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 Blackwell Science, LtdOxford, UKBIJBiological Journal of the Linnean Society0024-4066The Linnean Society of London, 2005? 2005
841
154
Original Article

EVOLUTIONARY PROCESSES IN ORCHIDS

R. L. TREMBLAY
ET AL
.
Biological Journal of the Linnean Society, 2005, 84, 1–54. With 5 figures

Variation in sexual reproduction in orchids and its

evolutionary consequences: a spasmodic journey to
diversification
RAYMOND L. TREMBLAY1*, JAMES D. ACKERMAN2, JESS K. ZIMMERMAN3 and
RICARDO N. CALVO4†
1
Department of Biology, 100 carr. 908, University of Puerto Rico-Humacao, Humacao, Puerto Rico
00971–4300
2
Department of Biology, PO Box 23360, University of Puerto Rico-Rio Piedras, San Juan, Puerto Rico
00931–3360
3
Institute for Tropical Ecosystem Studies, University of Puerto Rico, PO Box 21910, San Juan, Puerto
Rico 00931–1910
4
Department of Biology, University of Miami, Coral Gables, FL 33124, USA

Received 8 April 2003; accepted for publication 1 April 2004

The great taxonomic diversity of the Orchidaceae is often attributed to adaptive radiation for specific pollinators
driven by selection for outcrossing. However, when one looks beyond the product to the process, the evidence for selec-
tion is less than overwhelming. We explore this problem by discussing relevant aspects of orchid biology and asking
which aspects of reproduction explain the intricate pollination mechanisms and diversification of this family. We
reaffirm that orchids are primarily pollination limited, the severity of which is affected by resource constraints. Fruit
set is higher in temperate than in tropical species, and in species which offer pollinator rewards than those that do
not. Reproductive success is skewed towards few individuals in a population and effective population sizes are often
small. Population structure, reproductive success and gene flow among populations suggest that in many situations
genetic drift may be as important as selection in fostering genetic and morphological variation in this family.
Although there is some evidence for a gradualist model of evolutionary change, we believe that the great diversity
in this family is largely a consequence of sequential and rapid interplay between drift and natural selection. © 2005
The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54.

ADDITIONAL KEYWORDS: cost of reproduction – fruit set – gene flow – genetic drift – natural selection –
Orchidaceae – pollinator limitations – resource limitation – speciation.

INTRODUCTION orchid pollination mechanisms (e.g. van der Pijl &

Dodson, 1966; van der Cingel, 1995). These reports
Early in the history of evolutionary biology, orchids
contribute to Darwin’s arguments, although it is not
had a prominent role in providing evidence for natural
often stated explicitly. Most agree that there is a link
selection. Their unusual pollination mechanisms
between orchid pollination systems and orchid diver-
attracted the attention of Darwin (1877), who argued
sity, the distinction between cause and effect is often
that they offer strong evidence both for natural selec-
not clear. In this review we argue that the predomi-
tion and for the advantages of cross-pollination. Since
nance of pollination limitation has had a significant
then, much effort has been devoted to describing
effect on the evolution of the family and propose mech-
anisms by which orchids may have diversified.
Natural selection should favour levels of reproduc-
*Corresponding author. E-mail: raymond@hpcf.upr.edu
†Current address: HDR Engineering Inc., 2202 N. West Shore tive effort that yield optimal fruit and seed set. Many
Blvd. Suite 250, Tampa, FL 33607, USA hermaphroditic plants produce far more flowers than

