Oecologia (Berl.

) 38, 45-50 (1979) Oecologia

9 by Springer-Verlag 1979

Carbon Isotope Ratios of Central Mexican Crassulaceae

in Natural and Greenhouse Environments

Philip W. Rundel 1,, James A. Rundel 2, H. Ziegler 3, and W. Stichler 4

Department of Ecology and Evolutionary Biology, University of California,
Irvine, CA 92717, USA
2 Sonoma World of Birds, 23570 Arnold Drive, Sonoma, CA 95476, USA
3 Institut ffir Botanik und Mikrobiologie der Technischen Universitfit,
Arcisstr. 21, D-8000 Mfinchen 2, Federai Republic of Germany
4 Institut ffir Radiohydrometrie der Gesellschaft f/Jr Strahlen- und Umweltforschung mbH,
Ingolst/idter Landstr. 1, D-8042 Neuherberg, Federal Republic of Germany

Summary. Measurements of carbon isotope ratios of central Mexican Crassu-

laceae collected over a broad habitat range show consistent patterns of
C A M activity with no indications of substantial flexible photosynthetic path-
ways between C3 and CAM. The three genera studied - Echeveria, Pachyphy-
turn, and Graptopetalum - are all closely related to Dudleya in which consider-
able flexible metabolic response has been demonstrated. Comparative mea-
surements of carbon isotope ratios for field collected and greenhouse reared
samples of the same taxa showed a uniform occurrence of slightly more
negative 6 t3C values, but no indication of substantial flexible metabolic
response.

Introduction

Considerable attention has been given in recent years to the potential ecological
advantages of flexible photosynthetic systems in succulent plants. Hypothetically,
succulent plants capable of utilizing crassulacean acid metabolism (CAM) take
advantage of the relative high water use efficiency of this system under conditions
of environmental drought. When water is not limiting, however, the higher
productivity associated with CO2 fixation via C3 metabolism should be advanta-
geous. L a b o r a t o r y studies with leaf succulents have established that a variety
of environmental conditions and genetic factors affect the ratio of COz fixed
during the day by the C3 pathway and in the dark by the C A M pathway.
Temperature (Queiroz, 1965; Neales, 1973a, b; Troughton et al., 1974), water
stress (Kluge and Fischer, 1967; Neals et al., 1968; Hartsock and Nobel, 1976),

* To whom offprint requests should be sent

0029-8549/79/0038/0045/$01.20
46 P.W. Rundelet al.

photoperiod (Lerman and Queiroz, 1974) and salinity (Winter and yon Willert,
1972) may all be important.
Field studies investigating the flexible nature of photosynthetic systems in
CAM plants have produced contrasting results. Carbon isotope ratio studies
with stem species of the Cactaceae and Agavaceae have typically showed no
consistent shift in isotope ratios with changes in aridity (Eickmeier and Bender,
1976; Szarek and Troughton, 1976). The magnitude of these values at all sites
and at each month of the year are characteristic of a strongly predominant
CAM metabolic system. Hartsock and Nobel (1976) have shown, however,
that Agave deserti can be converted to an essentially pure C3 system by artificial
watering under greenhouse conditions. Watering under field conditions produced
an intermediate condition with a mix of C3 and CAM metabolism occurring
in each diurnal cycle.
For leaf succulents, particularly those in the Crassulaceae, flexible photosyn-
thetic systems have been commonly found in field studies. This shift is particu-
larly well documented in Dudleya growing along the coast of California and
Baja California. The ratio of CO2 fixed by Ca to CAM pathway lowers over
the growing season as drought stress increases (Bartholomew, 1973). Carbon
isotope ratios of Dudleya taxa sampled along natural environmental gradients
of drought stress show gradients of change consistent with a hypothesis of
flexible photosynthetic systems (Mooney et al., 1974; Troughton et al., 1977).
Flexible photosynthetic systems have also been noted for alpine species of Sem-
pervivum from central Europe (Osmond et al., 1975).
In this paper we report measurements of carbon isotope ratios in a group
of Crassulaceae closely related to Dudleya. We have included data on 14 taxa
of Echeveria, Pachyphytum, and Graptopetalum from a wide range of thermal
and precipitation regimes in central Mexico. With a range of environmental
conditions varying from desert steppe regions of the Chihuahuan Desert through
pine-oak forests in the mountains of Hidalgo to subtropical forests in the Sierra
Mixteca of Oaxaca, evidence for possible adaptive patterns of flexible photosyn-
thetic systems should be very clear. We also report comparative carbon isotope
ratios of cultivated plants grown under non-water limited conditions.
The genus Echeveria includes approximately 150 species of succulent rossette
plants, extending geographically over a 6,000 km range from southwestern Texas
to northwestern Argentina. Within this range species occur in a wide range
of plant communities from arid Chihuahuan Desert and mesquite grassland
communities through temperate pine-oak and boreal forest associations to tropi-
cal evergreen and cloud forests. With very few exceptions. Eeheveria taxa have
highly localized distributions, characteristically inhabiting rocks, cliffs, lava
flows, or steep slopes. Root systems are usually poorly developed and subsoil
structure appears to be of minor importance. Related species occur indifferently
on basalt, rhyolite, breccias, and limestone (Walther, 1972). In these habitats
their most common associates are xerophytic ferns and species of three other
genera of Crassulaceae Sedum, Pachyphytum and Graptopetalum.
Echeveria, Pachyphytum, Graptopetalum and Dudleya are all members of
the subfamily Echeverioideae of the Crassulaceae. While most taxa of Dudleya
were once placed in Echeveria, these two genera are now considered only dis-
Carbon Isotope Ratios of Mexican Crassulaceae 47

