Leaf traits are good predictors of plant doi:10.1007/s00442-007-0788-x five Mediterraean shrubs.x Pisani JM. Van der Willigen C (1998) A mathematical and statistical of spines and phenols in Prosopis caldenia and Prosopis flexuosa. In ‘Handbook of functional plant ecology’. doi:10. Smit R (1999) Shifting mosaics in grazed woodlands driven by the doi:10.and intraspecific variations in production Pammenter NW. Plant Biology 1. Wright IJ. Van Logtestijn RSP. Bonal D. doi:10. Journal Onipchenko VG. Howard JJ.0.2006. Frolking SE. Baker TR. Distel RA (1998) Inter. Núñez-Vargas P. changes in plant communities subject to recurrent disturbances.1469-8137.00283. doi:10.tb04244.1093/treephys/18.1111/j.x 225–239. Parkhurst DF (1994) Diffusion of CO2 and other gases inside leaves.1007/BF00121013 density variations across the Amazon Basin. Lucas PW.1007/s00442-010-1806-y PB.1111/j.’ (The Schwarz M. cafeteria experiments. Monteagudo A. doi:10.1111/j. 75–83. Quesada CA. Niinemets U.1086/649582 Pierce S. (2007) “Diminishing returns” in the scaling of functional leaf traits across Baraloto C. Quesada CA.00489. Sellin A. Paula S. Phillips OL (2009) Branch xylem 43. de Maeyer K. University of Chicago Press: Chicago. 18–23.1461-0248. Noble IR.1016/0031-9422(81)85134-5 Paula S. Katul GG. Van Hal JR.x alternation of plant facilitation and competition. Day MC. New Phytologist 120. doi:10. Sánchez AM. Schwarz M. Kempf JS (1980) Branching patterns in forest shrubs and Aerts R (2010) Plant strategies in relation to resource supply in mesic to understory trees in relation to habitat.2307/2258359 Pugnaire.x between resprouting life histories. Science Research 13. Dominy NJ. Cornelissen JHC. Sota A. Tena D. Ecology 87. Poorter L.x doi:10. Arroyo L. Mediterranean basin. Opuntia ficus-indica.1438-8677. Sciences. Plourde LC. Martin TA.1023/A:1022380627261 Physiology 18. Ecology Letters 14. (Eds FI Journal of Ecology 60. Buhk C. Paiva R. Baker TR. doi:10. Tree Journal of Chemical Ecology 24. Currie WS. Niklas KJ (1994) ‘Plant allometry: the scaling of form and process. Phytochemistry 9. Vegetatio Vásquez-Martínez R. Cornelissen JHC (2009) New nitrogen uptake productivity and disturbance gradient in mediterranean shrublands of strategy: specialized snow roots. 1733–1743. Tobias M. Adair EC. Biogeosciences 6. (2008) Canopy nitrogen. Clissold FJ. Monson RK. doi:10.1046/j. Horna V. Aguilar A.1093/aob/mcm107 Patiño S. ter Steege H. 330–340.2003. Hollinger DY. Vieira I. Smith M-L. 23–36. Cook BD. Cornelissen Peco B. Bakker ES. Rodrigo A. Silver WL. Ermak AA.1469-8137. doi:10.x Science 10.1073/pnas.228 Australian Journal of Botany N. Neill D. Bolstad PV. 361–364.1111/j. Bradstock RA (2007) Fire persistence traits of plants along a Kozhevnikova AD.x Poorter H. Killeen and within species groups.1111/j. proportion of the species need to be measured? Applied Vegetation doi:10. Silveira M.2307/3237375 Akhmetzanova AA.1466-8238. Meir P. Levassor C. New doi:10. Pausas JG (2007) Acorn dispersal estimated by radio-tracking Papió C. Pérez-Harguindeguy N. Rudas A. Anten NPR (2008) Effects of light and nutrient Pausas JG.01185. 553–567.tb00236.1046/j. Prieto A. Ecology 90. Vendramini F. Oren R. Kitajima K. Ivanov VB.1126/science. doi:10. 775–801. Luizão F.01331.0701135104 Peñuela MC. Biogeosciences O’Leary MH (1981) Carbon isotope fractionation in plants. Richardson AD. 941–947. Van Bodegom PM. Kurokawa H.1365-2435. Pausas JG (2011) Root traits explain different foraging strategies 127–137. Munger JW. Turriago JD. Bartholomeus RP. Ollinger SV. Schmid HP Ojeda F. Niinemets Ü. doi:10.x Onoda Y. Poorter L. Moreira B. Panfil SN. 449–479. Griffiths H (2002) Carbon isotope ratio and the extent of Pakeman RJ. Azcárate FM (2003) Seed size. Luna B. Proceedings of the National Academy of T. and its components. Oecologia 165. Jalili shape and persistence in dry Mediterranean grass and scrublands. Enrico L. Corcobado G (2012) Fires enhance availability on leaf mechanical properties of Plantago major: a flammability in Ulex parviflorus. 589–593. Jimenez EM. North GB.0810021105 Pausas JG (1997) Resprouting of Quercus suber in NE Spain after fire. 545–568. Salpargarova FS. Villanueva B.1469-8137. doi:10. Warton DI. Vitzthum P.1 19 336–19 341.1999. doi:10.1469-8137. Grassens L. Torres-Lezama A. Espelta JM. Annals of Botany 100.1111/j. Santos AJB. Portsmuth A. Fine PVA. Costa ACL.2007.1079/SSR2002127 Pérez-Harguindeguy N. 903–911. traits for plant species of the Mediterranean Basin.1073/pnas. Reich PB.1111/j.1654-109X.01582. analysis of the curves illustrating vulnerability of xylem to cavitation. Pérez-Harguindeguy et al. Reich doi:10. Winter K.2 Phytologist 126. 219–228.1994.tb03191.