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54 1
2 R. L. TREMBLAY ET AL.
fruits; orchids are superlative examples of this phe-
nomenon. Low fruit-to-flower ratios in many plants
are believed to be the result of a paucity of resources
available for fruit development. This assumption
forms the basis for the resource-limitation hypothesis
whereby consistently more flowers are pollinated than
fruits are matured (Stephenson, 1981; Lee, 1988).
Regulation of maternal investment occurs through
abortion of flowers and immature fruits (Lloyd, 1980;
Stephenson, 1981) which may also be a mechanism for
regulating seed quality (e.g. Lee & Bazzaz, 1982;
Bookman, 1984; Stephenson & Winsor, 1986). The
hypothesis that resources are the ultimate limiting
factor in angiosperm reproduction has gained wide-
spread acceptance because: (a) levels of fruit matura-
tion remain unchanged following supplementary
pollination, and (b) experimental reduction of resource
availability causes elevated levels of fruit abortion
(Stephenson, 1980, 1981; Bawa & Beach, 1981; Will-
son & Burley, 1983). Thus, according to this view, vari-
ation in reproductive success should be closely tied to
the severity of resource constraints.
Flowers that fail to become fruits are not always
wasted as they may function to enhance plant fitness
through pollen donation (Willson & Rathcke, 1974).
For example, in many milkweeds (Asclepias) fruit pro-
duction is poorly correlated with the number of flowers
in an inflorescence, but the amount of pollen removed
by pollinators, an index of male fitness, is strongly cor-
related with inflorescence size (Willson & Price, 1977;
Bell, 1985; Queller, 1985). In fact, some researchers
regard the corolla as primarily a ‘male’ organ (Bell,
1985) because pollinator attractants influence fitness
through pollen donation to a much greater extent than
through seed production (Stanton, Snow & Handel,
1986). However, some evidence suggests that larger
inflorescences do not always result in proportionally
higher male fitness (Campbell, 1989).
A common thread in these arguments is Bateman’s
Principle (Bateman, 1948), which assigns the two
aspects of sexual selection (Darwin, 1871), intrasexual
competition and mate choice, to the individual sexes.
Noting the asymmetry in resource investment in off-
spring between males and females, this principle
states that: (1) for males, reproduction is limited by
access to mates, so that they must compete for oppor-
tunities to mate with females, and (2) for females,
reproduction is limited by resources and they should
therefore exercise a choice of mates to sire their rela-
tively costly offspring.
Although Bateman (1948) and others mentioned the
possibility that sexual selection operated in plants as
well as in animals, it was much later before patterns
of pollination and fruit maturation in plants were
interpreted in this context (Janzen, 1977; Charnov,
1979, 1982; Willson, 1979; Stephenson & Bertin,
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
1983). Despite the theoretical neatness and evidence
for resource constraints, variation in reproductive suc-
cess in a number of species was found instead to be
caused by low levels of pollination (Bierzychudek,
1981a; Garwood & Horvitz, 1985; Hainsworth, Wolf &
Mercier, 1985; Burd, 1994, and references therein),
from which the pollinator limitation hypothesis
emerged.
The evolutionary and ecological consequences of pol-
linator limitation are likely to differ from those of
resource limitation. If reproduction is pollen limited,
Bateman’s Principle is inapplicable (Stephenson &
Bertin, 1983). Pollen-limited female reproduction is
equivalent to saying that females are limited by access
to mates and therefore the potential for selective mate
choice is reduced under these conditions (Willson &
Burley, 1983). In fact, it would seem that any degree of
selectivity, not just that related to sexual selection, is
of dubious value when the probability of a flower
receiving pollen becomes small. While males may still
compete amongst themselves for mates under pollen-
limited reproduction (as do females), the intensity of
male-male competition is restricted (Stephenson &
Bertin, 1983; Tremblay, 1994).
The dichotomy of resource vs. pollination limitation
may be an oversimplification. Pollination limited
plants often show effects of resource constraints, so it
may be more realistic to say that such plants are
affected by both (Montalvo & Ackerman, 1987). In
fact, the theoretical model of Calvo & Horvitz (1990),
often cited as demonstrating pollination limitation,
showed that the degree of limitation is affected by the
severity of resource constraints. Furthermore, there
can be substantial variations from year to year
(Schemske & Horvitz, 1988; Vaughton, 1991).
The theoretical consequences of resource limitation,
such as female choice, are not likely to be manifested
in combination with pollination constraints. Acker-
man & Montalvo (1990) and Meléndez-Ackerman
et al. (2000) noted that some plants are pollen limited
within a season but resource limited over their life-
times. Under such conditions, the evolutionary conse-
quences would be an optimization between increased
investment in pollinator attraction (and a reduction in
allocation to ovules) and various aspects of life history,
primarily longevity, age to first reproduction and
reproductive effort within and across years (Ackerman
& Montalvo, 1990).
Orchidaceae comprise c. one fifteenth of all
angiosperms. While the intricate relationships
between orchid flowers and their pollinators have long
received a great deal of attention (Darwin, 1877; van
der Pijl & Dodson, 1966; van der Cingel, 1995), rela-
tively little consideration has been given to the fact
that orchids often exhibit low fruit-to-flower ratios
(Darwin, 1877; Ackerman, 1986a; Gill, 1989; Neiland

