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Journal of Biological Education

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Teaching botany on the sunny side

of the tree: promoting investigative
studies of plant ecophysiology through
observations and experiments on sun
and shade leaves
a b
Jonathan D. B. Weyers , Hans-Olof Höglund & Birgitta McEwen
b

a
Department of Biological Sciences , University of Dundee ,
Dundee, Scotland , DD1 4HN
b
Department of Natural Sciences , University of Karlstad , S-651
88 Karlstad, Sweden
Published online: 13 Dec 2010.

To cite this article: Jonathan D. B. Weyers , Hans-Olof Höglund & Birgitta McEwen (1998) Teaching
botany on the sunny side of the tree: promoting investigative studies of plant ecophysiology
through observations and experiments on sun and shade leaves, Journal of Biological Education,
32:3, 181-190, DOI: 10.1080/00219266.1998.9655619

To link to this article: http://dx.doi.org/10.1080/00219266.1998.9655619

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 Teaching botany on the
sunny side of the tree:
promoting investigative
studies of plant
Downloaded by [b-on: Biblioteca do conhecimento online UA] at 02:21 05 December 2014

ecophysiology through
observations and
experiments on sun and
shade leaves
Jonathan D. B. Weyers, Hans-Olof Hoglund, and Birgitta McEwen

Investigations on sun and shade leaves allow students to compare plant

growth patterns in different environments and to appreciate plants as
slow-motion combatants for light as a resource

Abstract
Plants can respond developmental^ to many
environmental stimuli. Sun and shade leaves are an
example of an acclimation to light intensity. This paper
describes how this phenomenon can be used as a
stimulating vehicle for teaching plant ecophysiology to
undergraduates. Detailed protocols for investigations are
outlined, with an appendix relating to studies in schools.
We also provide representative data and introduce topics
for classroom or tutorial discussion.
Key words: Leaf development, Light environment, Plant
acclimation.
Abbreviations: A: net assimilation rate; ASV: air space
volume; C,: internal C 0 2 concentration; DM: dry mass;
FM: fresh mass; a,: leaf water vapour conductance; HI;
harvest index; IM: infiltrated mass; IRGA: infra-red gas
analyser; LA: leaf area; LAI: leaf area index; PAR:
photosynthetically active radiation; PI: plasticity index;
PPFD: photosynthetically active photon flux density; RSR:
root-to-shoot ratio; RTR: root-to-total-mass ratio; SLA:
specific leaf area; SRR: shoot-to-root ratio.
Introduction
For many years it has been known not only that there
are 'sun' and 'shade' species of plant — those that are
better adapted to high or low light environments —
but also that individual plants can respond ('accli­
mate') to high and low light regimes by producing
specialized sun and shade leaves. Indeed, the compar­
ison between transverse sections of sun and shade
leaves (e.g. in beech, Fagus sylvatica, figure 1) has
become a classic topic in plant anatomy. It is, how­
ever, possible to take the anatomical and physiologi­
cal comparison between these leaf forms much further
than the drawing of simple tissue diagrams. Sun and
shade leaves exhibit fascinating differences in a range
of characters that can be related, both in the course of
investigative learning and during the ensuing class­
room or tutorial discussions, to the leaf and plant
carbon budget, the effects of one plant on the growth
of others nearby, and ultimately to the evolutionary
success of the organism. Furthermore, such studies
provide an interesting and meaningful context for
teaching a wide range of practical skills.

