Review

Tansley review
Blackwell Publishing Ltd

Photosynthetic flexibility and

ecophysiological plasticity: questions
and lessons from Clusia, the only CAM
tree, in the neotropics

Author for correspondence: Ulrich Lüttge

Ulrich Lüttge
Institute of Botany, Technical University of Darmstadt, Schnittspahnstrasse 3–5, D-64 287 Darmstadt,
Tel: +49 6151163700
Fax: +49 6151164630 Germany
Email: luettge@bio.tu-darmstadt.de
Received: 12 January 2006
Accepted: 14 March 2006

Contents

Summary 7 V. The environmental control of photosynthetic flexibility 13

I. The discovery of crassulacean acid metabolism 8 VI. Phenotypic plasticity: physiotypes and morphotypes 16
(CAM) in the trees of Clusia: arrival in the
limelight of international research VII. Ecological amplitude and habitat impact 16

II. Phylogeny 8 VIII. Conclusions and outlook 21

III. Photosynthetic physiotypes 10 Acknowledgements 22

IV. Metabolic flexibility: organic acid variations 12 References 22

Summary

Key words: Clusia, crassulacean acid
metabolism (CAM), ecophysiology,
neotropics, plasticity, photosynthesis,
phylogeny.
It is the aim of this review to present a monographic survey of the neotropical genus
Clusia on scaling levels from molecular phylogeny, metabolism, photosynthesis and
autecological environmental responses to ecological amplitude and synecological
habitat impact. Clusia is the only dicotyledonous genus with real trees performing
crassulacean acid metabolism (CAM). By way of introduction, a brief historical reminis-
cence describes the discovery of CAM in Clusia and the consequent increase in interest
in studying this particular genus of tropical shrubs and trees. The molecular phylogeny
of CAM in the genus is compared with that in Kalanchoë and the Bromeliaceae. At
the level of metabolism and photosynthesis, the great plasticity of expression of
photosynthetic physiotypes, i.e. (i) C3 photosynthesis, (ii) CAM including CAM idling,
(iii) CAM cycling and (iv) C3/CAM-intermediate behaviour, as well as metabolic flexibil-
ity in Clusia is illustrated. At the level of autecology, the factors water, irradiance and
temperature, which control photosynthetic flexibility, are assessed. The phenotypic plas-
ticity of physiotypes and morphotypes is described. At the level of synecology, the
ecological amplitude of Clusia in the tropics and the relations to habitat are surveyed.

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8 Review Tansley review
I. The discovery of crassulacean acid metabolism
(CAM) in the trees of Clusia: arrival in the
limelight of international research
As we shall see, species of Clusia are handsome woody plants.
Some have small flowers only, but others develop rather
large beautiful flowers; for example, the flowers of Clusia
grandiflora Engl. have a diameter of 150 mm. Nevertheless,
Clusia most likely would have remained just one genus among
a vast number of others in the large biodiversity of trees and
shrubs in the tropics if it had not turned out that species of
Clusia are performing crassulacean acid metabolism (CAM).
This mode of photosynthesis is basically characterized by
nocturnal uptake of CO2 and dark fixation via phosphoenol-
pyruvate carboxylase (PEPC), where the resulting fixation
product, mainly malic acid, is stored in the central cell sap
vacuole. Behind closed stomata and in the absence of any
overt gas exchange, the organic acid is remobilized during
the light period and decarboxylated, and the CO2 regained
is refixed via ribulose-bis-phosphate carboxylase/oxygenase
(Rubisco) and assimilated in the Calvin cycle.
Clusia is the only genus of trees with this mode of photosyn-
thesis. True, there are a number of sizeable plants with CAM
in the families of Cactaceae, Euphorbiaceae and Didieraceae
as well as the monocotyledonous Yucca, all of which have
been considered ‘fantastic trees’ (Menninger, 1967; Ellenberg,
1981). Moreover, giant cactus communities in Venezuela have
been called ‘cactus forests’ (Vareschi, 1980). However, Clusia
is the only genus of trees sensu stricto with typical dicotyledo-
nous secondary growth that performs CAM.
Clusia early raised the curiosity of researchers because of the
peculiarities of its photosynthetic physiology. During field
work in February 1800 in Venezuela, Alexander von Hum-
boldt discovered that Clusia rosea Jacq. had no obvious overt
gas exchange in the light period but built up a high internal
gas pressure with an oxygen concentration near 40% (Krätz,
2001; Lüttge, 2002). In 1937, Willy Hartenburg reported
observations from glasshouse experiments showing that on
bright days Clusia mexicana Vesque did not take up CO2 in
the light period but even released some CO2, which was not
explicable by respiration. However, on overcast days the CO2
exchange pattern was as expected of photosynthesizing plants
(Hartenburg, 1937; Lüttge, 1995b). Neither Alexander von
Humboldt nor Willy Hartenburg was able to really explain
their observations. Now, as we know that Clusia species are
performing CAM, we realize that they saw particular features
of CAM, where stomata are closed in the light period when
CO2 remobilized from nocturnally stored organic acids is
photosynthetically assimilated, which is associated with the
build up of high internal partial pressures of oxygen (Spalding
et al., 1979). Decarboxylation of nocturnally stored organic
acids behind closed stomata also leads to high internal CO2
concentrations, i.e. 2–60 times atmospheric (Lüttge, 2002),
which may even cause some diffusive loss of CO2 to the
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atmosphere in spite of stomatal closure. It remained for the
Mexicans Tinoco Ojanguren & Vazquez-Yanez (1983) to
demonstrate and clearly explain the performance of CAM in
Clusia for the first time, and with an additional seminal study
by Ting et al. (1985) this became widely known and created
wide interest, moving Clusia into the limelight of interna-
tional research activities. This now allows me to present Clusia
in this review, and we are also preparing a monographic book
on Clusia covering all aspects of its biology, i.e. its taxonomy
and phylogeny, diversity and phytogeography, ecology and
physiology, and circadian clock (Lüttge, 2006c).
II. Phylogeny
The question is why there are not more trees with CAM.
Similarly, although there are woody plants, shrubs and small
trees with C4 photosynthesis, there are no real large trees with
this mode of photosynthesis. The two modes, C4 and CAM,
share many biochemical features, particularly the primary
fixation of CO2 by PEPC and synthesis of organic acids
(mainly malate and citrate in CAM and malate and aspartate
in C4 photosynthesis) and the subsequent decarboxylation
of the organic acids and refixation and assimilation of the
CO2 by Rubisco in the Calvin cycle. There are, of course,
important differences between C4 photosynthesis and
CAM which are well known and need not be reiterated here.
However, the observation that trees do not use phosphoenol-
pyruvate (PEP) carboxylation as widely as CAM and C4
photosynthesis are otherwise distributed among angiosperms
shows an intriguing parallel between both modes of photo-
synthesis. Phylogenetic studies address the question of why
this is so, but we must note here, at the outset, that in both
cases the question has not been answered to date.
CAM is widespread among the taxa of vascular plants
including ferns and fern allies (Isoëtes). There is no doubt that
CAM arose polyphyletically many times during the evolution
of vascular plants (Lüttge, 1987). This must have been facilitated
by the fact that CAM does not require any particular metabolic
trait as compared with non-CAM plants. A reorganization
and specific management of metabolic modules common in
the housekeeping functions of all photosynthesizing plants
are sufficient for CAM performance (Lüttge, 2005). The
evolution of CAM has been specifically studied in the genus
Kalanchoë and the family Bromeliaceae and more recently also
in the genus Clusia. There are conspicuous differences among
the three taxa. A comparison (Lüttge, 2004, 2005) is useful as
it reveals interesting peculiarities of Clusia.
The radiation centre of Kalanchoë is in the moist forests of
eastern Madagascar (Kluge et al., 1991; Gehrig et al., 2001;
Kluge, 2005). The genus comprises species with obligate C3
photosynthesis and obligate CAM and there are a small
number of species that are C3/CAM intermediate and can
switch between the two modes of photosynthesis (Brulfert
et al., 1973, 1975). The majority of the species in the genus