& Wilcock, 1998). Many studies in which researchers
have performed supplemental pollination clearly indi-
cate that these low ratios are best explained by polli-
nator limitation (see Darwin, 1877; Ackerman, 1989;
Zimmerman & Aide, 1989; Calvo & Horvitz, 1990).
Thus, orchids provide an excellent illustration of the
evolution of reproductive strategies under pollen-
limitation. In this review we discuss the ecology and
evolution of reproduction in orchids, pursuing our
principal argument that the predominance of pollina-
tion limitation explains both their intricate pollination
mechanisms as well as the diversification of the
family.
Following an overview of the essential details of
orchid reproduction, we discuss the evidence that
reproductive success (both male and female) is polli-
nation limited in orchids. We then summarize global
patterns in orchid fruit reproduction, assembling data
from almost 200 species of orchids. We then look at the
causes of pollinator limitation, resource constraints
and other ecological factors that have been shown to
limit orchid reproduction. We conclude with a discus-
sion of the relationship between variation in reproduc-
tive success, evolutionary processes, and the apparent
high rates of speciation in the Orchidaceae.
OVERVIEW OF ORCHID FLORAL BIOLOGY
Darwin (1877) produced the first treatise on orchid
pollination in order to corroborate his thesis that sex-
ual reproduction (and cross-pollination in particular)
is fundamental to organic evolution. He thoroughly
described the functional floral morphology of a num-
ber of orchids. These essays provide indirect evidence
of selection for floral characteristics that enhance the
probability of cross-pollination. Much of the subse-
quent literature has followed suit.
Three primary features, in combination, distinguish
the flowers of orchids from those of other families:
(1) The column, the fusion of male and female organs
within a single structure located at the centre of
the flower.
(2) Pollinia, tightly packed masses of pollen found in
most orchids, transported as a unit by pollinators
(Freudenstein & Rasmussen, 1997; Pacini &
Hesse, 2002); a single visit is potentially sufficient
to produce a full seed complement (e.g. Montalvo
& Ackerman, 1987; Proctor & Harder, 1994; Naz-
arov & Gerlach, 1997).
(3) Zygomorphy, whereby a labellum is often highly
modified to serve different functions (reviewed in
van der Pijl & Dodson, 1966).
The diversity of floral shapes and functional modi-
fications found across the family are largely the result
of variation in these three features.
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
EVOLUTIONARY PROCESSES IN ORCHIDS 3
Most orchids require an external pollinating agent
(Dressler, 1981). Among non-autogamous species, we
find a wide variety of pollination systems: only abiotic
and mammalian pollination are absent. Animal
groups that pollinate orchids include birds, moths,
butterflies, a wide variety of flies, numerous bees and,
to a lesser extent, wasps. Hymenopterans alone
account for the pollination of around 60% of the family
(van der Pijl & Dodson, 1966). There is also a wide
range of levels of specificity in plant–pollinator inter-
actions in the family (Tremblay, 1992). For example,
the European Herminium monorchis was visited and
presumably pollinated by 69 insect species, including
members of four different orders: Lepidoptera,
Coleoptera, Diptera and Hymenoptera (Nilsson,
1979a). Epipactis palustris has as many as 103 species
of potentially effective pollinators (Nilsson, 1978a;
Tremblay, 1992). Nevertheless, high pollinator speci-
ficity in orchid species is much more common: about
60% of orchids have only one recorded pollinator
(Tremblay, 1992). This relationship has been well doc-
umented in a number of tropical orchids that are vis-
ited by one or a few species of euglossine bee
(Ackerman, 1983; Williams & Whitten, 1983; Roubik
& Ackerman, 1987).
There are perhaps three kinds of floral rewards
among orchids. The most common type is nutritional,
consumed by the pollinators or their larvae. Nearly all
such species are nectariferous, although some produce
oils (Vogel, 1974; Steiner, 1989) and a handful offer
pollen (Gregg, 1991b; Koryan & Endress, 2001) or
pseudopollen (Dodson & Frymire, 1961; Goss, 1977;
Davies, Winters & Turner, 2000). The second type is
peculiar to orchids pollinated by male euglossine bees.
The bees are attracted by floral fragrances that they
collect for some as yet unknown aspect of mate attrac-
tion (Dressler, 1981, 1982; Williams & Whitten, 1983;
Eltz et al., 1999). A third type has not been well doc-
umented. Some Maxillaria species produce waxes and
resins (Dondon et al., 2002) that are collected by
wasps and bees (Braga, 1977; M. Whitten, pers. comm.
2002) and presumably used for nest construction.
Although most orchids offer some type of reward, an
unusually high number of species offer no reward
whatsoever. About a third of all Orchidaceae are
deceptive (van der Pijl & Dodson, 1966; Ackerman,
1986a; Nilsson, 1992). Deception may be achieved by
the flowers’ resemblance to larval food, or to the nec-
tariferous flowers of other families, or even to female
individuals of the insect pollinator (reviewed in Dafni,
1984; Ackerman, 1986a; Nilsson, 1992).
Other distinguishing features of orchid reproductive
biology include delayed ovule development (Wirth &
Withner, 1959), the large number (20 up to as many as
4 million) of dust-like seeds contained in a single cap-
sule (Arditti & Ghani, 2000), and their dependence on
4 R. L. TREMBLAY ET AL.