Journal of Biological Education (1998) 32 (3) 181

 Teaching botany on the sunny side of the tree
SUN LEAF
100
Hm
Intercellular air spaces
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Figure 1 Transverse sections of sun and shade leaves of beech, Fagus sylvatica L. Line drawing based on a photomicrograph of
a slide obtained from Philip Harris Educational. Lichfield, UK (Cat. No. H42725/6), showing typical anatomical differences between
the leaf morphs. Note that the decreased thickness of the shade leaf (ca. 73 per cent) is due mainly to changes in the thickness of pal­
isade mesophyll, which also consists of one layer of cells instead of two.
The aim of this article is to suggest possible ap­
proaches for using the topic of sun and shade leaves as
a practical or project exercise, based on our experi­
ences during a pilot undergraduate course at the
University of Karlstad, Sweden. Topics that can be
studied include in approximate order of sophistication:
• determination of standard measures of growth such
as leaf area and thickness;
• measurement of more complex indices of growth
relevant to ecosystem or crop performance;
• use of a radiometer to estimate the absorbance of
whole leaves and measure photosynthetically
active light (PAR) at different spots in the plant
canopy;
• design of appropriate sampling procedures or field
experiments to take into account heterogeneity in
the growth environment;
• statistical analysis of data by Student's Mest;
• quantitative studies on chlorophyll content; and
• advanced measurements of photosynthetic charac­
teristics.
Studies on sun and shade leaves can be made in the
field on local deciduous trees if the season permits.
Alternatively, they can be based on simulated field
experiments involving the growth of a crop species
under different light regimes.
Materials and methods used to obtain
illustrative results
Plant material
Radish plants (Raphanus sativus cv. 'Pernod', Svalof
Weibull AB, Sweden) were raised from seed at the
University of Karlstad in July 1996. Five seeds were
planted per 100 mm plastic pot in a peat-based com­
182 Journal of Biological Education (1998) 32 (3)
Weyers, Hoglund, and McEwen
SHADE LEAF
Upper epidermis
Palisade mesophyll
Spongy mesophyll -
Lower epidermis —
post. They were allowed to germinate in full sunlight
in a south-west facing glasshouse. Water was pro­
vided by capillary matting. The glasshouse was well
ventilated, but no supplementary light was provided.
After three weeks, the seedlings were thinned to three
per pot and thereafter were fed twice-weekly with 2.5
strength 'Superbe' 8:4:4 N:P:K feed (Svalof Weibull
AB, Sweden; similar to 'Liquid Gromore' and
'Fison's Liquinure'). At this time, the pots were ran­
domly assigned to sun or shade conditions. The 'sun'
plants were grown on the glasshouse bench as before,
fully exposed to ambient sunlight. The shade plants
were grown beneath the same bench in a tent consist­
ing of four layers of white garden fleece material, and
one layer of green wind break material. This reduced
the photosynthetically active photon flux density
(PPFD) to 8-11 per cent of that in the sun environ­
ment. Plants were used in experiments 2-5 weeks fol­
lowing assignments to treatments.
Specimens of various plants were selected for study
on or near to the campus of the University of Karlstad
(latitude 59° 24' N, longitude 13° 35' E). They were
identified using standard Swedish floras. Criteria for
selection of specimens included:
• density of crown, ideally providing dense shade to
contrast with exposed sun positions;
• existence of sun and shade leaf forms as judged
subjectively through appearance and feel;
• accessibility of the specimen for on-site studies; and
• the species' leaf morphology, including size, shape,
and hairiness.
Studies on leaf morphology
The mass of leaves and other plant parts was deter­
mined on a self-tareing milligram or tenth milligram
 Teaching botany on the sunny side of the tree Weyers, Hoglund, and McEwen
balance. For fresh mass (FM) determinations, speci­
mens were weighed as soon as possible after detach­
ing, those from the field being enclosed in polythene
bags between harvest and weighing. Leaf area (LA)
was then estimated by photocopying, weighing cut­
outs, and comparing these to the mass of a standard
area of paper. Dry mass (DM) was determined after
oven-drying at 80 CC for at least 24 h. In separate sam­
ples, the percentage air space volume (%ASV) of
leaves was estimated from the difference between the
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infiltrated mass (IM) of the leaf (or leaf discs, see
below) and the FM. The IM was found by weighing
after alternating exposure to low pressure and atmos­
pheric pressure, while the leaf was under water in a
side-arm flask. The total volume of the leaf or disc
was derived from the IM assuming a mean density for
infiltrated leaf material of 1000 kg m"3. Leaf thickness
was determined from hand sections while viewing
under light microscopy with a calibrated eyepiece
graticule.
Leaf absorbance and chlorophyll determinations
Simplified measurements of leaf absorbance were
obtained by taking a radiometer reading with and
without a leaf covering the sensor. To assay chloro­
phyll, four or five discs were taken per leaf using a
15 mm diameter cork borer, immediately weighed
and then placed in ca. 20 ml 95 per cent ethanol. This
was then boiled to extract the pigments. Any solvent
that evaporated was replaced from time to time to
avoid drying down. A fume cupboard was used to
avoid exposure to solvents. When the discs appeared
devoid of green pigment, the specimen was made up
to a known volume and its absorbance measured at
645, 649, and 665 nm. Leaves from some tree
species proved recalcitrant compared to radish and
required up to one hour extraction. As chlorophyll
degrades in light, all samples were protected by
wrapping flasks in aluminium foil throughout these
operations. Chlorophyll concentrations in the spec-
trophotometer cuvettes were estimated from the fol­
lowing formulae according to Wintermans and de
Mots (1965): chlorophyll a (mg 1"') = 1 3 ^ ^ -
5.76AM9; chlorophyll b (mg l"1) = 25-SA^ - 7.6A665.
The formula mass of chlorophyll was taken as 894 g
mol"1. Note: it is also possible to extract chlorophyll
in 80 per cent acetone, but different formulae should
be used.
Leaf gas exchange
Gas exchange under controlled conditions was
measured using a portable CIRAS-1 infra-red gas
analysis system fitted with a broad leaf clamp (area
2.50 cm2) and standard light unit (PP Systems,
Hitchin, UK). To estimate photosynthetic characteris­
tics of the leaves, two types of experiment were car­
ried out: firstly, an 'AJg,' curve was constructed by
subjecting the enclosed leaf area to a PPFD of 1022
umol photons m"2 s~' at an RH of 70 or 80 per cent
and a pC0 2 of 355 umol mol"1 (Parsons et al, 1997).
The plot of assimilation (A) against leaf conductance
(gj) was used to estimate the threshold of stomatal
limitation to photosynthesis, and the subsequent A to
PPFD determination was always carried out at a g,
where the stomatal limitation would have been at
most 10 per cent. An 'A/PPFD' curve (also termed
an 'A/Q' curve by some authors) was constructed by
altering the incident PPFD using the manufacturer's
neutral density filters, care being taken not to expose
leaves to high PPFD values for long. From this
curve, values for photosynthetic parameters were
found as follows:
• PPFD saturated rate of net photosynthesis — the
upper asymptote of the relevant curve (generally
the value found at full sunlight);
• PPFD compensation point - intercept with the x-
axis where A = 0;
• photosynthetic efficiency on a per quantum basis —
gradient of curve at the compensation point (see
Parsons et al, 1997).
Analysis of data
Derived indices of growth were calculated as follows
(Hunt, 1990):
Specific leaf area (SLA) = LA/DM (expressed in
m2 g"');
Leaf area index (LAI) = Total LA for plant/surface
area occupied by plant (dimensionless);
Harvest index (HI) = ratio of FM harvestable product
(in this case the 'radish' portion) to total FM
(dimensionless);
Shoot-to-root ratio (SRR) = ratio of root DM to shoot
DM (dimensionless) - in this instance the 'radish'
portion, as thickened hypocotyl, was designated as
shoot; and
Plasticity index (PI) for any measured or derived pair
of sun and shade characters = (higher value - lower
value)/higher value (dimensionless).
Statistical comparisons between mean data for sun
and shade leaves were carried out using the 'TTEST'
function on Excel 5.0 (Microsoft Corp., Redmond,
USA).
Relevant data that can be obtained by
students
Leaf characteristics
The most obvious morphological alteration to leaves
when they develop in shade is that leaf thickness is
reduced. This occurs due to a reduction in the thick­
ness and degree of differentiation in the mesophyll
layers, which is visible in transverse sections (figure
1). Two other ways in which this can be shown are
the specific leaf area, which demonstrates the leaf
area obtained by the plant for a given dry matter
Journal of Biological Education (19981 32 (3) 183
 Teaching botany on the sunny side of the tree Weyers, Hoglund, and McEwen