are CAM species. Morphological (Boiteau & Allorge-Boiteau,
1995) as well as molecular (Gehrig et al., 1997, 2000, 2001)
evidence suggests that CAM must have evolved monophylet-
ically in the genus from a C3-photosynthesis ancestor during
radiation from the moist east to the drier areas up to the arid
areas in the west and south-west of Madagascar, where CAM
functions as an important adaptation to the limited availability
of water.
Conversely, in the neotropical family of the Bromeliaceae
with its three subfamilies Pitcairnioideae, Bromelioideae and
Tillandsioideae and very many epiphytic species, again as
supported by morphological/anatomical, biochemical and
molecular genetic evidence (Smith, 1989; Crayn et al., 2004),
CAM and the epiphytic habit evolved a minimum of three
times. Epiphytism and CAM have evolved independent of
each other, although evidently CAM is an adaptation to the
water shortage stress of the epiphytic habitat (Zotz & Hietz,
2001). The last common ancestor of the Bromeliaceae was a
terrestrial C3 mesophyte. It is important to note that there has
also been a reversion from CAM back to C3 photosynthesis
in some taxa of the bromelioid line following subsequent
radiation into less xeric habitats (Crayn et al., 2004). Diversity
is very large in the family in many respects but photosynthetic
plasticity is low. There are obligate C3 and CAM species but
in the entire large family there appears to be only one clearly
C3/CAM-intermediate species, namely Guzmania monos-
tachia (L.) Rusby ex Mez (Maxwell et al., 1994, 1995, 1999;
Maxwell, 2002).
The subfamily Clusioideae with the genus Clusia in the
family Clusiaceae has a minimum phylogenetic age of
90 × 106 years (Gustafsson et al., 2002). Clusia is a large
genus of shrubs and trees with about 300–400 species (Pipoly
et al., 1998). The genus is considered monophyletic. However,
internal transcribed spacer (ITS) sequencing of nuclear
ribosomal DNA has shown that the evolution of CAM in the
genus was polyphyletic (Vaasen et al., 2002; Gehrig et al.,
2003; Holtum et al., 2004). ITS analyses of some 80 species
of Clusia and a derived phylogenetic tree suggest that CAM
evolved at least twice in the genus. A reversion from CAM to
C3 was noted in eight individual species and in one group of
three species (Gustafsson et al., 2006). Thus the situation in
Clusia resembles that in the Bromeliaceae and not that in
Kalanchoë. However, in contrast to the Bromeliaceae, Clusia
is characterized by an enormous plasticity. There are very
many C3/CAM-intermediate species. It appears that this
even may be the rule in the genus and obligate C3 and especi-
ally obligate CAM species are much fewer (Holtum et al.,
2004).
This expectation is, however, based on a much smaller
sample size. To some extent, the occurrence of CAM can be
screened by analysing carbon isotope ratios (δ13C) because, as
a result of the much lower 13C discrimination of PEPC (−7‰
relative to CO2) than of Rubisco (+27‰ relative to CO2)
(Ziegler, 1994), in CAM-performing plants δ13C values are
© The Authors (2006). Journal compilation © New Phytologist (2006) www.newphytologist.org
Tansley review Review 9
significantly less negative than in C3 plants. However, it has
been noted that C3/CAM-intermediate species often make
little use of their CAM option in the field, so that δ13C values
are C3-like. Although Winter & Holtum (2002) and Winter
et al. (2005) have presented a calibration for deducing from
δ13C values the extent of primary dark fixation (PEPC) and
light fixation (Rubisco) of CO2, respectively, for a definitive
identification of a CAM potential in a species additional
physiological evidence is required, for example from gas
exchange measurements and analyses of diurnal organic acid
fluctuations. It is much more tedious to obtain such informa-
tion for many species than to just analyse δ13C values, but it
appears to be very important to do this for as many species of
Clusia as possible. This would not only provide insights into
the evolution of CAM within the genus. In addition, because
of the extraordinary inherent evolutionary plasticity in the genus
and its high speciation rate (Gustafsson et al., 2006), the
investigation of Clusia may also advance understanding of
CAM evolution in general.
In summary, the comparison of the three taxa Kalanchoë,
Bromeliaceae and Clusia, in which CAM phylogeny has been
studied in the greatest depth, shows that the evolution of
CAM not only occurred independently in the three taxa but
also went rather different ways, i.e. monophyletically within
the genus Kalanchoë, polyphyletically but without genera-
tion of much plasticity in the Bromeliaceae and polyphylet-
ically with production of enormous plasticity in the genus
Clusia.
The key enzyme of CAM as well as C4 photosynthesis is
PEPC. Recently, the molecular characterization of PEPC
isoforms has been investigated in detail in C4 photosynthesis
(Engelmann et al., 2003; Svensson et al., 2003; Westhoff &
Gowik, 2004) and also in CAM plants, with a strong phylo-
genetic bias. It has been found that Kalanchoë pinnata (Lam.)
Pers. has seven PEPC isoforms, only one of which is CAM
relevant (Gehrig et al., 2005). Taybi et al. (2004) studied the
expression of PEPC and PEPC-kinase (PEPCK ) genes in four
different species of Clusia. PEPCKs regulate the activity of
PEPC which is more active and less sensitive to feedback
inhibition by malate in the phosphorylated state. The species
compared were C. rosea, an obligate CAM species, Cluisa
minor L., a C3/CAM-intermediate species, Clusia aripoensis
Britt., a weakly CAM-inducible species, and Clusia multiflora
H.B.K., an obligate C3 species. They found that transcriptional
control of PEPC abundance is a key factor in determining the
genotypic capacity of CAM, while the PEPCK gene is under
the control of metabolic feedback, and there are no C3- or
CAM-specific isoforms of the PEPCK genes. The obligate
CAM species Clusia hilariana Schlecht. produces three
isoforms of PEPC, one of which is root-specific and is
probably a housekeeping isoform in this organ, while the C3
species C. multiflora and the C3/CAM-intermediate species
C. minor only have one isoform (Vaasen, 2005; Vaasen et al.,
2006). For C. minor the observation that there is only one
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10 Review Tansley review
isoform each of PEPC and PEPCK is most noteworthy
because this implies that just one isoform can support both
the housekeeping functions of the C3 state and the particular
requirements of the CAM state. This may facilitate C3–CAM
switches. PEPC transcript abundance is increased in the
CAM state (Taybi et al., 2004; Vaasen, 2005; A. Vaasen, D.
Begerov, R. Hampp, Botanisches Institut, Universität Tübingen,
Tübingen, Baden-Württemberg, Germany). The observation
that a single isoform each of PEPC and PEPCK can support
performance of both C3 photosynthesis and CAM also
strengthens the speculation that this has facilitated CAM
evolution and that C3/CAM flexibility, which is so wide-
spread in the genus Clusia, has played a role in this. However,
whether the absence of more isoforms in C. minor is a result
of a lack of evolution of more isoforms or the loss of redun-
dant isoforms during evolution remains unresolved at
present. More comparative work clearly is highly desirable.
However, in the earlier literature it was not C3/CAM-
intermediate behaviour but ‘CAM cycling’ that was suggested
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to have been a starting point for CAM evolution (Guralnick
et al., 1986; Guralnick & Jackson, 2001), but this now takes
us to the next section discussing photosynthetic plasticity in
Clusia.
III. Photosynthetic physiotypes
In Fig. 1, I have reduced metabolic complexity to the extent
of only using two modules, namely PEPC and organic acids
and Rubisco and carbohydrate ([CH2O]n), respectively, and
external and internal CO2 for a schematic illustration of four
different photosynthetic physiotypes, all of which are found
among Clusia species.
1 C3 photosynthesis, where stomata are open during the light
period and CO2 taken up is fixed directly via Rubisco in the light.
2 CAM, where stomata are open in the dark period and CO2
taken up is fixed via PEPC leading to the production of
organic acid (both malic and citric in Clusia species), organic
acid is decarboxylated during the light period and the CO2
Fig. 1 Minimal scheme of metabolism
using two metabolic modules, namely
phosphoenolpyruvate carboxylase (PEPC)
and nocturnal organic acid accumulation
(org.ac.) and ribulosebisphosphate
carboxylase/oxygenase (Rubisco) and
daytime carbohydrate synthesis ([CH2O]n)
and two pools of inorganic carbon, namely
external and internal CO2, to illustrate the
basic features of four photosynthetic
physiotypes and their connections: from left
to right, (i) crassulacean acid metabolism
(CAM) cycling (CAM cycl), (ii) full CAM with
CAM idling (CAM idl), (iii) C3/CAM-
intermediate behaviour, and (iv) C3
photosynthesis. Dark arrows, carbon flow in
the dark; white arrows, carbon flow in the
light.
 Tansley review Review 11