tantly related within the subfamily (Walther, 1972) and may have had separate
origins in the Sedoideae. Pachyphytum and Graptopetalum have close affinities
with Echeveria.

Materials and Methods

Sample collections of plants for analysis were field collected in central Mexico in March 1976.
Cultivated plants of these taxa were grown a minimum of i2 months under greenhouse conditions
in moderately rich soils without fertilizer in Santa Rosa (Sonoma County) California. From mid-
March to late October they were watered one to two times per week. Extreme summer temperatures
in the greenhouse reached 40~ C. For the remaining months the pots were soaked to saturation
every 4-6 weeks but subsequently allowed to dry out completely before the next watering. This
regime provides maintenance of natural growth morphologiesof these taxa. Climatic data described
below for field sites is Viv6 and G6mez (1946). Air-dry leaf samples were shipped to Germany
by air for analysis. Analysis techniques have been previously described (Osmond et al., 1975).

Results and Discussion

Field collected taxa of Echeveria, Pachyphytum, and Graptopetalum are represen-

tative of three groups of climatic types under the K6ppen system of classification.
The first of these is the desert steppe climate type (BS). Echeveria agavoides
from near Balneario de Lourdes (San Luis Potosi) represents the most arid
extreme sampled, with a mean annual precipitation of approximately 360 mm.
Peak levels of precipitation occur in August and September. These plants grow
on dry and rocky, north-facing slopes in association with Hechtia, Bursera and
Mammillaria. Mean temperatures for San Luis Potosi, 25 km north at the same
elevation (1,900 m) are 13 ~ in January and 20 ~ in July. A similar desert steppe
environment is present at the habitats of Echeveria sanchez-mejorandae, E. bifida
and Pachyphytum bracteosum at 2,200-2,400 m in the mountains of Hidalgo
near Barranca de Mezquititlan. Thorn scrub vegetation dominates this area
with succulents present on rocky soil and on soil pockets in boulders. Precipita-
tion is slightly in excess of 400 mm y r - 1 but mean July temperatures are 4 ~ C
cooler than at San Luis P6tosi. A third desert steppe habitat near Monterey,
Nuevo Le6n, represents hotter but less arid conditions within the Chihuahuan
Desert. January and July mean temperatures are 14~ and 27 ~ C, respectively,
while precipitation is slightly over 700 m m y r - 1.
Humid temperate climates with year-round moisture availability (Type Cf)
are represented by a group of taxa collected on the Mazatlan-Durango highway
near the Sinaloa-Durango border at 2,500 m elevation. Echeveria dactylifera,
E. affinis, and Graptopetalum amethystinum grow here in pockets of leaf mold
on steep rocky slopes in a pine-oak forest. Precipitation is approximately 925 mm
yr -1. July mean temperatures are 15~ but January means drop to a cool
6~
The third K6ppen climatic regime represented in our samples is humid
temperate climate with summer precipitation (Type CW). Samples of Pachyphy-
turn glutinicaule came from the type locality along the Rio Tula in Hidalgo
48 P.W. R u n d e l et al.