'investment' and the chlorophyll content per unit leaf
area (see below). Both of these measures are gener­
ally lower in shade leaves and they indicate that the
plant has produced these leaves 'on the cheap' com­
pared to sun leaves. In some species, there can be an
increase in the percentage air space volume
(%ASV), which may also be symptomatic of a
change in mesophyll differentiation. Often, the shade
condition is characterised by an increase in mean leaf
area, though this is not always the case. If leaves are
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experiment like the one on radish (table 1) we
expected that the reduced growth of the shaded plants
will be reflected in a lower DM for the plant for the
simple reason that plants need an adequate supply of
photons to turn C 0 2 and H 2 0 and nutrients into carbo­
hydrates, proteins, etc. The leaf area index (LAI) can
be used to highlight the difference in leaf growth in
the two cases. A LAI greater than one indicates that
there are multiple layers of leaves over any one piece
of ground (mature crops often have values in excess of

larger, of course, they are capable of intercepting
more light, and if thinner and lighter, they can maxi­
mize their use of the available light and not waste
resources on trying to absorb light that is not there.
Measures of whole leaf dry mass and whole leaf
chlorophyll may indicate that the available 'leaf unit'
of dry matter or pigment has simply been spread over
a larger area (cf. specific leaf area). If not, then there
may have been some compensation for the shade
condition.
Partitioning of assimilates
When considering the growth of whole plants and rel­
ative partitioning of resources within the plant body,
the total dry mass of the plant, and the root-to-shoot
ratio or the root-to-total-mass ratio (RSR and RTR
respectively) will provide useful information. In an
four — see Larcher, 1995). This concept can be used
to drive home the message that even in one plant there
is more than one light environment. However, despite
the much lower leaf area and leaf DM in the shade
plants, it may be possible to show that the SRR is
higher in the shaded plants. They appear to have com­
promised some root growth in favour of organs that
will capture the scarce (limiting) resource of light. It
should be noted that SRR measurements are suscepti­
ble to error in determination of root DM.
With a quickly maturing plant like radish it is also
possible to illustrate the profound effects of light on
the harvest index, where the harvestable 'product' is
the radish itself detached from leaves and roots. This
component is practically negligible for the shade plants
in the experiment shown in table 1, again reflecting the
shaded plants' prioritization of resources.