regained is refixed via Rubisco. Some direct C3-type fixation

is also possible, i.e. in the later light period, when ecological
conditions, particularly water relations, allow stomatal opening
after nocturnally accumulated organic acid is consumed.
CAM idling, as also shown in Fig. 1, is not considered a
separate physiotype but a variant of normal full CAM, where
stomata remain closed night and day when water relations are
adverse and respiratory CO2 is refixed in the dark period and
recycled to carbohydrate during the light period.
3 C3/CAM-intermediate behaviour with reversible switches
between the two modes of photosynthesis in perennial Clusia,
where leaves are used for at least two growth periods (Olivares,
1997).
4 CAM cycling, where CO2 uptake and fixation occur
mainly during the light period as in C3 photosynthesis, but
respiratory CO2 is recuperated via PEPC behind closed stomata
during the dark period and organic acid is produced which is
decarboxylated during the light period, supplementing the CO2
taken up from the atmosphere via the open stomata as substrate
for Rubisco. This use of PEPC to prevent loss of respiratory
CO2 in the dark period was thought to have been an early step
towards the evolution of CAM, as alluded to in the previous
section (Guralnick et al., 1986; Guralnick & Jackson, 2001).
The scheme illustrated in Fig. 1 symbolizes the modular
nature of photosynthetic physiotypes and can be used to
elucidate the links that might be important in long-term and
medium-term evolutionary and ontogenetic development,
respectively, as well as in short-term environmental responses
in ecophysiology. For example, (i) CAM can also involve a
strong C3 element; (ii) without light-period CO2 uptake
CAM cycling would become CAM idling.
Some representative curves of day:night CO2 exchange in
Clusia species are shown in Fig. 2. A bona fide obligate C3
species is C. multiflora and obligate CAM species are Clusia
alata Pl. et Tr., Clusia major L., C. rosea and C. hilariana. As
noted in the previous section, there are very many C3/CAM-
intermediate species in the genus Clusia. The most widely Fig. 2 CO2 gas exchange curves ( JCO ) of Clusia minor L. under
2
studied species in this respect is C. minor. Under different condi- three different conditions (A–C) and Clusia venosa Jacq. (D), Clusia
alata Pl. et Tr. (E) and Clusia major L. (F), showing performance of C3
tions with respect to a combination of different environmental photosynthesis (A, D), crassulacean acid metabolism (CAM) (B, E, F),
factors, such as irradiance during growth and a variety of with the four phases of (I) nocturnal CO2 uptake, (II) daytime
day:night temperature regimes, it produced a vast number stomatal closure and organic acid remobilization, (III) a transition with
of different responses including full CAM with CAM idling CO2 uptake in the early light period, and (IV) CO2 uptake in the later
(Haag-Kerwer et al., 1992). In another experiment, irradiance light period (Osmond, 1978), and CO2 uptake almost around the
clock and strong expression of all four CAM phases (C). In (A–C) for
during growth and nitrogen nutrition were varied. Plants C. minor, the irradiance and water vapour pressure deficit (VPD) of
grown at high irradiance without supplemental nitrogen the atmosphere in µmol m−2 s−1/mbar bar−1, respectively, were as
showed no CO2 uptake and a minimal loss of CO2 but accu- follows: (A) 1700/6.6, (B) 400/13.5, (C) 400/3.5, i.e. high irradiance
mulated appreciable amounts of citrate plus a small amount of and low VPD favoured C3 photosynthesis, medium irradiance and
malate in the dark period, while light-period CO2 uptake was high VPD favoured CAM, and medium irradiance and very low VPD
favoured CO2 uptake around the clock. Dark bars, night-time; white
substantial, i.e. there were clear indications of CAM cycling bars, daytime. (After data of Lee et al., 1989, where C. minor is
(Franco et al., 1991). Most of the experiments cited here wrongly named C. rosea, and Franco et al., 1990.)
were performed with one clone of vegetatively propagated
C. minor. The enormous phenotypic plasticity with respect to
expression of photosynthetic physiotypes of the genome is
evident.