2307/1479087 tolerance in seedlings of nine boreal tree species grown in high and low Poschlod P. Oecologia 158. Kiefer S.CO. Plant Physiology 155. (Ed. Bonn S (2005) Plant dispersal potential and its Bowman WD (1999) Generality of leaf trait relationships: a test across six relation to species frequency and coexistence. Vanderklein DW.1007/BF00317383 Poorter H. stems and roots: meta-analyses of global convergence in plant functioning. Paz H. the relationships between leaf nitrogen. Read J. Craine J. doi:10. American Journal of Botany 90. Mommer L (2012) Tansley Reich PB. Wright SJ. Hedin L. plant and stand characteristics among diverse ecosystems. Biological Sciences 277.1991. 1011–1020. Bushena C (1998) calcareous grasslands as affected by dispersability in space and time. E152–E161. Symonova O. 1289–1301. Montalvo A. (2002) Fine root architecture of nine North American trees. Gunin PD. 10 101–10 106. Niinemets Ü.1365-3040. Ruess RW. 30–50. nutrients and water: a Raunkiaer C (1934) ‘The life forms of plants and statistical plant geography. Ecology 87.1600-0706. pp.x Resprouting as a persistence strategy of tropical forest trees: its relation Reich PB.1104/pp. Villar R (2009) Causes and Reich PB. Physiologia Plantarum Rabotnov TA (1950) The life cycle of perennial herbaceous plants in meadow 67. doi:10. Prins HHT (2010) 251–259. Melgar I. Bongers L.00208.2 Pregitzer KS. New doi:10.2307/2937116 Poorter H. Allen MF. Oikos 98. Phyllis DC.1071/PP99173_CO American Journal of Botany 93. leaf and root morphology. Knops J. Trudy Botanicheskogo Instituta Akademii Nauk SSSR Seriia III Poorter H. Lugo AE (1992) Wood densities of tropical snapping of trees: structural determinants and ecological consequences. 107–110 (Chapman (2010) Partitioning the components of relative growth rate: how important and Hall: London) is plant size variation? American Naturalist 176.1998. influence biomass accumulation and its response to CO2 and N. Walters MB.02830. shoot allometry in contrasting climates. determinant of leaf structure and function among 23 Amazonian tree Ecology 89. Ackerly DD (2003) Hydraulic architecture and the evolution of Proceedings of the National Academy of Sciences. doi:10.1469-8137. 793–801. 2613–2627. USA 101. doi:10. half’. Gresham C. 16–24. Oleksyn J. Tackenberg O (2000) Databases on life history traits as light. Ellsworth DS.1086/657037 species: traits. Bongers F (2006) Architecture of 54 moist-forest tree doi:10. Garnier E (2007) The ecological significance of variation in relative Pyankov VI.3732/ajb. Walters MB. Tilman D. 1955–1969. Oecologia 86.3732/ajb. U Kafkafi) pp. Ellsworth DS (1997) From tropics to tundra: review.2002. Analyzing variation in RGR in terms of the doi:10. Noy-Meir I. a tool for risk assessment in plant species. F Valladares) pp. 15–31. doi:10.x photosynthesis in maple and oak trees. Walters MB. New Phytologist 193. pp.tb02447.0306602101 1502–1512.1111/j.1111/j.162834 Reich PB. Mercado P. Volin JC.94. Pugnaire FI (1999) Plant survival in arid environments. 365–392. 314–338 (Marcel Dekker: Harms KE.1111/j. F Valladares) major plant groups and biomes. doi:10. Biomass allocation to leaves. General technical report S0-88 US Department of Canadian Journal of Forest Research 13.10. Tjoelker MG. Functional Ecology 12. Turnbull LA. Osborne CP. 35–46. Proceedings.’ quantitative review [published erratum appears in Australian Journal of (Clarendon Press: Oxford. Mazer SJ.1502 Reich PB.1365-2435. doi:10. Oleksyn J. Chapman J. Wedin D. stems and roots of and analysis framework graphical user interface: segmenting and higher land plants.2009. Hendrick RL Reich PB. In ‘Vegetation ecology’. doi:10. 1546–1565. Lu K. Uhl C.13730 Poschlod P. Forest Service. Reich PB. UK) doi:10.93. Preston KA. Zeitschrift für Ökologie und doi:10. 1191]. 75–91. Bonn S (1998) Plant species richness in Reich PB. In ‘Methods in comparative plant ecology – a Rees M. (Eds GAF Hendry. USA 94. Condit R. Ackerly DD. 1908–1920. (Eds FI vegetation of Mongolia: their natural occurrence and geographical Pugnaire. Ellsworth DS (1991b) Leaf age and season influence consequences of variation in leaf mass per area (LMA): a meta-analysis. trade-oofs.1986. Tsoog S. Martínez-Ramos M. Taylor SH laboratory manual’. Poorter H. Stokes A (2006) Plant biomechanics in an ecological context.0. Poot P. Chubiña J. Mileyko Y. Reich PB. Aiello A (1983) Uprooting and Reyes G. Close association of RGR. Black CC (2000) C-4 plants in the growth rate and its components. (Eds Y Waisel.CO.2 adaptation or the ‘Emperor’s new clothes? In ‘Plant roots: the hidden Poorter L.1 Monographs 62. Ecology Letters 11.x underlying carbon economy. .1111/j. Wright IJ. doi:10. Ellsworth DS (1992) Leaf life-span in relation to leaf. Tjoelker MG. Chapman PL (2002) The role tropical tree species.01185. Southern Forest Experiment Station. doi:10.x Naturschutz 9. Machado JL Price CA. seed mass and shade Applied Vegetation Science 1. A Eshel. Nagel O (2000) The role of biomass allocation in the growth 6. Burton AJ. Vose JM.1111/j.x coenoses.1890/0012-9658(2006)87[1289:AOMTST]2. Oecologia 123. Poorter L. Plant. New York) Peña-Claros M. Milchunas DG. Tränkle U. Niklas KJ. Hulme SP. 3–18. 