Table 1 Growth indices for plants of radish, Raphanus sativus L. subjected to 'sun' and 'shade' environments. Values are means of six replicates
per light regime except where indicated in parentheses. For results of Student's /-test: NS = P>0.05 (not significant); * = P <0.05; ** = P <0.01;
*** = p <0.001; — = not relevant (qualitative data)

Light regime

Growth index (units) Sun Shade Plast

Plasticity index Result of Student's Mest
1
Part 1 Fresh mass (gFM plant" ) 51.3 11.1 0.78
1
Dry mass (gDM plant" ) 3.83 0.59 0.85
2 1
Leaf area (cm plant" ) 513 220 0.57

Leaf thickness (mm) 0.34 0.25 0.27

Number of palisade mesophyll layers; degree

of differentiation 2-3; well differentiated 0-1; poorly differentiated —

Leaf air space volume (%) 27.4 24.6 0.10 NS

2 1
Part 2 Specific leaf area (m kg DM" ) 14.2 37.5 0.62

Leaf area index 3.2 1.4 0.57

Shoot-to-root DM ratio 34.4 18.6 0.46

Root-DM to total DM ratio 0.03 0.06 0.51 NS

Harvest index on FM basis 0.46 0.06 0.87

Leaf absorbance (%) 87 81 0.07 ** («=5)

2
Chlorophyll (a+b) content (mmol m~ ) 0.21 0.16 0.26

Chlorophyll a:b ratio 6.5 5.4 0.17

184 Journal of Biological Education (1998) 32 (3)

 Teaching botany on the sunny side of the tree Weyers, Hoglund, and McEwen

Pigments and photosynthesis proportion of chlorophyll b than do photosystems I

Absorbance and pigment measurements provide
opportunities to explore the responses of the plant in
terms of light-capturing efficiency. The absorbance of
the leaf gives a simple indication of its pigment con­
centration on an area basis. A more sophisticated way
of showing this is via measurements of chlorophyll
(a+b). The more advanced students will be able to
grasp this as a cost-benefit problem; a thicker, pig­
ment-rich leaf would be more efficient at absorbing
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and II, a decrease in the a:b ratio indicates a propor­
tionally higher number of light harvesting complexes
than photosystem units (see Lawlor, 1993).
Gas exchange measurements require sophisticated
equipment and may also require an advanced know­
ledge base on the part of students to understand their
full meaning. If students cannot collect their own data,
then discussing model curves like those shown in
figure 2 might suffice. We have restricted this presen­

light, but it might cost too much in resources to cap­
ture the last few per cent of the incident radiation.
Further, these additional 'layers' of a sun leaf might
not actually make a significant net contribution to the
carbon budget, because of innately high respiration
rates (see below), We can assume that Natural
Selection has acted to ensure that the most efficient
'design' survives. The chlorophyll a:b ratio requires
discussion at a rather deeper level and a full under­
standing will be dependent on background knowledge
concerning the pigment-protein complexes in the
granal and stromal thylakoids in chloropolasts.
Because light-harvesting complexes contain a higher
tation to the A/PPFD curves for sun and shade leaves,
since these demonstrate most of the important facts of
the sun and shade syndromes. The A/C; curve is simi­
lar in general form (see Lawlor, 1993), with the differ­
ences in derived parameters between the two curves
being for essentially similar reasons.
The A/PPFD curves are constructed under natural
atmospheric concentrations of C0 2 so the photosyn-
thetic capacity evident in the upper asymptote repre­
sents the maximum rate of photosynthesis attainable
in vivo (assuming other conditions such as water sta­
tus and boundary layer thickness are not limiting).
The differences between the two leaf forms can be

Asymptote = PPFD-saturated rate of photosynthesis

TYPICAL SUN LEAF CURVE

u
e TYPICAL SHADE LEAF CURVE
>>
Q
O

■a Slope = photosynthetic efficiency on a per quantum basis

o
a

Zero intercept = PPFD compensation point

400 800 1200 1600 2000

Photosynthetically active photon flux density
Figure 2 Model 'A/PPFD' curves for light-dependent photosynthesis in sun and shade leaves.
Typical curves are shown for both leaf morphs. As illustrated on the sun leaf curve, three important photo-
synthetic characteristics that can be derived from such data are: (a) the PPFD-saturated rate of net photo­
synthesis (units: mol C0 2 m~2 s"'), which is the asymptote of the curve; (b) the PPFD compensation point
(units: mol photons m 2 s~'), which is the point at which net photosynthesis is zero; and (c) the photosyn­
thetic efficiency on a quantum basis (units: mol C0 2 (mol photons)"1), which is the slope of the curve at the
light compensation point. Note that for certain PPFD values at the low end of the natural range, the shade
leaf will out-perform the sun leaf. One of the reasons for this is evident from a comparison of the expected
rates of net photosynthesis in the dark — this indicates that the respiration rate of the shade leaf is lower on
an area basis than that of the sun leaf.