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12 Review Tansley review
IV. Metabolic flexibility: organic acid variations
Clusia also shows flexibility at the level of metabolites in the
CAM cycle. Normally the precursor for glycolytic formation
of phosphoenolpyruvate (PEP) as a CO2 acceptor for dark
fixation is glucan (starch) and the acid produced is malic acid
(malate). In Clusia, however, free sugars, i.e. glucose, fructose
and sucrose, are also used in addition to starch as PEP
precursors (Popp et al., 1987; Ball et al., 1991; Berg et al.,
2004). Although noteworthy, this situation is not unique as it
also occurs occasionally in other taxa, for example in the
bromeliad pineapple Ananas comosus (L.) merr. (Black et al.,
1996). More remarkable is the fact that Clusia, in addition or
alternatively to malate, can also synthesize citrate. There are
other CAM plants that nocturnally form citrate in their CAM
cycle but the amounts are far lower than those found in Clusia;
for example, day:night oscillations of citrate concentrations
in some species of Kalanchoë may be up to 26 mM, and this
is considered relatively high (Lüttge, 1988), but C. minor
and C. rosea can show values of up to 200 mM (Franco et al.,
1992). This really amounts to a qualitative rather than only a
quantitative difference. The relative amounts of malate and
citrate produced can vary within the same species, clone or
plant depending on environmental conditions, where
again C. minor is an excellent example (Haag-Kerwer et al.,
1992).
I can list three cases where differential responses of
nocturnal malate and citrate accumulation in the CAM of
Clusia species to environmental factors have been observed.
1 Under drought conditions in experiments by Franco et al.
(1992) and de Mattos et al. (1999) with three Clusia species
(Clusia lanceolata Camb., C. rosea and C. minor) the ratio of
malate to citrate accumulation declined by a factor of 1.8–3.4
after 10 to 16 d without watering. This was, however, not
observed in a study by Borland et al. (1998) with C. minor,
C. rosea and C. aripoensis, but in this case the ratios of malate
to citrate were already very low in the nondroughted controls.
2 In C. minor the ratio of malate to citrate accumulated also
depended on nitrogen nutrition and irradiance during growth
Table 1 Potential advantages and disadvantages, respectively, of night:day change in the concentrations of malate (∆mal ) and citrate (∆citr )
during the crassulacean acid metabolism (CAM) cycle (Franco et al., 1992; Lüttge, 1996, 2006a)
Function ∆mal
1. CO2 acquisition Yes
2. H2O saving during CO2 acquisition Yes
3. Carbon recycling in the dark period CO2
4. Carbon recycling in the light period and CO2
increase in internal CO2 concentration Yes
5. Energy budget in the dark period ATP consumption
6. Energy budget in the light period ATP consumption
7. Osmotic changes Yes
8. Buffer capacity Low
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(Franco et al., 1991). At low irradiance there was no malate
and only citrate accumulation with and without supplemental
nitrogen nutrition. At high irradiance the ratio of malate to
citrate was 0.22 without and 3.07 with nitrogen.
3 In C. minor the ratio of malate to citrate was also affected
by day:night (D:N) temperature regimes and irradiance
during growth. For example, low irradiance and D:N tem-
peratures of 30 : 15°C produced only malate accumulation;
high irradiance and D:N temperatures of 25 : 15°C or 30 : 15°C
produced high malate and low citrate accumulation; high
irradiance and D:N temperatures of 25 : 20°C produced
equal malate and citrate accumulation; low irradiance and D:N
temperatures of 30 : 30°C produced only citrate accumulation;
high irradiance and D:N temperatures of 20 : 20°C produced
high citrate and low malate accumulation (Haag-Kerwer
et al., 1992).
Naturally one would expect that a switch in metabolism as
important as a change from malate to citrate accumulation
should have adaptive benefits in response to the various
environmental cues. However, the picture created by the
above three examples of environmental responses remains
very fuzzy, and we can only try to evaluate comparatively a
number of roles of malate and citrate, respectively, which I
summarize below in eight points (Table 1; Franco et al., 1992;
Lüttge, 1996, 2006a).
1 With respect to day:night carbon balance, the CAM cycle
with citrate is futile. Nocturnal citrate accumulation does not
contribute to carbon gain because, starting with a C6-hexose
unit as precursor, the result is C6-citrate. Conversely, two
molecules of C4-malate are formed by nocturnal fixation of
2 CO2 per C6-hexose unit used (Lüttge, 1988).
2 As it does not contribute to carbon gain, citrate, of course,
also does not contribute to water saving during CO2 acquisition,
unlike malate synthesis by nocturnal CO2 uptake when
evaporative demand is much lower than during the day.
3 Like malate, citrate can contribute to carbon recycling
during the dark period, for example in CAM idling. With
malate accumulation respiratory CO2 is recycled; with citrate
accumulation entire carbon skeletons are recycled.
∆citr
No
No
Carbon skeleton
CO2
Yes, more than for malate
ATP consumption and production of redox power
Larger ATP consumption and consumption of redox power
Yes, less than for malate
High