fungal associations for germination and seedling
establishment (Batty et al., 2001; McKendrick et al.,
2002; Rasmussen & Whigham, 2002; Selosse et al.,
2002).
POLLINATION LIMITATION IN ORCHIDS
Pollination limitation of sexual reproduction in plants
may be detected experimentally when enhanced polli-
nation elevates seed or fruit set above natural levels
(Burd, 1994). Here, we consider three aspects of pollen
limitation: (1) frequency - when pollinator abundance
is low, some flowers or individual plants may never be
visited; (2) quantity - even when pollinator visits are
frequent, the amount of pollen actually reaching the
stigmas or ovules may be low; (3) quality - the source
of pollen (whether it arrives from the same plant or a
close relative, or from an unrelated donor) can influ-
ence fruit and seed set and even the vigour of offspring
(Charlesworth & Charlesworth, 1987).
To test for pollen limitation on fruit and seed set one
must increase pollen availability to flowers in natu-
rally occurring populations. Supplemental pollination
is best done by using all the flowers on an individual.
This reveals whether or not the plant becomes
stressed by eliminating the possibility that it shunts
resources away from less intensely pollinated flowers
in favour of the experimentally pollinated ones
[Stephenson, 1981; but see Zimmerman & Pyke
(1988)].
The source of pollen used in supplemental pollina-
tion can be problematic because of the potential for
inbreeding and outbreeding depression of fruit and
seed set (Waser & Price, 1991). We distinguish
between cross- and self-pollination because rese-
archers rarely note how supplementary crossed pollen
was collected (e.g. at what distance from target
plants).
EVIDENCE FROM THE LITERATURE
Data for this analysis and that presented in subse-
quent sections were gleaned from an exhaustive
review of the literature (including Biological Abstracts
and the Science Citation Index). Data were available
for 15 species of non-autogamous orchids in which
researchers tested for pollination limitation of fruit set
by cross-pollinating all the flowers on individual
plants growing in field populations (Table 1), compar-
ing natural levels of fruit set with those obtained from
supplemental cross-pollination. As researchers rarely
stated which plants were used for pollen sources,
these data do not control for variation in pollen quality
other than that self-pollen was not used. However, the