Journal of Biological Education (1998) 32 (31 185

 Teaching botany on the sunny side of the tree
attributed to their differences in pigment and dry mat­
ter as previously discussed — there is simply less pho-
tosynthetic machinery available on an area basis in a
shade leaf. The smarter students may see immediately
that light saturates photosynthesis in both cases well
below full sunlight (the shade leaf generally being sat­
urated at a lower PPFD than the sun leaf), and can
then be asked what they think limits photosynthesis
above this value. The answer can be deduced easily
from an A/C; curve (see Lawlor, 1993), for the upper
asymptote in this case represents the potential photo-
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synthetic capacity when C0 2 is not limiting (i.e. well
above atmospheric concentrations) and is far higher
than the PPFD-saturated value (in C3 plants in the pre­
sent atmosphere). This can lead on to interesting spec­
ulation on the possible effects of the rise in pC0 2 due
to the use of fossil fuels.
The difference in PPFD compensation point
between sun and shade leaves is vital to understanding
the role of the latter in the plant's carbon economy.
The 'lean' nature of shade leaves means that they
respire less both on area and DM bases. Their break­
even requirement for light is therefore much lower
than that for sun plants. Inspection of the two A/PPFD
curves (figure 2) reveals that there is a range of PPFD
values where the shade leaf actually out-performs the
sun leaf, and a further range where their rates of pho­
tosynthesis are comparable. These PPFD ranges cor­
respond to shade conditions (i.e. that produced by
approximately one 'layer' of leaf above). Now it can
be seen that far from being a potential liability, the
shade leaf syndrome is essential if the plant is to ben­
efit (at least in carbon budget terms) from having a
multi-layered canopy.
There may or may not be differences in the photo-
synthetic efficiency on a quantum basis (note: ideally
these values should be obtained from a plot of absorbed
PPFD rather than incident). Researchers have not
found any clear evidence to date that indicates a funda­
mental difference between the two leaf forms.
Numerical analysis
The notion of putting numbers to observations is a key
theme of modern plant ecophysiology and it is worth
emphasizing to students that a quantitative approach
is required in most aspects of biology. The need to
analyse inherently variable results via statistics is also
fundamental. One useful outcome of the binary nature
of the sun/shade condition (albeit contrived because it
ignores intermediate conditions and the temporal and
spatial variation in the light environment) is that com­
parisons between data for the two conditions can be
made using Student's f-test or an equivalent non-para­
metric method. In using such tests on their own data,
students come to appreciate, in a manner that does not
involve overly complex mathematics, how the deci­
sion about whether two samples differ is a matter of
probability.
186 Journal of Biological Education (1998) 32 (3)
Weyers, Hoglund, and McEwen
Which of the above indices shows the greatest sus­
ceptibility to shade? Or, comparing species studied by
a group of students, which shows the greatest
response to shade for a selected index? Both of these
questions can be answered by calculating the plastic­
ity index in each case. This index is used to 'normal­
ize' the comparisons across different units. If shade
has no effect on the measured character, then the PI
will be zero; and the greater the effect, the closer the
PI approaches one. For example, in the radish experi­
ment described in table 1, the shade treatment caused
a greater relative effect on the dry mass (PI = 0.85)
compared with the fresh mass (PI = 0.78). Note that
the PI is not always indicative of the degree of statis­
tical significance, since the latter depends not only on
the difference between mean values but also the vari­
ation in the samples (compare results for SSR and
RTR in table 1).
Suggested protocols for investigations,
witn representative results
Novice undergraduates
These students should be able to carry out all of the
above work onfield-derivedspecimens. They should
be able to calculate the more complex growth indices,
to consider what they will tell them about the nature of
leaf or plant growth and to determine the relative
effects of shading using the plasticity index. If a quan­
tum sensor (preferably measuring PAR) is available,
the consequences of these pigment differences can be
seen in the absorbance of PPFD by whole leaves.
Students at this stage should be capable of collecting
all data in table 1 and able to write up their work as an
assessed report. They should be able to discuss their
results in an ecophysiological context at some depth.
If a spectrophotometer is available, pigment concen­
trations can be measured, and the concepts of
absorbance, absorbance spectra, and Beer's law can
be seen in action. Less sophisticated measurements of
pigment content might be carried out using a col­
orimeter or similar instrument (it will not then be pos­
sible to differentiate between chlorophyll a and b).
Students at this level should be capable of appreci­
ating the problems involved in sampling from nature.
Measurements of PAR, for instance, provide an
insight not only into variability in the ecosystem but
also into the unreliability of subjective estimates of
light. They should also benefit from involvement in
designing, organizing, maintaining, and deriving data
from a glasshouse (or field) experiment such as the
radish growth experiment summarized in table 1. This
is an opportunity to put into practice 'dry' concepts
like replication, Latin square design, and statistical
analysis. With a real example to aid their imagination,
students can identify more readily with the concepts
of interfering and confounding variables (see Jones
Reed, and Weyers, 1998, for definitions).
 Teaching botany on the sunny side of the tree Weyers, Hoglund, and McEwen