4 Like malate, citrate can contribute to CO2 recycling in the
light period. Potentially more CO2 is generated from citrate
breakdown in the light period so that citrate can make a larger
contribution to the build-up of high internal CO2 concentrations
behind closed stomata than malate.
5 Energetically, citrate accumulation is marginally cheaper in
the dark period. ATP is consumed in similar amounts in both
malate and citrate accumulation but reduction equivalents
are generated in citrate synthesis which can be used for ATP
production in the respiration chain.
6 Energetically, the return of carbon from citrate to carbohy-
drate in the light period appears to be more costly than in the
case of malate. The decarboxylation of two molecules of
malate gives rise to two molecules of pyruvate, which can be
used to regenerate two molecules of triose and then one
molecule of hexose via gluconeogenesis. The decarboxylation
of one molecule of citrate may generate three molecules of
CO2 and only one molecule of pyruvate. The refixation of
CO2 to generate triose is much more costly energetically than
gluconeogenesis starting from pyruvate.
7 The nocturnal accumulation of malate in the vacuoles of
CAM plants has osmotic consequences and can support
nocturnal water storage (Eller & Ruess, 1986; Lüttge, 1986;
Ruess et al., 1988; Eller et al., 1992; Murphy & Smith, 1998).
The CAM cycle with citrate is less effective in this respect; for
example, for one hexose unit from glucan, two osmotically
active malate molecules and only one citrate molecule are
formed. If the precursor is free hexose, citrate formation is
osmotically neutral.
8 Citrate is a more effective pH-buffer compound than
malate. The buffer capacity of the vacuole is important for
nocturnal organic acid accumulation because it determines
the trans-tonoplast proton gradient against which the proton
pumps energizing acid accumulation must work (Lüttge
et al., 1981). A higher buffer capacity may therefore allow an
increased capacity for organic acid accumulation (Franco
et al., 1992). Thus, citrate accumulation in Clusia species may
be an explanation of the fact that in Clusia species we observe
the highest nocturnal accumulation of acids ever found
among CAM plants, namely 1410 mM titratable protons in
one example of C. minor in the field in Trindiad (Borland
et al., 1992).
Hence, there is no benefit from citrate accumulation for
carbon gain and improvement of water relations (points 1, 2
and 7 above). Conversely, citrate is useful as it could facilitate
a larger total accumulation of acids in the dark period (point
8) which contributes to the build-up of high internal CO2
pressures in the light period, in which citrate is more effective
than malate (point 4). This is useful because high internal
CO2 concentrations may bring Rubisco closer to substrate
saturation, or actually achieve substrate saturation, minimizing
photorespiration, photoinhibition and photodestruction at
high irradiance during the light period (Lüttge, 2002). The
higher energy demand of citrate cycling in the light period
© The Authors (2006). Journal compilation © New Phytologist (2006) www.newphytologist.org
Tansley review Review 13
(point 6) in this respect may also be considered beneficial as
it contributes to energy dissipation, reducing the danger of
photoinhibition and photodestruction at high irradiance. In
this respect, observations in the field in Trinidad are interesting,
where citrate breakdown in C. minor during the day showed
a much closer correlation with irradiance as compared with
malate (Borland et al., 1996). Thus, one might expect that
citrate accumulation might be more favourable than malate
accumulation under conditions of drought, when stomata
partially or fully close during the dark period and internal
carbon recycling and CAM idling become increasingly
important, and also because drought is often associated with
high-irradiance stress. However, apart from the observation
that malate:citrate ratios during nocturnal acid accumulation
under drought conditions may decrease, the other results on
differential effects of environmental factors on malate and
citrate accumulation, respectively, presented above do not show
any obvious relations to the theoretically expected benefits
from citrate accumulation.
It is astonishing that such a fascinating problem as the
putative functional advantage of the metabolic plasticity of
performing CAM with malate or citrate is not understood.
Even information about the metabolic pathway involved in
nocturnal citrate synthesis is scarce (Olivares et al., 1993), and
we have no information on light-period citrate metabolism in
Clusia, so that we are left with known general biochemistry of
intermediate metabolism (see also Holtum et al., 2005), for
example in the assessment of energy budgets, as mentioned
above. Some classical biochemical studies would be very
interesting, and rigorously systematic comparative studies
varying environmental parameters may provide important
new insights. However, current fashions in plant biology have
a very different focus from motivating funding of the study of
such an exciting problem in biochemical ecology.
V. The environmental control of photosynthetic
flexibility
Environmental input is always received by the phenotypes,
and in the case of the different modes of photosynthesis these
are the photosynthetic physiotypes distinguished above
(Fig. 1). There is then a possible feedback to the genotype
which may respond with changes of the phenotype expressed
(Lüttge, 2005). More straightforwardly, we can ask the
questions of which environmental factors determine the
reversible switches between C3 photosynthesis and CAM in
Clusia species and how this works. As an example, we have
already seen above that quite specifically the expression of
PEPC may change during a C3 to CAM transition. The major
external factors are water, irradiance and temperature (Lüttge,
2004). In the field they never act individually but always
interactively. Even in experiments they are often hard to
separate. However, individual factors may become limiting
and we may try to consider them separately.
New Phytologist (2006) 171: 7–25
14 Review Tansley review
1. Water
Although the earliest environmental pressure for the
evolution of CAM was probably the problem of CO2
acquisition with the advantage of the 60 times higher CO2
affinity of PEPC than Rubisco and the related function of
CAM as a CO2-concentrating mechanism (Lüttge, 2002), it
is evident that the driving force for the polyphyletic evolution
of CAM in so many taxa of the vascular plants must have been
limited availability of water.
In the C3/CAM-intermediate C. minor, CAM can be
induced by drought. In an experiment where this was done it
was shown subsequently that even the two opposite leaves on
one node simultaneously could perform C3 photosynthesis
and CAM, respectively. The two leaves were kept in gas
exchange cuvettes at a low (6.2 mbar bar−1) and high
(13.1 mbar bar−1) water vapour pressure deficit (VPD) of the
air, respectively, i.e. subjecting the two leaves to different
evaporative demand. When the plant was re-watered after 4 d
of drought, the leaf in the dry air continued to perform CAM
with dominating CO2 uptake and fixation in the dark, while
the leaf in the moist air within a few hours switched to CO2
uptake in the light (Schmitt et al., 1988, where C. minor is
wrongly called C. rosea).
While this illustrates the plasticity of a single plant, K.
Winter and colleagues have tried to carry out a more general
survey of Clusia species, assessing water use efficiency (WUE)
by carbon isotope analysis (δ13C). WUE is defined as the ratio
of moles of CO2 fixed and assimilated to moles of water lost
by transpiration. WUE is high in nocturnal CO2 fixation during
CAM because CO2 acquisition in the dark occurs at low
evaporative demand. WUE is much lower in C3 photosynthesis
in the light and also in CAM plants when they make use of
their potential for direct CO2 fixation via Rubisco in the later
light period (Fig. 1). As the 13C discrimination of PEPC is so
much lower than that of Rubisco, analysis of δ13C values
allows screening (see section II) of the extent to which plants
use primary CO2 fixation via PEPC and Rubisco, respectively
(Borland et al., 1993), and allows this to be related to WUE.
Winter & Holtum (2002) and Winter et al. (2005) have
shown in general that δ13C values in CAM species strongly
correlate with WUE. These authors also screened 38 species
of Clusia from Panamá and found that under natural con-
ditions in the field Clusia species only sparingly make use of
their PEPC option of CO2 fixation (Holtum et al., 2004) and,
thus, the related high WUE. This underlines the importance
of other factors in addition to water in the habitats of Clusia
species.
2. Irradiance
High irradiance increases VPD and may cause over-
energization of the photosynthetic light-harvesting apparatus.
Thus, one would expect that it would support the use of the
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CAM option in plastic Clusia species. However, this is not
always so. In well-watered C. minor an increase of the
irradiance from 360 to 1200 µmol m−2 s−1 suppressed CO2
dark fixation (Schmitt et al., 1988). In studies of the return
from CAM to C3 photosynthesis in C. minor, de Mattos &
Lüttge (2001) found that, provided water is not limiting and
the plant can afford increased transpiration, daily photon use
increases when there is unrestricted C3-like CO2 uptake,
where the plant may overcome the limitations of the storage
capacity of the vacuole for nocturnal organic acid accumulation,
improving its daily carbon balance.
A comparative phytotron study of the obligate C3 species
C. multiflora and the C3/CAM-intermediate species C. minor
(Herzog et al., 1999) is interesting because it also sheds light
on the behaviour of the species and their relative use of the
CAM option in the field. Plants of the two species were grown
at a low daily dose of irradiance (4 mol m−2 d−1) and then trans-
ferred to two higher daily doses of 24.5 and 33.5 mol m−2 d−1,
respectively. Daily irradiance was applied in a bell-shaped
pattern as in nature, i.e. increasing from dawn towards noon
and decreasing again towards dusk. Figure 3 shows some
results obtained 5 d after the transfer. We can see that the C3
species which had maximum rates of CO2 uptake ( J CO2)
between 2 and 4 µmol m−2 s−1 at 4 mol m−2 d−1 could make
use of the higher irradiance of 24.5 mol m−2 d−1 for increasing
J CO up to 6 µmol m−2 s−1 but was strongly inhibited by the
2
still higher irradiance of 33.5 mol m−2 d−1 showing only a J CO2
of < 1 µmol m−2 s−1. Conversely, the C3/CAM-intermediate
species which had performed C3 photosynthesis with maximum
rates of J CO2 of c. 2–3 µmol m−2 s−1 at 4 mol m−2 d−1
had switched to CAM under the higher daily doses of irradiance
with high potential rates of J CO2 even in the transition phase
between nocturnal CO2 uptake and daytime stomatal closure
in the morning and during C3-like CO2 fixation in the later
part of the day, with no difference between the two high daily
doses. At 4 mol m−2 d−1 neither species was photoinhibited.
However, under the higher irradiances they were similarly
photoinhibited and increasingly so in the second part of the
light period after irradiance was increased from 24.5 to
33.5 mol m−2 d−1 as given by the potential quantum yield of
photosystem II (Fv/Fm, where Fv is the variable and Fm is the
maximum fluorescence of dark-adapted leaves and a ratio
below 0.83–0.80 indicates photoinhibition; Björkman &
Demmig, 1987). Nonphotochemical quenching (NPQ) of
chlorophyll (Chl) a fluorescence of photosystem II was also
similar for the two species, as were the levels of the xanthophyll
zeaxanthin, which is involved in harmless thermal energy dis-
sipation under irradiance stress (Hager 1980; Demmig-Adams,
1990; Demmig-Adams & Adams, 1992; Horton et al., 1994;
Pfündel & Bilger, 1994; Schindler & Lichtenthaler, 1996).
With the putative protective function of the high internal
CO2 concentrations built up during CAM in the light period
(see section IV), one would have expected that Fv/Fm would
have been higher and NPQ and zeaxanthin levels lower in the
 Tansley review Review 15

Fig. 3 Photosynthetic performance of Clusia

multiflora H.B.K. (solid lines) and Clusia
minor L. (broken lines) grown at a daily
irradiance of 4 mol m−2 d−1 5 d after transfer
to high irradiance of 24.5 and 33.5 mol m−2 d−1,
respectively, in a phytotron where irradiance
was increased up to midday and then
decreased again in a bell-shaped pattern
(not shown). JCO , CO2 exchange; Fv/Fm,
2
potential quantum yield of chlorophyll a of
photosystem II; NPQ, nonphotochemical
fluorescence quenching; Z, levels of
zeaxanthin, where data are only available for
24.5 mol m−2 d−1. Dark bars, night-time;
white bars, daytime. (After data of Herzog
et al., 1999.)