53–64. UK) Plant Physiology 27.1 species. 13730–13734. Woodward FI. leaf mass per area and New Phytologist 182. FL) distribution in relation to climate. Ecology 80.1469-8137.1007/s004420050985 a method avoiding curve fitting and pairing. and functional groups. doi:10. 381–405 (Marcel Dekker: New York) 877–883. with carbohydrate storage and shade tolerance. Agriculture. JP Grime) pp.1890/0012-9658(1999)080[1955:GOLTRA]2. Wright IJ (2008) Are functional traits good Reich PB. E Van der Maarel). 565–588. analyzing the structure of leaf veins and areoles.1546 Poorter H.2009. Pregitzer KS.03952. Elser J (2010) Evidence Puigdefábregas J.10. Licona JC. Niklas KJ.x 236–245.1046/j.980106. response of plants to different levels of light. Brown S. DeForest JL.1034/j. LA. Kleyer M. 223–226. (Eds FI Pugnaire.x doi:10.2011. Muller-Landau HC. tree species. Ecology 72. doi:10. Proceedings of the National interspecific variation and environmental control. Kitajima K. Wright IJ. Tackenberg O.1098/rspb.25.New handbook for measurement of plant traits Australian Journal of Botany 229 Poorter H.1073/pnas. Walters MB. Reich PB (2002) Root-shoot relations: optimality in acclimation and doi:10. biomes.1461-0248. CO2.1139/x83-133 Orleans.1818 Putz FE. Poschlod P. Weitz J (2011) Leaf extraction (2008) Scaling of respiration to nitrogen in leaves. Australian Journal of Plant Physiology 27.0. Ibarra-Manríquez G. Rozendaal DMA (2008) Leaf size and leaf display of thirty-eight Rebollo S. Trost J (2004) Species and functional group diversity independently 293–309. 327–338. Walters MB. Wright IJ. 147–171 (Blackwell: Oxford. In ‘Functional plant ecology’.1399-3054.tb01499. doi:10. Welschen RAM (1993). Hilley T. of a spiny plant refuge in structuring grazed shortgrass steppe doi:10. Walters MB. 67–100 (CRC press: Boca Raton.1073/pnas. doi:10. 7–204. doi:10. Poorter H. Ellsworth DS.1890/07-0207. Academy of Sciences. Poorter L.2008. 595–607. Webb CO.1007/s00442-008-1131-x plant communities. Poorter L. Cell & Environment 14. Oleksyn J. Lewis C (1986) Testing differences in relative growth rate: doi:10. Ellsworth DS (1991a) Leaf lifespan as a predictors of demographics rates? Evidence from five neotropical forests. Ecological doi:10.110.1890/09-0862. Naeem S. Ecology 91. In of a general 2/3-power law of scaling leaf nitrogen to phosphorus among ‘Handbook of functional plant ecology’.90.

359–364. Switzerland) Roth-Nebelsick A.1111/j. Hoogedoorn H. Bond WJ. Evans JR.00320. 159–162. life Sack L.1600-0706. Buizer B. Kerp H (2001) Evolution and Schweingruber FH. D Kunzmann. Rajabi A. age determination and stem anatomy. Tackenberg O. doi:10. Oecologia 70. 808–817.1407 (Groningen University: Groningen. Robson TM. 312–319. Biology 57.x doi:10. Yaqine Shipley B.1111/j. Weijers S. doi:10.1146/annurev. orientation.01667. Sack L. Forest Snow and Landscape 553–566. In ‘The Scholander PF (1966) The role of solvent pressure in osmotic systems. Functional Ecology 20. traits. Prast G. 357–368.1007/BF00044777 doi:10.032604. D Kunzmann.1073/pnas. Functional Plant Biology conductivity and embolism in xylem.0. 89–114.1046/j. Vegetatio 62.13911. Annals of Botany 87. Jackson RB (2002) The global biogeography of roots. doi:10. 543–550. Díaz S (2006) Suites of root traits differ between Seibt U. Oecologia 155.2001. Woodruff D. 361–381. Atkin OK. 483–491. 195–415. 37. Tree Physiology 30. Tackenberg O. Mayoral Fuertes M. Plant Biology 3.1365-2435.x 51–64. Fisher JB. Broekman R. RM Bekker. 2461–2467.2005.x Thompson) pp. Plant Ecology 168. 1407–1414. K Thompson) pp. 311–328. of Cassiope tetragona as a proxy for Arctic climate: developing doi:10. 441–454. Functional Ecology 9. Sperry JS (2003) Evolution of water transport and xylem structure. Higgins SI (2005) A mechanistic model Römermann C. 214–224.’ (Oxford University Press: Jones T (2004) Leaf photosynthetic traits scale with hydraulic Oxford. 1017–1028.1461-0248. Annual Review of Plant of Animal Ecology 43. DC) doi:10. doi:10. (Eds HA Mooney. doi:10. and sunlight exposure in five Western Australian doi:10. Biology 15.1365-2745. Ecological potential of seeds to sheep and cattle coat from simple morphological seed Monographs 72. McElwain JC. Rodriguez-Calcerrada J. S115–S127. Poschlod P (2005) Growth rings in herbs and shrubs: life function of leaf venation architecture: a review.’ Netherlands) (Haupt-Verlag: Bern. Richter M (1992) Methods of interpreting climatological conditions based on Sage RF (2001) Environmental and evolutionary preconditions for the origin phytomorphological characteristics in the cordilleras of the Neotropics.1469-8137. In ‘The LEDA traitbase collecting and measuring Journal of Ecology 93. WE Winner. 544–548. 