The simulated field experiment can be used to
introduce students to agricultural economics when
working out and discussing the HI and RSR.
However, when obtaining raw data for working out
these parameters, some care is required when separat­
ing the root system from the soil, and use of a well-
fertilized sand-'Perlite' mixture might make this eas­
ier and more accurate than a peat-based compost.
Teachers should not underestimate the time required
to obtain the raw data: the number of replicates used
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is often used in these curves, however, although this is
not too difficult to determine, the incident PPFD
should be used here because results will then better
reflect the in vivo photosynthetic performance of the
leaf forms, including e.g. absorbance and reflectance
properties.
At this stage, the work described above might form
the basis of a research project, providing leaf material
is available during the study period. A major advan­
tage of using this broad subject area is that there is suf­

should be as small as possible, consistent with the
intended statistical analysis.
Advanced undergraduates
In some university departments, a research-grade
instrument like the CIRAS portable IRGA may be
available to students. Their understanding of gas
exchange, the limitations on photosynthesis, and con­
cepts such as stomatal conductance, Q, net and gross
photosynthesis will be advanced dramatically by
using this type of instrument to obtain A/PPFD and
A/Cj curves. If data are graphed as they are obtained,
notions such as compensation point and photosyn-
thetic capacity 'come alive' to students as they manip­
ulate net photosynthetic rate between its minimum
and maximum values. Where such equipment is
accessible, data like those shown in figure 3 can be
obtained. Note: in research work, the absorbed PPFD
ficient variety in potential subject matter to promote
individuality, yet the work of students or groups can
easily be compared using the plasticity index. The
results obtained are suitable for writing up in standard
laboratory reports; at Karlstad, we successfully
assessed such group investigations through the con­
struction and presentation of a poster, involving a
combination of marking from the peer group, the pro­
ject supervisors, and departmental 'visitors' to the
poster session.
Interpretation of results and ideas for
further discussion in classroom or tutorials
Issues concerned with sampling and experimental
design
During field sampling, students can be prompted to
consider why it is important to be able to describe

0 500 1000 1500 2000 2500

Photosynthetically active photon flux density (|amol photons m" s"1) 2

Figure 3 Light-dependent photosynthesis in sun and shade leaves of radish, Raphanus sativus L.
Data obtained by students at the University of Karlstad, Sweden, using the CIRAS-1 IRGA. Sun leaf, open
circles; shade leaf, closed circles. In the shade leaf, the PPFD-saturated rate of net photosynthesis was 43 per
cent of that in the sun leaf (8.7 vs. 20.3 mol C0 2 rrf2 s~'); the PPFD-compensation point was 51 per cent
(20.0 vs. 39.5 mol photons nT s ); and the photosynthetic efficiency was 56 per cent (0.05 vs. 0.09 mol C0 2
(mol photons)-

Journal of Biological Education (1998) 32 (3) 187

 Teaching botany on the sunny side of the tree Weyers, Hoglund, and McEwen

accurately the tree they have visited — not only to
specify its precise location, but also to identify what
species it is, using a suitable flora. Another important
matter to discuss is the ontogeny of the leaves used. It
is easy for students to observe that the character of
leaves differs with insertion point on the axis; older
leaves that are further back are generally both thicker
and greener. They may suggest themselves that 'sys­
tematic' variation like this might be countered by
selecting leaves of equivalent insertion point on sun
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ber of 'confounding' variables also vary with PPFD in
the canopy. The most obvious examples are tempera­
ture and humidity, but there are others. This can lead
neatly to a discussion on the role of experimentation
in biology, where the aim is to control as many of the
important confounding variables as possible to 'pin
down' the important influence. The need for replicate
measurements can be justified relatively easily when
comparing the variation in the growth of plants in the
same environment. Discussion of uncontrolled sys­