Table 1. Natural fruit set and hand cross-pollination of non-autogamous orchids. In all studies included here, the
experiments were performed by cross-pollinating all flowers on individual plants in field populations. Differences between
natural and experimental groups are statistically significant (Wilcoxon signed rank test, tied Z-value 3.180, P = 0.002)

Natural fruit Cross-pollination

Species set (%) fruit set (%) References

Aspasia principissa Rchb. f. 8.4–10.6 61.2 Zimmerman & Aide, 1989

Calopogon tuberosus (L.) Britton, 12.4–39.9 83.0 Firmage & Cole, 1988
Sterns & Poggenb.
Calypso bulbosa var. occidentalis (L.) 11 100 Ackerman, 1981
Oakes 21–48 50–100 Alexandersson & Ågren, 1996
Cyclopogon cranichoides (Griseb.) Schltr. 26.4–61.6 96.5–97.5 Calvo, 1990a
Cypripedium acaule Ait. 0–12.9 75.0–100 Davis, 1986; Gill, 1989; Primack &
Hall, 1990; O’Connell & Johnston,
1998
Dendrobium toressae (Bailey) Dockrill 19.0 88.0 Bartareau, 1994
Encyclia cordigera (Humb., Bonpl. & 6.8 85.8 Janzen et al., 1980
Kunth) Dressler
Epidendrum ciliare L. 4.5–15.0 32.9–49.2 Ackerman & Montalvo, 1990
Isotria vertcilata Muhl. ex Willd. 10.0 25.0 Mehrhoff, 1983
Orchis boryi Rchb. f. 26.3–50.2 98.8 Gumbert & Kunze, 2001
Platanthera blephariglottis (Willd.) Lindl. 30.7–80.6 98.6 Cole & Firmage, 1984
Stelis argentata Lindl. 2.2 29.2 Christensen, 1992
Tipularia discolor (Pursh) Nutt. 18.0–25.0 47.0–89.0 Snow & Whigham, 1989
Tolumnia variegata (Sw.) Braem 4.0 88.0 Ackerman & Montero Oliver, 1985
Mean (SE) 23.1 (6.6) 71.2 (8.5)

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
 EVOLUTIONARY PROCESSES IN ORCHIDS 5

Table 2. Additional studies of non-autogamous orchids that suggest pollen limitation of fruit set. These studies include
those in which not all flowers on an individual were experimentally pollinated or in which hand-pollination was not
performed on field populations