337–365. Rotenberg Y. Sack L. Global Change ‘preference’.1111/j. 127–129 (Groningen University: Groningen.1890/0012-9615(2002)072[0311:TGBOR]2.CO.tb01774. (epizoochory). Holbrook NM (2006) Leaf hydraulics.6. 34 species of herbaceous angiosperms.2008. efficiency: sense and sensitivity.x doi:10.01174.1111/j. New Phytologist 153. ‘acceptability’ and ‘electivity’. Tackenberg O. Ackerly DD (2001) Flammability and serotiny as strategies: wildland fuels.2 Römermann C. Merla A (2008) Determination of leaf fresh mass after Science 130.1016/0021-9797(89)90074-X storage between moist paper towels: constraints and reliability of the Smith WK. Meinzer FC.1111/j. Oikos 110. Phytologist 179. Cell & Environment 11.2307/2446554 in tropical rain forest trees.1890/05-0710 Southwood TRE (1977) Habitat. 56–63. New Phytologist 170.1006/anbo.1055/s-2001-15206 Robertson GP. Journal of Colloid and Interface Ryser P.2006. 139–163. The Schweingruber FH (1996) ‘Tree rings and environment.00136.1071/FP10097 35–40. Machado K.1093/treephys/tpp104 doi:10.0030-1299. K doi:10. 687–693. Poschlod P (2005b) Buoyancy. Adams JM. Marañón T (2003) The functional morphology of juvenile expectancy and herbivore damage.x Roumet C.1365-2486. Grubb PJ.arplant.1007/BF00044887 Rozema J.230 Australian Journal of Botany N. Frole K (2006) Leaf structural diversity is related to hydraulic capacity doi:10. doi:10. 1703–1715. 326–336. Uhl D. Gil L.1365-3040.1988. UK) conductivity and wood density in Panamanian forest canopy trees. Service: Washington. Vegetatio 123. Ecology 87.144141 Southwood TRE. USA 55. doi:10. 124–127 doi:10.x Römermann C. structure.1034/j. Midgley GF. Ecology Letters 3. doi:10. plants tolerant of strong summer drought in shaded forest understories in doi:10. (USDA Forest correlated evolution in pines. doi:10. Donnelly JR. the templet for ecological strategies? Journal Sack L.2001. doi:10.2009. Griffiths H. Bledsoe CS. Oecologia 140. ecology and Sperry JS. LEDA traitbase collecting and measuring standards’. Poschlod P (2005c) External animal dispersal for secondary seed dispersal by wind and its experimental validation. Goldstein G.2001. USDA Forest Service Research Paper INT.x doi:10.1111/j.1046/j. Mosbrugger V. Berry JA (2008) Carbon isotopes and water use annual and perennial species growing in the field. Bell DT. doi:10. Journal of Experimental Botany 59. (Eds IC Knevel. Cooper E (2009) Annual growth concentration in plants and their parts.x correlative and experimental transfer functions to reconstruct past Singer MC (2000) Reducing ambiguity in describing plant-insect interactions: summer temperature on a millennial time scale. Neumann AW (1989) Contact angle measurements stresses’. Santiago LS. doi:10. Australian woodlands. Oikos 94. Proceedings of the National Academy of Sciences. M Stadler. 745–752. Werleman C. Munns doi:10.2307/2390579 Rozema J.1007/s00442-004-1624-1 Aranda I (2010) Thermal acclimation of leaf dark respiration of beech Scarff FR.1391 Research 79. doi:10. CA) axisymmetric drop shape analysis. Vu TT (2002) Dry matter content as a measure of dry matter HE. Blokker P. Reader PM (1986) Leaf palatability. doi:10. Shepherd KA (1998) Associations between leaf method.1023/A:1024423820136 International Journal of Plant Sciences 164.2000.01018. Rothermel RC (1972) A mathematical model for predicting fire spread in Schwilk DW. RM Bekker. Brown VK.2006. Bijwaard P. Nicotra A (2010) A unique web resource for physiology. span. M Stadler. doi:10. standards’. Barbour MM.1111/j. New of seedlings of five deciduous broad-leaved tree species.x Rundel PW (1991) Shrub life-forms. methods for long-term ecological research. 202–213.1086/368398 R. Choat B. (Eds IC Knevel.56. Kikuzawa K (1996) Importance of seed size for the establishment and shape to climate within Acer rubrum and Quercus kelloggii. Plant Research and Development 36. Tyree MT (1988) A method for measuring hydraulic the environmental sciences: PrometheusWiki.1469-8137. Wilf P (2008) Sensitivity of leaf size Seiwa K.1007/s00442-007-0932-7 Royer DL.0028-646X. Bernardi J. 345–370 (Academic from the contact diameter of sessile drops by means of a modified Press: San Diego. Urcelay C.1007/BF00379901 southern Spain. The Netherlands) Schurr FM. Sollins P (1999) ‘Standard soil Santiago LS. Coleman DC. 499–521. Plant. 25–34. Zaragoza-Castells J.1093/jxb/ern120 communities. American Journal of Botany 85. Cornwell WK. and diversification of the C4 photosynthetic syndrome.2005.01858. Broekman R (1985) Ecophysiological Shipley B (1995) Structured interspecific determinants of specific leaf area in adaptations of coastal halophytes from foredunes and salt marshes.940213. EJ Pell) pp. Dendroecology. 219–230. Pérez-Harguindeguy et al.x . Poschlod P (2005a) How to predict attachment Schenk HJ. In ‘Response of plants to multiple Skinner FK. doi:10. Westoby M (2006) Leaf litter flammability in some semi-arid seedlings experiencing summer drought in high and low light environments.02496.55.