and shade branches. This is a useful juncture at which
to point out the fact that a leaf will develop as a sun or
shade leaf according to the light environment it per­
ceives during leaf expansion and differentiation (i.e.
early spring). This acclimation response is a 'once and
for all' irreversible response for any given leaf,
though of course, should conditions change between
years (e.g. the adjacent tree falls down in a winter
gale), the next annual flush of leaves can develop
appropriately.
One benefit of venturing into the field to select
specimens is that the inherent variability in the natural
environment is immediately obvious. Selecting 'sun'
and 'shade' leaves focuses this issue. Discussion can
first centre on what criteria should be used to differen­
tiate these two extreme microclimates (this may differ
among species). Some form of radiometer should be
used to measure PPFD to assist in the decision. A
shade PPFD that is 10 per cent of the direct sky radia­
tion is a reasonable rule of thumb. The 'open sky'
PPFD should be constant between the two measure­
ments — it will be found to change considerably if
cloud cover varies. At this time it may be possible to
observe sunflecks and other dynamic aspects of the
light environment (e.g. time of day, daylength, cloud
cover).
The more astute students may realize that any num­
tematic variation (e.g. in light and temperature) can be
used to introduce notions such as selecting plants for
measurement using a Latin square design, where
every row and column is sampled. Concepts like edge
effects may also be introduced.
Species differences in results
There may be differences in the way in which differ­
ent species respond to sun and shade. It is important
for students not to assume that results they obtain with
a particular species are 'wrong' simply because they
are different from those expected or from those of
their peers. For instance, leaves of many tree species
increase in area under shade conditions (e.g. oak and
birch, table 2), a response that has been interpreted as
an attempt to maximize total photosynthesis per leaf
(as discussed above). However, in other species (e.g.
apple and honeysuckle, table 2), differences may not
be observed. Demonstrating the existence of this
response may in any case be difficult due to natural
differences in leaf area between branches and among
leaves at different stages of development, as well as
variation in the degree of shade. Another example of
response differences concerns effects of shade on pig­
ment contents. Normally, the thinner shade leaves
would be expected to have a lower chlorophyll (a+b)
content on an area basis. This effect was seen in

Table 2 Values of selected growth indices in leaves of four deciduous trees or shrubs growing at or close to the campus of Karlstad University,
Sweden, Values are means of five replicates. PI = plasticity index and P - result of a Student's Mest comparing the mean data for sun and shade leaves,
indicated thus: NS = P>0.05 (not significant); * = P <0.05; ** = P <0.01; *** = P <0.001

Growth index

Plant species Mean leaf area (cm ) Mean leaf thickness (mm) Mean specific leaf area Mean chlorophyll (a+b) content
(m2kgDM"') (mmol nf2)

Sun Shade PI, Sun Shade PI. Sun Shade PI, Sun Shade PI,
P P P P

Oak, 30.9 64.9 0.52, 0.24 0.17 0.31, 9.4 19.4 0.52, 0.50 0.59 0.16,
Quercus robur *t NS
*** ***
Birch, 16.6 23.6 0.30, 0.24 0.16 0.33, 12.3 25.6 0.12, 0.38 0.25 0.25,
Betula pendula ** *** *** **
Apple, 46.1 53.6 0.14, 0.27 0.19 0.29, 12.1 17.9 0.38, 0.75 0.77 0.02,
Malus domestica NS * NS

Honeysuckle, 26.7 27.3 0.02, 0.27 0.17 0.38, 17.0 29.7 0.43, 0.63 0.45 0.29,
Lonicera NS *** *** **
pericylmenum