Table 1. C o m p a r a t i v e 613C values of field collected a n d c u l t i v a t e d t a x a of Echeveria, Pachyphytum

a n d Graptopetalum in r e l a t i o n to K 6 p p e n climatic r e g i m e of their n a t u r a l h a b i t a t s in Mexico.
K 6 p p e n a b b r e v i a t i o n s are as folldws: BS. arid steppe; C f - h u m i d t e m p e r a t e with y e a r - r o u n d precipi-
tation; CW. humid temperate with summer precipitation

Species Location K6ppen Field Cultivated

Climatic ~13C [~ ~513C [~
Regime

Echeveria agavoides Lem. San Luis Pdtosi BS -12.88 - 14.38

Eeheveria sanchez-mejorandae W a l t h e r Hidalgo BS - 14.77 - 15.28
Echeveria bifida (Hemsl.) Schlecht. Hidalgo BS - 12.29
Pachyphytum bracteosum Link, Hidalgo BS - 14.20
K l o t s c h and O t t o
Echeveria sp. nov. (Series Urceolate) Nuevo Ledn BS - 13.66
Echereria simulans Rose Nuevo Le6n BS - 14.67
Graptopetalum amethystinum Durango Cf - 10.82 - 15.39
(Rose) W a l t h e r
Echeveria affinis W a l t h e r
- brown form Durango Cf - 13.32 - 13.88
- green f o r m Durango Cf - 12.80
Eeheveria daetylifera W a l t h e r Durango Cf - 11.51
Paehyphytum glutinieaule M o r a n Hidalgo CW - 13.57
Echeveria secunda Booth. v a t eornuta Hidalgo CW - 15.54
Eeheveria pulidonis W a l t h e r Hidalgo CW - 13.76 - 13.53
Eeheveria longissima W a l t h e r Puebla CW - 13.88
Eeheveria leucotrieha J.A. P u r p u s Puebla CW - 12.70

where the species grows on steep rock outcrops in a Lemaireocereus scrub

community. Echeveria secunda vat. cornuta comes from Barranca de Marmoles
in Hidalgo where it grows on eroded rocky slopes in a pine-oak forest. Finally,
E. pulidonis was also collected in Hidalgo in Barranca de Alamo near Carpinteros
where it grew at the bottom of the barranca on vertical granite faces. While
climatic data for the mountains of Hidalgo are poor, precipitation is evidently
in excess of 1,300 mm at the 2,200-2,500 m elevational range where these species
grow. Seasonal temperature means are comparable to values described above
for desert steppe portions of the mountains of northern Hidalgo at similar
elevations. The last two species considered, Echeveria longissima and E. leucotri-
cha come from 2,000-2,500 m in the Sierra Mixteca in southern Pueblo where
the mean annual precipitation is probably in the range of 800-1,000 mm y r - 1.
Comparative 613C values of these field collected plants, shown in Table 1,
demonstrate no correlation with climatic regime. These values ranged from
a high of-10.82~ in Graptopetalum amethystinum to a low of -15.54~
for Echeveria secunda var. cornuta. This narrow range of values all correspond
to metabolic systems which rely heavily or entirely on CAM for CO2 fixation.
No intermediate values suggestive of facultative use of both C3 and CAM
pathways of fixation (ca. - 1 6 to - 2 0 % o ) are present.
In order to investigate the potential flexibility of metabolic systems under
greenhouse conditions, ~1~C values were determined for seven cultivated plants.
While growth regimes in cultivation provided a relatively dry winter regime,
Carbon Isotope Ratios of Mexican Crassulaceae 49

these conditions were determined by experience to produce optimal growth

activity. Plants o f Pachyphytum glutinicaule and Echeveria secunda var. cornuta
were grown for one year in cultivation following field collection. Since outer
leaves were used in analyses, their 613C values p r o b a b l y closely approximate
natural field values. Cultivated plants o f Echeveria agavoides were grown for
two years following collection f r o m the same population used for the field
sample. Cultivated plants o f the other four taxa are of uncertain origin and
have spent their entire plant careers in greenhouses.
In each case where comparative data are available, 313C values are m o r e
negative in cultivated plants. While this decrease is significant no intermediate
isotope values indicative o f flexible metabolic systems were found. Unlike Dud-
leya their close relative Echeveria, Pachyphytum and Graptopetalum remained
obligate C A M plants u n d e r the non-water-limited greenhouse treatements used
in our study.
Ecological information on Echeveria are limited, but available data support
our findings. Meinzer and Rundel (1973) were able to find only C A M fixation
o f CO2 in cultivated plants o f Echeveria pumila. Echeveria columbiana growing
in subalpine and alpine p a r a m o communities up to 4,000 m elevation near
Merida, Venezuela, has a b13C value o f - 1 2 % 0 (Medina, 1976, personal c o m m u -
nication).
O n the basis o f field data alone, it might be suggested that the lack of
sensitivity of 613C values to climatic regime might relate to the xeric microhabi-
tats favored by most Crassulaceae in central Mexico. U n d o u b t e d l y m a n y steep
rock faces provide microhabitats with moisture stress conditions for plant growth
regardless o f macroclimate. The lack o f intermediate c a r b o n isotope ratios in
cultivated plants m a y indicate that these taxa are m u c h less flexible in their
metabolic p a t h w a y s than the related genus Dudleya.