1469-8137. Jones MH. McCulloh KA (2008a) Safety and efficiency conflicts Tyree MT.4. Available from: http://www.516 Vendramini F. Environment and Quality _Veg. Bakker JP. UK) Walters MB.1469-8137. Poorter L.1046/j. montane tropical rain forest. Hodgson JG.0. Piepho H-P. Trends in Ecology & Evolution 14. 337–361 (Backhuys Publishers: Leiden.2307/3543923 doi:10. Vegetatio 89. Lloret F. Zagt RJ. In ‘Plant life histories’.1007/BF00032170 Taylor B.2000. Annual Review of Plant Physiology and Plant Molecular Environment 31.’ (The Benjamin/Cummings pp. 637–648. Broekhoven AJ.2 in plant growth’. Hendriks RJJ (1997) Swenson NG. Rutherford MC. Kazakou E. Navas M-L.1046/j.1111/j.x Biology 40. 51–64 (European Commission.1 range of species. P Balabanis. R Fantechi) pp. Agrawal AA (1999) The ecology and evolution of plant tolerance life-history variation in a tropical rain forest tree community. Colombia.x University Press: Cambridge.pp. Midgley GF.’ methods for measuring xylem cavitation in conifers. Meinzer FC. Melchinger AE (2008) Multi-trait association Van der Pijl L (1982) ‘Principles of dispersal in higher plants.2007. Wright IJ (2005) Specific leaf area and dry matter content Thomas SC (1996) Asymptotic height as a predictor of growth and allometric estimate thickness in laminar leaves. Houba VJG (2004) ‘Plant analyses procedures.1007/s00122-008-0834-z Van Gelder HA. Sterck FJ (2006) Wood mechanics.1046/j.1999. Poorter H. 23–28. 558–568. Change Biology 7. Band SR. 289–308. II. Klimešova J. (Springer-Verlag: Berlin) porous angiosperms.0.x doi:10. Thompson K.berkeley. Hodgson JG Tackenberg O (2003) Modeling long distance dispersal of plant diaspores by (2002) Leaf traits as indicators of resource-use strategy in floras with wind. Canadian Journal of Botany 66. (Eds O Species composition and fire: non-additive mixture effects on ground fuel Gašparíková.x doi:10. New Phytologist 177. doi:10. 669–675. doi:10. Press: Cambridge. 632–645. Midgley GG. Enquist BJ (2008) The relationship between stem and Comparative ecology of clonal plants. 516–519. Mirotchnick environmental proxy: a critical review and new data. 179–185. 459–471.00063 Stich B. Ecological Monographs 73.40.x (Kluwer Academic Publishers: Dodrecht.2307/2258617 Wilson PJ. Plant. 71–82 (Kluwer flammability. methods of leaf dry matter content produce similar results in a broad doi:10.01795. Bond WJ. Vile D. Cell & embolism.x doi:10. N (2012) Phylogenetic diversity and the functioning of ecosystems. doi:10. doi:10.1365-3040.060189. February EC (2011) History matters: tree establishment Vaieretti MV. Cadotte MW. MacDonald AAM. Lavorel S.1890/ES11-00029. UK) doi:10. 995–1001.x Turner NC (1988) Measurement of plant water status by the pressure Wand SJE.3389/fpls. doi:10. Academic Publishers: Dordrecht. Vile D.2307/2389893 Wahl S.01765. Van Tamm CO (1972) Survival and flowering of some perennial herbs.1016/S0169-5347(98)01576-6 Van Groenendael JM. Jekyll Island.95. Reich PB (1999) Low-light carbon balance and shade tolerance Tjoelker MG. Gurvich DE. In ‘Forest fire and forest meteorology’. GA. Journal of Ecology 92.2002. Van Altena C. 63.00425.’ doi:10.1890/0012-9615(2003)073[0191:AOWDPI]2. 1933–1939. Laurent G. Taneda H.1365-2486.’ (Springer- mapping in sugar beet (Beta vulgaris L. Díaz S. The Netherlands). New to herbivory. Curtis PS (1999) Responses of wild chamber technique.0030-1299.00775. Parkinson D (1988) A new microcosm approach to litter Vesk PA. Roumet C. 236–241. The Genderen M (1990) Hydrological properties of the epiphyte mass of a behaviour of some orchids on permanent plots. The Netherlands) doi:10. Bush SE. 223–230. Temminghoff EEJM. Sperry JS (1989) Vulnerability of xylem to cavitation and in hydraulic architecture: scaling from tissues to trees. Fortunel C. F Eftichidis. Zimmermann MH (2002) ‘Xylem structure and the ascent of sap.CO. Palaeogeography. 556–566. Phytologist 171. EUR 16719 EN’. succulent species. diffuse and ring. Tennant D (1975) A test of a modified line intersect method of estimating root Díaz S. Ecological Monographs 73.2307/2390225 doi:10. density and longevity. 947–954.2012. perceptions. 882–892. Journal of Ecology 63.x Tackenberg O.15559.0022-0477. length.000315 Sperry JS. Reich PB (2001) Modelling respiration of in the seedlings of woody plants: do winter deciduous and broad-leaved vegetation: evidence for a general temperature-dependent Q10 Global evergreen species differ? New Phytologist 143. doi:10. Hodgson JG (1993) Seed size and shape predict seed (2007) Let the concept of trait be functional! Oikos 116. Poschlod P. Annals of Botany 96. Garnier E (2007) Two measurement variability and species turnover in an African savanna. New Phytologist 154. Marushia RG. Theoretical and Applied Verlag: Berlin) Genetics 117.00397. Schulz B. Ecology Letters 15. Frontiers in Plant Science 3.3732/ajb.1469-8137. Bonn S (2003) Assessment of wind dispersal Veneklaas EJ.2001. Poorter L (1998) Growth and carbon partitioning of tropical potential in plant species. 