C3/CAM-intermediate species after its switch to CAM than
in the C3 species. That this is not so shows that CAM species,
like C3 species, are not exempt from photoinhibition and have
protective mechanisms, such as thermal energy dissipation, to
deal with irradiance and oxidative stress, as was also observed
in the CAM species C. hilariana (Franco et al., 1999). Measure-
ments by Winter et al. (1990) showed that in the early light
period in CAM-performing C. rosea the level of zeaxanthin
greatly increased from much lower levels maintained in the
dark period, decreased in the middle of the day when stomata
closed and organic acid was remobilized, and subsequently
increased again as stomata opened in the later light period.
The levels of violaxanthin, the double epoxide of zeaxanthin,
showed the opposite pattern during the day. With respect
to the pair of Clusia species, C. multiflora and C. minor, com-
pared by Herzog et al. (1999), the long-term comportment
after transfer to higher irradiance is important. In the C3 spe-
cies, under higher daily irradiance leaves became necrotic and
died. However, the plant was not principally suffering from
the high irradiance. It only suffered from the sudden change
from low irradiance during growth to higher irradiances,
probably because acclimation of the light-harvesting appara-
tus takes a long time or is totally insufficient. However,
C. multiflora then produced new leaves from dormant buds
which were adapted to the new situation. Conversely, the C3/
CAM-intermediate species C. minor was not damaged after
the transfer to higher irradiance. It was probably the much
more rapid switch of the biochemical machinery to CAM
combined with protective mechanisms which saved it from
destruction of its leaves. In fact, separate experiments showed
that the capacity to produce zeaxanthin was even higher in
C. minor (c. 130 µmol m−2 leaf area) than in C. multiflora
(c. 60 µmol m−2) but it did not make use of this higher capacity,
at least up to 5 d after transfer (Fig. 3).

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16 Review Tansley review
These phytotron experiments suggest that the C3/CAM-
intermediate species may have a larger niche width, performing
well and with high short-term flexibility both under full sun
exposure and in more shaded situations, while the C3 species
performs well under high irradiance only when it has long-term
development under full exposure, for example starting with
seedling growth. Indeed, in the field it has been observed that
C. multiflora occupies exposed sites in a secondary savanna
while C. minor prefers semi-shaded sites in a deciduous dry
forest but can also invade the exposed sites of C. multiflora
(Franco et al., 1994; Grams et al., 1997).
3. Temperature
It is widely assumed that lower night temperatures and
higher day temperatures are favourable for the performance
of CAM, based on the observation of effects of temperature
on the overall performance of the counteracting enzymes
of nocturnal carboxylation (PEPC) and daytime decarb-
oxylation of CAM, where lower temperatures favour the
former and higher temperatures the latter (Brandon, 1967;
Kluge et al., 1973; Neales, 1973; Medina et al., 1977; Kluge
& Ting, 1978; Buchanan-Bollig & Kluge, 1981; Buchanan-
Bollig et al., 1984; Nobel, 1988; Fetene & Lüttge, 1991;
Carter et al., 1995). However, in the tropics many CAM
species perform well under rather high and similar night and
day temperatures.
Studies on species of Clusia show that temperature effects
are often closely related to effects of irradiance (Haag-Kerwer
et al., 1992). We have already seen in section IV that, depending
on irradiance during growth, day:night temperature regimes
may strongly determine the extent to which either malate or
citrate or both are used in the CAM cycle. In C. minor it was
found that the amplitude of the day:night (D:N) temperature
difference is important in eliciting a C3 to CAM switch and
this is modulated by irradiance during growth. At low irradiance,
plants performed C3 photosynthesis when D:N temperatures
were 30 : 30, 30 : 25 and 30 : 20°C and switched to CAM at
30 : 15°C. A smaller D:N temperature difference was required
for the switch in high-irradiance-grown plants which performed
C3 photosynthesis at 30 : 30°C but already switched to CAM
at 30 : 25°C. At 15 : 15°C gas exchange was reduced under
both irradiances, but high-irradiance-grown plants performed
CAM and low-irradiance-grown plants C3 photosynthesis.
The changes in the mode of photosynthesis were always readily
reversible when temperature regimes were changed again.
VI. Phenotypic plasticity: physiotypes and
morphotypes
Clusia species are woody shrubs and trees with a dichasial
cyme. In an earlier review (Lüttge, 1999) I proposed
considering all species of Clusia to belong to a single
morphotype. Saying this, I referred to the vegetative
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photosynthetic organs, the leaves, which, although they
may have greatly differing sizes in different species, are all
somewhat leathery and succulent. Diversity is revealed
when we consider the physiotypes with high plasticity of
modes of photosynthesis and metabolic pathways during
CAM performance, as described in sections III and IV. At the
morphotypic level diversity is very high in the generative
organs (Engler, 1925; Bittrich & Amaral, 1996; Gustafsson
& Bittrich, 2003; Gustafsson et al., 2006). Flower size may
range in diameter from 5 to 150 mm. There is substantial
diversity in floral morphology with respect to size, degree of
fusion and number of floral parts, position of staminodes and
stamen and anther morphology. Pollination rewards are
nectar, pollen and resin. Floral resin is a rare award. It has
evolved polyphyletically in the genus of Clusia (Gustafsson
et al., 2006). Diversity at the morphological level is also
revealed by the different life forms of Clusia species. They may
germinate terrestrially and directly develop into independent
free-standing trees. They may also germinate epiphytically,
predominantly in humus niches between branches of host
trees or of other epiphytes such as epiphyte nests or even the
tanks of bromeliads. Then they can establish soil contact via
aerial roots, i.e. they are hemi-epiphytes. Other adventitious
roots may strangle the bark of the host tree and thus murder
the phorophyte, the stem of which may then rot away under a
network of strangler roots so that the previous hemi-epiphyte
eventually becomes a free-standing tree (Lüttge, 1991). This
diversity does not only occur among species, because even
individual species can express many of these life forms, so that
we have here another example of plasticity in the genus
(Fig. 4).
In contrast to other hemi-epiphytes, mainly in the genus
Ficus, several morphological/anatomical traits, such as leaf
thickness, and stomatal density and size, did not differ much
between the epiphytic and rooted tree life form of C. minor
(Holbrook & Putz, 1996). Often the hydraulic architecture
must be adapted in hemi-epiphytes as compared with rooted
trees for sufficient water supply. In a comparative study of the
hemi-epiphytic C3/CAM-intermediate Clusia uvitana Pittier
with other hemi-epiphytes, mainly of the genus Ficus (Zotz
& Winter, 1994a,b; Patiño et al., 1995; Zotz et al., 1997), it
turned out that C. uvitana had less effective unit leaf area, a
rather large leaf area per unit of stem cross-sectional area and
a lower specific stem conductance. This suggests that physio-
typic flexibility with the water-saving CAM option also relates
to a morphotypic trait such as hydraulic architecture.
VII. Ecological amplitude and habitat impact
Physiological ecology has always largely been autecology. We
have noticed, however, that it is increasingly developing the
potential to make contributions to synecology (Lüttge &
Scarano, 2004). This is in the cast of mind of Sir Arthur
Tansley, whose name is heading the series of reviews to which