188 Journal of Biological Education (1998) 32 (3)

 Teaching botany on the sunny side of the tree Weyers, Hoglund, and McEwen
radish, birch, and honeysuckle (tables 1 and 2).
However, in oak and apple, there was no effect of
shade on chlorophyll (a+b) content. One possible rea­
son for this is damage to pigments caused at high
PPFD values.
The individual plant as a competitor for light
If sun leaves absorb about 80-90 per cent of the avail­
able light, why do plants commit resources to capture
some of the remaining 10-20 per cent, rather than, say,
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growing taller or wider, producing more propagules,
or creating larger perennating structures? Of course,
there can be no certain answer to a question like this,
but speculation about plants 'fighting one another' for
the available resources can fire students' imaginations
and encourage them to visualize plants in a more pos­
itive way.
Thefirstthing to be said about having multiple lay­
ers of leaves is that plants in all likelihood make a net
carbon gain from their shade leaves, as explained
above. Research shows that this is not only because
shade leaves are thinner and therefore respire less on
an area basis (seefigure2), but also because they have
lower respiration rates on a gDW basis, implying
some difference in the set-up of the respiratory sys­
tems themselves. The take-home message is that they
require less light to 'break even' on their carbon bud­
get and normally make a net carbon contribution to
the whole plant: just 10 per cent of sunlight is ample
for this.
A second point concerns the nature of the light
environment, which is variable in many ways.
Multiple layers of leaves allow more efficient use of
sunlight as the sun's angle changes during the day.
While most plants arrange their youngest leaves so
that they tend not to overlap (the Fibonacci ratio gov­
erning the angle of insertion), this arrangement is not
perfect. 'Sunflecks' appear in the lower canopy due to
passage of light between leaves and this source of
energy can be 'mopped up' by the shade leaves.
Finally, it is interesting to consider the plant as a
combatant for resources. Students may need prompt­
ing to use their imaginations due to the slower time-
scale involved, but fundamentally the struggle is no
different to that between the Serengeti animals at a
water hole, which are also fighting for a limited
resource (water in that case). The central notion is that
the plant gains in the long run by 'shading out' its
potential competitors. If other plants were able to
grow beneath a tree, they would start to use its other
precious resources such as water and soil nutrients.
The tree's overall growth might suffer as a result (and
hence its ability to pass its genes down to further gen­
erations). Care must be taken in such discussions with
the use of terms such as 'strategy' and 'design'.
Instead, it should be emphasized that we see before us
the results of ruthless selection during millions of
years of evolution. Plants do not 'plan' their responses
in the same way as humans can, but their genotypes
have survived and prospered because of their ability
to grow and reproduce.
Intriguingly, low-growing, under-storey plants
appear to have adapted to this situation. Many pro­
duce seed that responds, via the pigment phy-
tochrome, to the alteration in light quality under a
canopy. As the light is filtered through the leaves, the
chlorophyll absorbs specific wavelengths more than
others (the basis of its absorption spectrum). It
absorbs most visible wavelengths except in the green
part of the spectrum which, simplistically, is why
leaves appear green; it does not absorb greatly in the
far-red end of the spectrum (invisible to Man). As a
result, the light becomes enriched in far-red wave­
lengths and phytochrome is part of the response sys­
tem that responds to the red:far-red ratio. Thus, when
the seed perceives a low ratio, indicating that it lies
under another plant, it does not germinate; however,
should a gap later appear in the canopy, the red:far-red
ratio will increase, providing a signal that conditions
are appropriate for germination. A similar detection
mechanism seems to lie behind the 'acclimation'
mechanism that results in any given leaf developing
as a sun or a shade leaf.
Acknowledgements
We gratefully acknowledge the cheerful co-operation and enthu­
siastic involvement of the 1996 C class in Plant Ecophysiology
at the University of Karlstad. JDBW and HOH were supported
by the Swedish National Agency for Higher Education under
their 1996 Teacher Exchange Scheme. We thank Dr Anders
Claessen and Professor Rod Herbert for supporting the
exchange. We value Mikael Karlsson's assistance in supplying
the radish seedlings. Tracy Lawson, Richard Parsons, and John
Raven are thanked for their comments on the MS.
Appendix:
Possibilities for introducing the sun/shade leaf
response in schools
Younger secondary school students should be able to
obtain basic measurements of leaf anatomy and
growth, including most of the data collected in part 1
of table 1, though in determining the meaning of the
information collected, the group will probably need to
be led by the teacher. They should be able to feel and
describe the difference in leaf thickness in freshly
picked leaves and may be able to see colour differ­
ences when they are held up to the light (but should be
told to avoid looking at the sun). The exercise of
determining leaf area by cut-out (tracing round the
leaf outline may be appropriate if a photocopier is not
available) provides manipulative work and involves
finding an unknown from ratio analysis. The concepts
of fresh mass, dry mass, infiltrated mass and density
can be introduced or reinforced. Leaf infiltration is
possible — relatively cheap vacuum pumps operating
Journal of Biological Education (19981 32 (3) 189
 Teaching botany on the sunny side of the tree Weyers, Hoglund, and McEwen

from water taps can be used if a vacuum line is
unavailable. Drawing transverse sections of sun and
shade leaves reinforces valuable skills, though it is
recommended that sectioning by hand is carried out
by an adult or prepared slides are used (see figure 1
legend for source).
For older students, the material described here could
be used as a basis for project work in curricula which
provide an opportunity for this. Good students should
be capable of appreciating some of the finer points
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Scientific and Technical.
Larcher, W. (1995) Physiological plant ecology. 3rd edn. Berlin,
Germany: Springer.
Lawlor, D. W. (1993) Photosynthesis: molecular, physiological
and environmental processes. 2nd edn. Harlow, Essex.
Longman Scientific and Technical.
Parsons, R., Weyers, J.D.B., Lawson, T, and Godber, I. (1997)
Rapid and straightforward estimates of photosynthetic char­
acteristics using a portable IRGA system. Photosynthetica,
34,265-279.
Wintermans, J. F. G. M. and de Mots, A. (1965) Spectrophoto-

concerning evolution of the syndrome of sun and metric characteristics of chlorophyll a and b and their pheo-
shade leaves, and classroom discussion along these phytins in ethanol. Biochimica et Biophysica Acta, 109,448-
lines might provide a vehicle for deeper learning. 453.

References
Hunt, R. (1990) Basic growth indices. Cambridge: Cambridge
University Press.
Jones, A. M., Reed, R. H., and Weyers, J. D. B. (1998) Practical
skills in biology. 2nd edn. Harlow, Essex: Longman
The authors
Jonathan D. B. Weyers is a Senior Lecturer in the Department
of Biological Sciences, University of Dundee, Dundee,
Scotland DD1 4HN. Hans-Olof Hoglund is a Senior Lecturer
and Birgitta McEwen is a Lecturer in the Department of
Natural Sciences, University of Karlstad, S-651 88 Karlstad,
Sweden.

A Science Teaching Scholarship

The Hertfordshire science teaching scholarship is sponsored jointly by John Murray
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The project must be one which is intended to make the teaching of science more effective by
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190 Journal of Biological Education (1998) 32 (3)