310–320. H Poorter. Oikos 23. tree seedlings in contrasting light environments.00265.1016/S0031-0182(98)00189-8 Staver AC. Van Leerdam A. Gordon DA. Van Ek R. Annals of Botany 99. American Journal of Botany 95. doi:10. In ‘12th conference on fire Valette J-C (1997) Inflammabilities of Mediterranean species. Navas M-L.1046/j. Wilson PJ. Zeiger E (2010) ‘Plant physiology.2307/2445913 Violle C.1469-8137. doi:10.1461-0248.2 doi:10. Palaeoclimatology. I Mistrík. Bekker RM (1997) ‘The soil seed bank of North of grasses.00871. New Phytologist 148.2004.1890/0012-9615(2003)073[0173:MLDOPD]2. 19–36. The Netherlands) Publishing: Redwood City.00357. of Life: Brussels) Steudle E (2001) Water uptake by plant roots: an integration of views. In ‘Inherent variation doi:10. and Strauss SY. Shipley B. American Journal of Botany doi:10.2007. Uhl D.pdf [Verified 22 February 2013]. Vile D.1999.1111/j. J Silvertown. 15–26.1146/annurev. 143–154. doi:10. Westoby M (2004) Sprouting ability across diverse disturbances and decomposition studies. West Europe: methodology. Irrigation Science 9. (Eds H Lambers.1093/aob/mcm022 domestic vegetation subjected to desiccation. Stephens SL. vegetation types worldwide. risk and management. 183–192.01757.1093/aob/mci264 83.x doi:10. Oleksyn J.’ (Cambridge doi:10.x . Garnier E Thompson K. Cornelissen JHC (2012) In ‘Recent advances of plant root structure and function’.CO. 367–378. Hummel I. (Eds JL branch specific gravity and the ability of each measure to predict Harper. Martin RE (1994) Combustibility of selected doi:10. F Baluška) pp.1111/j. Ecosphere 2. 191–209 (Cambridge University leaf area. 173–189. 955–958. art49. characteristics in Malaysian rain forest trees. Van Logtestijn RSP. (Ed. allometry. 147–157. 1129–1136. Functional Ecology 7. Garnier E. M Franco) pp.). Ryser P (2000) Root tissue structure is linked to ecological strategies Thompson K. MMI Van Vuuren) Taiz L.1111/j. Global Change Biology 5.2012. doi:10. Díaz S.New handbook for measurement of plant traits Australian Journal of Botany 231 Sperry JS. Palaeoecology 149.1365-2486. Hacke UG (2008b) Evaluation of centrifugal Tyree MT.2006. Mosbrugger V (1999) Leaf venation density as a climate and Srivastava DS. CA) Veneklaas EJ. 723–741.1046/j. doi:10.1111/j. C4 and C3 grasses (Poaceae) species to elevated atmospheric CO2 doi:10. cnr. Cornwell WK. persistence in the soil.1007/BF00296704 concentration: a meta-analytical test of current theories and Turner IM (1994) Sclerophylly: primarily protective? Functional Ecology 8. M Ciamporová. 191–205. Klimeš L.

125–159. Science 291. doi:10. Physiologia Plantarum 127.2307/1932049 doi:10. (Macmillan Veneklaas EJ.1086/303346 Wright IJ. 534–543. Wong S-C.1996. 1003–1015. 403–416. Cunningham SA.1057175 351–359. 649–656. doi:10.1016/S1360-1385(00)01838-0 during ontogeny. Reich PB (2000) The time value of leaf area.92. doi:10. 486–493. Tjoelker MG. Westoby M. Navas ML. Lamont BB (1991) Leaf specific mass confounds leaf density correlates of early seedling morphology in east African trees and shrubs.2001. Functional Ecology 15. Muller-Landau HC. 213–227. Holbrook NM (2002) Wright W.2002. doi:10.1111/j. Zimmermann MH. doi:10. Thomas SC. Oikos plants. and ambient partial pressure of CO2 doi:10. Vriesendorp CF.1104/pp. Influence of nitrogen nutrition. 972–978. Annals of Botany 99. doi:10.ecolsys. Clifford HT. Keddy PA (1998) Community assembly rules. Kidson R. photon flux density. Reich B.33. Westoby M. Cowan IR.6. Yeo AR (1983) Salinity resistance: physiologies and prices.tb01280. American Journal of Botany 92. Reich PB (2002) Convergence towards higher leaf plants’. Influences of water stress and doi:10. 821–827.1095-8312. Kitajima K (2005) Evolutionary and ecological Witkowski ETF. Harms KE.2007.1111/j. . American Journal of Botany 97. Plant. Poorter doi:10.2307/2390573 Environment 25.tb04172. Annual Garnier E. 381–397.1890/0012-9615(2006)076[0381:COSALS]2. and the coexistence of plant species.Correctly.2307/3237076 Wright IJ. Ackerly DD. Westoby M. doi:10. Journal of Ecology 90. doi:10. Westoby M (2002) Leaves at low versus high rainfall: coordination (1999) Challenging Theophrastus: a common core list of plant traits for of structure. Wright IJ. Eriksson O Wright IJ.1139/b81-248 rate of CO2 assimilation.821 some American woody plants.1093/aob/mcl066 structure and life span in roots and leaves among temperate trees. Wright W. Withington JM.3732/ajb. 539–566 mass per area in dry and nutrient-poor habitats has different consequences (Backhuys Publishers: Leiden. Overton J. Thompson K. Lee W.1126/science. 