Fig. 4 Life forms of Clusia. Top row
(left–right): terrestrial seedlings of Clusia
multiflora; terrestrial seedlings of Clusia
minor; seedling of Clusia sp. under the bark
of a fallen tree. Centre row (left–right): Clusia
rosea in an epiphyte nest; roots of C. rosea in
the tank of the bromeliad Aechmea lingulata,
where the tank was cut open for
photography; C. rosea growing epiphytically
in a tree branch. Bottom row (left–right): two
pictures of C. rosea as a strangler and C. rosea
as a free-standing tree.
the current contribution belongs, because quite early ‘He
made it clear that synecology and autecology are subsumed in
the study of the community by methods firstly descriptive
and appreciative, and secondly, analytic and experimental’
(Godwin, 1977). Thus, in this vein, in this section I shall
describe the community performance of Clusia. However, one
may bear in mind here that laboratory studies of autecology
mostly cover the dynamics of short-term acclimations, while
field observations represent momentary pictures of long-term
adaptations. Long-term laboratory observations are as rare as
recordings of gradual responses under gradually varying, for
example seasonally, natural conditions. The continuous long-
term studies in the field which I allude to below are quite rare.
With the great plasticity found in the genus Clusia we may
expect that it will span a large ecological amplitude. Indeed,
it is observed that the genus, with its single simple vegetative
© The Authors (2006). Journal compilation © New Phytologist (2006) www.newphytologist.org
Tansley review Review 17
morphotype, occupies a wide range of habitats in the neotropics,
as listed in Table 2 and illustrated in Fig. 5. Not only the
genus as a whole but also some individual species, for example
the C3 species C. multiflora, the CAM species C. rosea and the
C3/CAM-intermediate species C. minor, cover almost the
entire range of different habitats listed in Table 2, and some
species occur in rather contrasting habitats, such as the coastal
restingas of Brazil and inselbergs (the C3 species Clusia parvi-
flora Saldanha et Engl.) and the restingas and dry lowland
forest (the CAM species Clusia fluminensis Pl. et Tr.). In the
following I shall list a number of sites occupied by Clusia
species and try to evaluate the environmental impact of the genus.
I shall do this by comparing the ecophysiological performance
of Clusia species with those of other co-occurring plants with
similar life forms, i.e. shrubs and trees. Much field work is
in fact reported in the literature and an ecophysiological
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18 Review Tansley review

Table 2 Ecological amplitude of Clusia with the range of habitats where Clusia is found, and occupation of a large number of these habitats
by three selected species with obligate C3 photosynthesis, obligate crassulacean acid metabolism (CAM) and C3/CAM-intermediate behaviour

Habitat C. muliflora (C3) C. rosea (CAM) C. minor (C3/CAM)

Restinga
Coastal rocks

Savanna/cerrado


Gallery forest – cerrado ecotone
Open shrubland

Dry lowland forest


Secondary shrub forest


Dry montane karstic limestone forest



Montane rainforest



Upper montane rainforest

Cloud forest/fog forest/elfin forest

Inselberg

comparison can be based on traits such as carbon isotope
ratios, rates of CO2 exchange, Chl a fluorescence parameters
of photosytem II and day:night changes of organic acid levels
in CAM-performing species. Space does not allow me to go
into as much detail as elsewhere ( Lüttge, 1999, 2006b), but
the advantage of a briefer treatment is that it provides a more
concise overview.
1. The Brazilian restinga complex
The restinga complex consists of a mosaic of plant
communities on the sandy coastal plains ranging from open
formations to forest ecosystems ( Lacerda et al., 1993). The
open formation is characterized by shrub islands surrounded
by white sand. There are sand dune ridges with dry forest in
between and on fixed dunes (Cirne & Scarano, 2001). Species
of Clusia are important floristic elements of the restingas,
which also have been called ‘Clusia scrub’ (Ule, 1901).
According to annual rainfall, height of the ground water table
and salinity of the ground water, we may distinguish wet,
intermediate and dry restingas. The major Clusia species of
the restingas are the C3 species C. parviflora, and the CAM
species C. hilariana and C. fluminensis, the former of which is
much more dominant than the latter.
In the wet restinga of Jacarepiá (22°47′ to 22°57 ′ S, 42°20′
to 42°43′ W), C. fluminensis has been compared with the C3
Fabaceae species Andira legalis Vell. Toledo (Geßler et al.,
2005a; Scarano et al., 2005). C. fluminensis developed some-
what larger maximum apparent electron transport rates of
photosynthesis (ETRmax) and with respect to photoinhibition
the two species were very similarly affected at midday and
slightly before dawn. In the intermediate restinga of Jurubatiba
(22°00′ to 22°23′ S, 41°15′ to 41°45′ W), C. parviflora and
C. hilariana were compared with the C3 species A. legalis,
Rheedia brasiliensis (Mart.) Pl. et Tr. (another Clusiaceae) and
Myrsine parvifolia A. DC. (Myrsinaceae) (Franco et al., 1996;
Duarte et al., 2005; Scarano et al., 2005). The dominant
C. hilariana (CAM) showed superior photosynthetic capacity
(ETRmax) as compared with the other four species, and the C3
species C. parviflora was similar to the other C3 species.
Chronic photoinhibition not reversible overnight was absent
or low in all species, but an advantage of CAM in avoiding
acute photoinhibition at midday was not seen. In the inter-
mediate restinga of Barra de Maricá (22°53′ S, 42°52′ W)
C. lanceolata and C. fluminensis were studied (Roberts et al.,
1996; Reinert et al., 1997). Both are potential CAM species
but δ13C values indicated that C. lanceolata was mostly
fixing CO2 by the C3 mode, while C. fluminensis was shifted
more towards CAM. Chronic photoinhibition was absent and
ETRmax and acute photoinhibition were similar in the two
species so that the relative performance of the two modes of
photosynthesis did not make much difference. In the dry
restinga of Massambaba (22°56′ S, 42°13′ W), C. fluminensis
mainly performing CAM and for comparison A. legalis,
R. brasiliensis and M. parviflora were investigated (Duarte
et al., 2005; Geßler et al., 2005a; Scarano et al., 2005). All
species showed similar photosynthetic capacity and acute
photoinhibition and there was no obvious advantage of
CAM for C. fluminensis. In the dry forest of Buziós (22°49′ S,
41°59′ W), C. fluminensis was compared with three Fabaceae
trees, Caesalpinia echinata Lam., Caesalpinia ferrea Mart. ex
Tul. and Machaerium obovatum Kuhlm. et Hoehne, and the
Euphorbiaceae Croton compressus Lam. (Geßler et al., 2005b;
Scarano et al., 2005). C. fluminensis achieved somewhat
higher ETRmax than C. compressus and higher rates than the
other species. In terms of acute and chronic photoinhibition
it was not better off than the two Caesalpinia species or
C. compressus but was superior to M. obovatum. C. fluminensis
in the very dry forest must be limited by the moisture regime
(Scarano et al., 2005) and again there appeared to be no clear
advantage of CAM at the dry site. ETRmax measurements
(µmol m−2 s−1) indicated that C. fluminensis showed a strong
dependence on a moisture gradient from wet restinga (290) to
dry restinga (160) and dry forest (100).

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 Tansley review Review 19

Fig. 5 Ecological amplitude of Clusia. Top

row (left–right): coastal restinga in Brazil with
shrubs of Clusia hilariana in the centre;
free-standing tree of C. hilariana in a
semidecidous dry forest in Brazil. Second row:
Clusia criuva on the rock outcrops of an
inselberg in Brazil; C. criuva at the ecotone of
a gallery forest with the cerrado savanna
in Brazil. Third row: four sympatric Clusia
species on karstic limestone mountains in
northern Venezuela (Clusia multiflora, Clusia
rosea, Clusia alata and Clusia sp.) and roots
of Clusia in the crevasses of the limestone
rocks. Fourth row: Unidentified Clusia species
in a tropical cloud forest, Sierra Maigualida,
southern Venezuela; C. multiflora c. 60 cm
tall in an elfin forest, Cerro Santa Ana,
Paraguana peninsula, Venezuela.

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20 Review Tansley review

Overall, the potential of Clusia species for CAM performance

may have some subtle benefits but does not appear to provide
a very obvious conspicuous advantage as compared with C3
species of similar life form. I have alluded to the fact that
Clusia may dominate the restingas in places. Moreover, several
studies have shown that Clusia species, especially C. hilariana,
can have important pioneer functions in the restingas, starting
vegetation islands on the bare sand plain and functioning as
nurse trees under the canopy of which other vegetation may
become established (Liebig et al., 2001; Dias et al., 2005). In
ecology as well as in evolution (Gould, 2002), subtle differences
can become decisive. However, functions other than photo-
synthetic capacity discussed here could be essential, for
example reproductive biology. However, in the restingas,
sexual reproduction of Clusia species is absent or very rare and
we must assume that all growth is clonal.