Acknowledgements. The authors thank Sr. Felipe Otero for his help in carrying out our field collec-
tions. This work was completed while the senior author was an Alexander Von Humboldt Stiftung
fellow at Lehrstuhl fiJr Botanik II der Universit~it Wfirzburg, Federal Republic of Germany.

References

Bartholomew, B. : Drought response in the gas exchange of Dudleyafarinosa (Crassulaceae) grown

under natural conditions. Photosynthetica 7, 114-120 (1973)
Eiekmeier, W.G., Bender, M.M.: Carbon isotope ratios of crassulacean acid metabolism species
in relation to climate and phytosociology. Oecologia (Berl.) 25, 341-347 (1976)
Hartsock, T.L., Nobel, P.S.: Watering converts a CAM plant to daytime CO2 uptake. Nature
262, 574-576 (1976)
Kluge, M., Fischer, K.: Uber Zusammenhfinge zwischen dem CO2-Austausch und der Abgabe
von Wasserdampf durch Bryophyllum daigremontanum Berg. Planta (Berl.) 77, 212-223 (1967)
Lerman, J.C., Queiroz, O.: Carbon fixation and isotpe discrimination by a crassulacean plant:
dependence on photoperiod. Science 183, 1207-1209 (1974).
Medina, E., Delgado, M. : Photosynthesis and night CO2 fixation in Echeveriacolumbianav. Bellnitz.
Photosynthetica 10, 155-163 (I976)
Meinzer, F.C., Rundel, P.W. : Crassulacean acid metabolism and water use efficiency in Echeveria
pumila. Photosynthetica 7, 358-364 (1973)
50 P.W. Rundel et al.

Mooney, H.A., Troughton, J.H., Berry, J.A.: Arid climates and photosynthetic systems. Carnegie
Inst. Wash. Yearbook 73, 793 805 (1974)
Neales, T.F. : Effect of night temperature on the assimilation of carbon dioxide by mature pineapple
plants, Ananas comosus (L.) Merr. Aust. J. Bot. 26, 539-546 (1973 a)
Neales, T.F.: The effect of night temperature on CO2 assimilation, transpiration, and water use
efficiency in Agave americana L. Aust. J. Biol. Sci. 26, 705-714 (1973b)
Neales, T.F., Patterson, A.A., Hartney, V.J.: Physiological adaptation to drought in the carbon
assimilation and water loss of xerophytes. Nature 219, 469472 (1968)
Osmond, C.B., Ziegler, H., Stichler, W., Trimborn, P.: Carbon isotope discrimination in alpine
succulent plants supposed to be capable of crassulacean acid metabolism (CAM). Oecologia
(Bed.) 18, 209-217 (1975)
Queiroz, O. : Sur le metabolisme acide des Crassulaees: V-2. Action a long terme de la temperature
de jour sur les variations de la teneur en acide malique en jours courts. Physiol. Veg. 4,
323 339 (1965)
Szarek, S.R., Troughton, J.H.: Carbon isotope ratios in crassulacean acid metabolism plants.
Seasonal patterns in plants from natural stands. Plant Physiol. 58, 367-370 (1976)
Troughton, J.H., Mooney, H.A., Berry, J.A., Verity, D. : Variable carbon isotope ratios of Dudleya
species growing in natural environments. Oecologia (Bed.) 30, 307-311 (1977)
Viv6, J.A., G6mez, J.C. : Climatologia de Mexico. Instituto Pauamericano de Geografia e Historia,
Publ. No. 19 (1946)
Walther, E.: Echeveria. San Francisco: Calif. Acad. Sci. 1972
Winter, K., von WilIert, B.J. : NaCl-indazierter Crassulaceen-S~iurestoffwechsel bei Mesembryamh&
mum erystallinum. Z. Pflanzenphysiol. 67, 166 170 (1972)

Received September 25, 1978