1011–1022.x plant hydraulics. Weiher E. Melcher PJ.1999. Journal of Ecology 96. Webb CO. Functional Ecology 9.1365-2745.x doi:10. Arnold SEJ. doi:10. Villar R (2004) The worldwide leaf economics spectrum. Boyce CK. 424–426.1104/pp. lifespan and physiology.1111/j. Pyankov VI. Linnean Phylogeny and variation in light capture area deployed per unit Society of London 69. In ‘Variation in growth rate and productivity of higher Wright IJ. Moles A. 830–834. Oleksyn J. Diemer M. Flexas J.x doi:10.1038/nature02403 Will H.3732/ajb. and thickness.tb01222. Chapin T. Mazer SJ.00699. Falster D. Sack L.2002.1469-8137.1046/j. Poorter L. 1059–1062. Farquhar GD (1985c) Leaf conductance in relation vascular epiphytes from central Panama. Williamson GB. Ziegler H (1997) The occurrence of crassulacean metabolism among Wong S-C. investment in leaves: designs for selecting study species with a view to doi:10.x .00427. the coordination between leaf and stem traits. Falster DS. Gulias J. Poot P.4. Westoby M. Reich PB. with photosynthetic capacity. 445–456. Ackerly D. doi:10. Groom PK.x Zwieniecki MA. Eissenstat DM (2006) Comparisons of doi:10. Wilson PJ. Wright SJ (2007) matter content as alternative predictors of plant strategies. Pickup M. Ibarra-Manríquez G. Ecology 43. doi:10. functional ecology. Roderick M.150452 H. Lamont BB.232 Australian Journal of Botany N. Farquhar GD (1985a) Leaf conductance in relation American Journal of Botany 97. doi:10.1983. doi:10.1046/j.x generalizing. Melcher PJ. doi:10.x American Naturalist 155. Vesk P. Cowan IR.x Weiher E. Chapman CA. Westoby M. New Phytologist 137.3732/ajb. Oecologia 88.’ 2nd edn. strategies: some leading dimensions of variation between species. doi:10.x photoinhibition. . Wright I (2002) Plant ecological Cavender-Bares J. I.1046/j. Plant Physiology 78. properties. Jeje AA (1981) Vessel-length distribution in stems of doi:10. Farquhar GD (1979) Stomatal conductance correlates Zanne AE. Martínez- doi:10. Lusk C.1038/282424a0 anatomy and their relation to wood density and potential conductivity.2000. Physiologia Ecological Monographs 76. Baruch Z.2307/3547051 401–413. Roumet C.826 Zotz G. sclerophyll flora. 519–524. Midgley JJ. Trends in Plant Science 6. 826–829. Thompson K.972 Wong S-C. 130–133. Wright IJ (1998) evolutionary lineages. Farquhar GD (1985b) Leaf conductance in relation to 1882–1892.4.1469-185X.1046/j. 66–71. doi:10.1365-3040. Canadian Journal of Botany 59.1399-3054. Clarke GDP. 309–322.1111/j.2 Zanne AE.0900243 Ramos M.00479. MMI Van Vuuren) pp. New Phytologist Relationships among ecologically important dimensions of plant trait 143. Fonseca C. 1445–1450. Oleksyn J.x gravity. Cell & 5 grasses. Reed ML. Pitman NCA. 155–162. Cannon K (2001) Relationships between leaf lifespan and structural hydraulic resistance in plants. 207–215. Biological Journal of the Linnean Society. Warton D. 609–620.x variation in seven neotropical forests. 821–825. Falster DS. Holbrook NM (2001) Hydrogel control of xylem Wright IJ.01341. doi:10. Plant Physiology 78. Publishing: New York) Nature 428. Cornelissen JHC.1399-3054.00689. H Poorter H. Hikosaka K.010802. Garnier E. A survey of seed and seedling characters in 1744 Australian dicotyledon Plant and Soil 199. Plant Physiology 78. Coomes DA (2010) Angiosperm wood structure: global patterns in vessel doi:10.1146/annurev. 214–222.830 Zwieniecki MA. Westoby M (2000) Westoby M (1998) A leaf-height-seed (LHS) plant ecology strategy scheme. nutrition. Plantarum Journal of Vegetation Science 10.CO.1046/j.1365-2435. Loarie SR.0900178 to rate of CO2 assimilation.00922. Cowan IR. Effects of short-term exposures to different Zinke PJ (1962) The pattern of influence of individual forest trees on soil photon flux densities. II. Wong S-C.00522. Nature 282. Prior L. and their implications for doi:10. Illius AW (1995) A comparative study of the fracture properties of Hydraulic architecture of leaf venation in Laurus nobilis L. to rate of CO2 assimilation.2006. 223–229.1469-8137. defences in a low-nutrient. Bongers F.2002.00800. 81. Ackerly DD. Hodgson JG (1999) Specific leaf area and leaf dry Silman MR. Vincent JFK (1996) Herbivory and the mechanics of fracture in morphological dispersion. Westoby M. III. Wiemann MC (2010) Measuring wood specific doi:10.1365-2745. Biological Reviews of the Cambridge Philosophical Society 71.78. Whittaker RH (1975) ‘Communities and ecosystems. Van der Werf A.1046/j. Pérez-Harguindeguy et al.4. Bongers F. Osada N. Lee T. Cowan IR. Paz H. doi:10.1023/A:1004327224729 species: cross-species trait correlations and correlated trait-shifts within Westoby M.78. New Phytologist 155.1111/j. Niinemets U.1469-8137. Review of Ecology and Systematics 33. (Eds H Lambers. phosphorus Zhu JK (2001) Plant salt tolerance. Rice BL. 521–547. The Netherlands) for leaf life span. Westoby M (2006) Cross-species patterns in dispersal by animals.1997. Tackenberg O (2008) A mechanistic simulation model of seed Wright IJ. 269–278.