2. Northern coastal mountain ranges in Venezuela

In the northern coastal range of Venezuela near Caracas
(10°24′ N, 66°58′ W, 1500–1740 m asl) the same two Clusia
species, C. multiflora and C. minor, investigated in a phytotron
study in relation to the irradiance factor (section V.2) and
physiotypic plasticity described in section III were measured
in the field (Franco et al., 1994; Grams et al., 1997). In
comparison to the behaviour discussed in section V.2 the field
performance was similar (Fig. 6). The C3 species C. multiflora
potentially could achieve higher maximum rates of CO2
uptake, J CO2-max, in the light. Notwithstanding the higher
J CO -max in C. multiflora at higher irradiance, there was a
2
pronounced midday depression. Chronic photoinhibition at
dawn and acute photoinhibition at midday (Fv/Fm values Fig. 6 Photosynthetic performance of Clusia multiflora H.B.K
< 0.8) were similar in the two species. Nocturnal CO2 uptake and Clusia minor L. in the field with different daily irradiances as
in C. minor when performing CAM increased from low to indicated. JCO -max, maximum rates of CO2 uptake (positive values)
2
and CO2 release (negative values) observed in the light period (open
medium irradiance and then decreased again at the highest bars) and in the dark period (closed bars); JCO -integr, net CO2
irradiance, and this was reflected in the total integrated CO2 2
uptake integrated over 24 h; WUE, water use efficiency integrated
uptake for the dark periods and night:day oscillations of over 24 h; Fv/Fm, potential quantum yield of chlorophyll a of
organic acids (not shown). Integrated CO2 uptake over 24 h photosytem II, predawn (dark bars) and at midday (open bars). (After
was much higher under high than under low irradiance in data of Franco et al., 1994; Grams et al., 1997; Lüttge, 1999.)
C. multiflora, and in C. minor at the highest irradiance day-
time CO2 uptake was also somewhat higher and night-time
CO2 uptake was reduced as compared with the shaded plants concentrations built up during acid remobilization in the light
at the lowest irradiance (Franco et al., 1994; Grams et al., period. All four species showed slight chronic and pronounced
1997). C. minor when performing CAM always had much acute photoinhibition at dawn and midday, respectively.
higher WUE than C. multiflora.
In the Sierra de San Luis in the state Falcón (11°18′ N,
3. Semi-evergreen moist tropical forest, Barro Colorado
69°45′ W, 1200 m asl) on a karstic limestone ridge the
Island, Panama
performance of the C3 species C. multiflora was compared with
that of three CAM species, C. rosea, C. alata and an unidentified In this forest (9°10′ N, 79°51′ W) the hemi-epiphytic
species (Popp et al., 1987; Franco et al., 1994; Haag-Kerwer C. uvitana was compared with other hemi-epiphytes by Zotz
et al., 1996). The CAM species made considerable use of & Winter (1994a,b). These authors made an important
their CAM option at this site. Photosynthetic capacity was contribution to the understanding of a possible advantage
similar in all four species. However, ETRmax values were of the CAM option in C3/CAM-intermediate species. The
higher in the CAM species as a result of the high internal CO2 advantage of the CAM option in structuring hydraulic

New Phytologist (2006) 171: 7–25 www.newphytologist.org © The Authors (2006). Journal compilation © New Phytologist (2006)

architecture in hemi-epiphytes has been mentioned in section
VI. However, the two authors also covered an entire 12-
month period with numerous measurements, one of the rare
studies meeting the need for continuous long-term studies in
the field, and thus revealing an important seasonal effect. In
the wet season, with respect to both CO2 uptake integrated
over 24 h and maximum rates of J CO2, the performance of
C. uvitana was close to that of a C3 fern Polypodium crassifolium
L. and inferior to the that of the C3 orchid Catasetum
viridiflavum Hook. However, in the dry season, when C. uvitana
switched to CAM it was superior to the other two species.
4. Tropical forest on the island of Trinidad
A similar advantage of the CAM option in response to
fluctuations of habitat conditions in wet and dry seasons
was also shown for Clusia species in Trinidad: C. minor,
C. tocuchensis, C. aripoensis and C. intertexta. This is another
example of rather continuous long-term studies, where
especially for C. minor the advantage of C3/CAM switches for
performance over the seasons was demonstrated (Borland
et al., 1992, 1993, 1994, 1996; Roberts et al., 1998).
5. Inselberg, Pão de Açúcar, Rio de Janeiro
On this inselberg (22°57′ S, 43°59′ W) the C3 species
C. parviflora and the two Euphorbiaceae C. compressus and
Styllingia dichotoma Muell. Arg. form bushes and shrubbery.
The performances of all three species are quite similar (Duarte
et al., 2005; Scarano et al., 2005).
VIII. Conclusions and outlook
Clusia, the only dicotyledonous tree genus with CAM,
presents all facets of CAM and with its extraordinary plasticity
it allows the study of CAM in all its ramifications. It suggests
Fig. 7 The Atlantic rain forest (AF) of Brazil
with its peripheral ecosystems of dry forest
(DF), high-altitude rock outcrops (HA),
inselbergs (I), restinga (R) and swamp forest
(SF), all occurring within a distance of only
60–80 km from the sea (S). (Modified from
Scarano, 2002.)
© The Authors (2006). Journal compilation © New Phytologist (2006) www.newphytologist.org
Tansley review Review 21
that not so much CAM per se but the high flexibility in carbon
acquisition under variable environmental conditions in the
tropics inherent in CAM (Fig. 1) is its real ecophysiological
advantage. Plasticity with segregation and separation of
ecotypes based on the high ecological amplitude might relate
to the high speciation rate in the genus (Lüttge, 1995a,b,
2005). The radiative power of the genus is also great. This can
be seen in the Atlantic rain forest complex of Brazil, where the
Atlantic forest itself on tertiary ground is surrounded by a
variety of plant communities (Fig. 7), some of which are also
older relicts, such as the dry forest, but some of which are
quite young, such as the restingas and swamp forests on
tertiary ground. The dioecious species C. hilariana in these
communities is a migrant from the Atlantic forest with
missing or extremely limited generative propagation in the
restingas because its specific pollinators from the Euglossini
tribe, for example the bee Trigona spinipes, did not follow it to
the new habitat (Scarano et al., 2004).
My monographic review on the genus Clusia may have
shown that there is a great potential for future study of this
single diverse genus spanning all scaling levels from molecular
genetics to ecosystems.
1 While the molecular phylogeny of Clusia has advanced, its
scope and data base need to be extended and the molecular
approach must also be used in population genetics.
2 Studies of metabolism should evaluate the organization
and management of metabolic reaction modules in the flexi-
bility of photosynthetic physiotypes. Metabolomics will be
a useful approach, but some classical biochemistry is also
required, for instance regarding the plasticity in carbohydrate
metabolism and the citric acid enigma.
3 More rigorously comparative quantitative laboratory and
field studies, such as the comparison of C. multiflora and
C. minor in the phytotron and in the field discussed in this
review, should be performed to evaluate ecophysiological
capacities and potential environmental impact.
New Phytologist (2006) 171: 7–25
22 Review Tansley review
4 In addition to assessment of ecophysiology, reproductive
biology should be advanced to foster understanding of the
performance of Clusia at the community level.
Acknowledgements
I thank Dr Annie Borland, Newcastle-upon-Tyne, UK, for
valuable comments.
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