8 Leaf to sapwood area Unitless 100–103 5 10 – 2. 5 10.d.03–1.2 Life form Categorical – 3 5 – 2.7 Physical strength of leaves 5.5 Clonality Categorical – 5 10 – 2. 5 10.01–140 10 25 17–36 2.1–5B 5 10 3. it corresponds to the number of individuals (=replicates). 5 10.15–0.40 5 10 – (for seedlings. Numbering of plant traits corresponds with the numbering of the chapters in the handbook Plant trait Preferred unit Range of values Recommended no. Note that one replicate can be compounded from several individuals (for smaller species).5–6. 40 10.10 Vein density mm mm–2 0. down to 0. Summary of plant traits Summary of plant traits included in the handbook The range of values corresponds to those generally reported for field-grown plants. of ramifications 0 – >100 5 10 – per branch 2. 4 10–28 3. when two values are given.5–30 Spine length : leaf length Unitless 0–30 2.5 Force to punch N mm–1 0.17–40 Work to shear J m–1 0. The expected coefficient of variation (CV) range gives the 20th and the 80th percentile of the CV (=s.13 Water-flux traits 10 20 Gap fraction Unitless 0–1 Stem flow % 0–50 Water retention on plant surface g m–2 0–500 Leaf wettability degrees (contact angle) 0–180 Droplet retention ability degrees (slope angle) 0–90 3 Leaf traits 3.6 3. 160 11–39 (b) Duration of green foliage Month 1–12 5 10 – 3. scaled to the mean) as observed in a number of datasets obtained for a range of field plants for different biomes.2 Area of a leaf mm2 1 – >206 5.2–5 5.7 Branching architecture No. 4 14–29 Force to tear N mm1 0.9 Photosynthetic pathway Categorical – 3 3 – 3.1 Specific leaf area m2 kg1 (mm2 mg1)A <1–300 5. whereas one individual cannot be used for different replicates.10 Salt-tolerance traits 5 10 Selective root cation uptake Unitless – – Salt excretion and compartmentalisation Categorical – – 2.New handbook for measurement of plant traits Australian Journal of Botany 233 Appendix 1. 5 10.02–0. 4 8–16 3.5 pH of green leaves or leaf litter Unitless 3.5–200 5.4 Plant height m <0.4 Leaf thickness mm <0. 4 17–36 3. 4 1–6 3.13 Electrolyte leakage % 2–100 5.3 Leaf dry-matter content mg g1 50–700 5.4–4 5 10 3.5 5. 5 10.11 Relative growth rate mg g–1 day–1 2–300 10 20 2.11 Light-saturated photosynthetic rate mmol m–2 s–1 2–30 5 10 3. 5 10.1 Life history Categorical – 3 5 – 2.10) 2. Recommended sample size indicates the minimum and preferred number of individuals to be sampled.6 Spinescence 5 10 – Spine length mm 0.6 Leaf nitrogen and phosphorus concentrations (a) Leaf nitrogen concentration mg g1 5–70 5. 5 10.5–25 5 10 3.12 Shoot flammability Unitless 0 – ~3 5 10 2.9 Root-mass fraction Unitless 0. CV of replicates range Minimum Preferred (%) 2 Whole-plant traits 2. when only one value is given. 4 9–26 (Continued ) . so as to obtain an appropriate indication of the values for the trait of interest.5–300 Spine width mm 0.3 Growth form Categorical – 3 5 – 2. Ranges of values are based on the literature and the authors’ datasets and do not always necessarily correspond to the widest ranges that exist in nature or are theoretically possible. 4 4–10 3. 5 10.8 Leaf lifespan and duration of green foliage (a) Leaf lifespan Month 0. the first one corresponds to the number of individuals and the second one to the number of organs to be measured per individual. 4 8–19 (b) Leaf phosphorus concentration mg g1 0.12 Leaf dark respiration mmol m–2 s–1 0.

5 Seedling morphology Categorical – 3 6 – 6. Pérez-Harguindeguy et al.16 Litter decomposabilityC %Mass loss 0–100 10 20 7–14 4 Stem traits 4.2 Root-system morphology 5 10 Depth m 0.6 Resprouting capacity Unitless 0–100 5 10 – A Alternative preferred units in parentheses. 10 11–33 3.4 Xylem conductivity 5 10 21–63 Stem-specific xylem hydraulic kg m–1 s–1 MPa–1 1 (gymnosperms) conductivity (KS) to 200 (tropical lianas) Leaf-area-specific xylem hydraulic kg m–1 s–1 MPa–1 6  10–5 conductivity (KL) (gymnosperms) to 1  10–2 (tropical lianas) 4.3 Bark thickness mm .3 Dispersal potential Dispersules – 10 20 – dispersed/dispersules produced 6.1 Dispersal mode Categorical – 3 6 – 6.csiro.234 Australian Journal of Botany N. CV of replicates range Minimum Preferred (%) 3. number of leaves in each sample will depend on its weight and the size of the litterbag. B Considering only photosynthetic tissue. (continued ) Plant trait Preferred unit Range of values Recommended no.05–70 Lateral extent m 0.05–40 Density of exploration mm mm–3 10–4–1 5.1–1. 10 15–24 5.3 Nutrient uptake strategy Categorical – 5 10 – 6 Regenerative traits 6.25–14 5 10 20–45 5 Below-ground traits 5.3 5 10 5–9 4.publish.1 – >30 5 10 4. www. C Replicate numbers correspond to the number of individual plants (replicates) from which to collect leaf litter. Appendix 1. 5 5.2 Dispersule size and shape Size (mass) mg (g)A 5 10 Shape Unitless 0–1 3 6 – 6.2 Twig dry-matter content mg g1 150–850 5 10 2–8 4.15 Leaf palatability %Leaf area consumed 0–100 10 20 3.5 Vulnerability to embolism (Y50) MPa –0. 10 10.4 Seed mass mg 103–107 5 10 14–27 6.14 Leaf water potential MPa 7!0 5. total leaf thickness can be >40 mm in some succulent plants.1 Stem-specific density mg mm3 (kg l1)A 0.1 Specific root length m g–1 3–350 5.