Species Reference

Aspidogyne argentea (Vell.) Garay Singer & Sazima, 2001b

Aspidogyne longicornu (Cogn.) Garay Singer & Sazima, 2001b
Aerangis ellisii (Rchb. f.) Schltr. Nilsson & Rabakonandrianina, 1988
Brassavola nodosa (L.) Lindl. Schemske, 1980
Catasetum viridiflavum Hook. Zimmerman, Roubik & Ackerman,1989
Cleistes divaricata (L.) Ames Gregg, 1989
Comparettia falcata Poepp. & Endl. Rodríguez-Robles, Meléndez & Ackerman, 1992
Dactylorhiza sambucina (L.) Soó Nilsson, 1980
D. incarnata (L.) Soó M. T. Kuitunen, pers. comm.
Disa uniflora Berg Johnson & Bond, 1992
Diuris maculata R. Br. Beardsell et al., 1986
Goodyera oblongifolia Raf. Ackerman, 1975; Kallunki, 1976
Ionopsis utricularioides (Sw.) Lindl. Montalvo & Ackerman, 1987
Isotria verticillata Muhl. ex Willd. Mehrhoff, 1983
Liparis lilifolia (L.) Rich. ex Lindl. Whigham & O’Neil, 1991
Listera cordata (L.) R. Br. Ackerman & Mesler, 1979
L. ovata (L.) R. Br. Nilsson, 1981
Malaxis massonii Ridl. Aragón & Ackerman, 2001
Myrosmodes cochleare Garay Berry & Calvo, 1991
Orchis collina Sol. ex Russ Dafni & Ivri, 1979
O. coriophora L. Dafni & Ivri, 1979
O. laxifolia Lam. A. Fritz, pers. comm.
O. mascula L. Nilsson, 1983b
O. morio L. Nilsson, 1984
O. spectabilis (L.) Raf. Dieringer, 1982
O. spitzelli Saut. ex Koch Fritz, 1990
Paphiopedilum virens (Rchb. f.) Pfitz. Atwood, 1985
P. volonteanum (Sand.) Stein Atwood, 1985
Platanthera bifolia (L.) Rich. Nilsson, 1983a
P. chlorantha (Custer) Rchb. Nilsson, 1983a
P. ciliaris (L.) Lindl. Robertson & Wyatt, 1990
P. okuboi Makino Inoue, 1985
P. stricta Lindl. Patt et al., 1989
Prescottia densiflora Lindl. Singer & Sazima, 2001a
P. plantaginea Lindl. Singer & Sazima, 2001a
P. stachyodes Lindl. Singer & Sazima, 2001a
Pogonia japonica Rchb. f. Matsui, Ushimaru & Fujita, 2001
Prosthechea cochleata (L.) W. E. Higgins J. K. Zimmerman & J. D. Ackerman, unpubl. data
Psychilis krugii (Bello) Sauleda Ackerman, 1989
Schomburgkia tibicinis Bateman Rico-Gray & Thien, 1987
Thelymitra epipactoides F. Muell. Cropper & Calder, 1990
Thelymitra antennifera (Lindl.) Hook. f. Dafni & Calder, 1987

evidence for pollination limitation of fruit set is
unequivocal. In all cases studied, orchids given sup-
plemental pollination produced higher levels of fruit
set than those pollinated naturally (Wilcoxon sign-
rank test, Z = 3.180, P = 0.002). These differences were
often quite substantial. Levels of fruit set between two
and ten times those seen with natural pollination
were common when plants were hand pollinated. In
the case of Tolumnia variegata, hand pollinations pro-
duced a 50-fold increase compared with natural polli-
nation (Ackerman & Montero Oliver, 1985; Calvo,
1993).
In studies on 42 additional orchid species (Table 2),
researchers performed experimental hand pollination

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
6 R. L. TREMBLAY ET AL.
without specifying whether: (1) it was applied to all
the flowers on an individual; (2) they had used plants
not growing under normal field conditions, or (3) they
had used pollen from the same plant. While these
studies are not unequivocal demonstrations of polli-
nation limitation, all but one indicate that fruit set in
these orchids may be pollen-limited in the field. The
single exception is Inoue’s (1985) study of Platanthera
mandarinorum spp. hachijoensis, in which natural
levels of fruit set often approached 100% and could
not therefore be raised through supplemental
pollination.
While representing only a tiny proportion of the
20 000 or so species, these results were obtained from
taxa across the taxonomic spectrum of the family and
strongly suggest that pollination limitation of fruit set
is a common characteristic of non-autogamous
orchids.
GLOBAL PATTERNS OF FRUIT SET
In addition to summarizing data on pollination limi-
tation, we compiled a larger data set on patterns of
fruit set (Table 3). This was done in order to compare
levels of fruit set among temperate and tropical local-
ities, compare different pollinator types, explore the
contrasts between deceptive and rewarding plants and
to contrast different inflorescence sizes. The global
patterns of fruit set provide a framework for discus-
sion of the ecological and evolutionary implications of
orchid reproduction.
Data on natural levels of fruit set and results of
hand pollination were compiled from the literature for
216 non-autogamous species (92 genera). These spe-
cies are representative of the diversity of geographical
distribution, habitat and pollination systems in the
Orchidaceae. There are species from all continents
where orchids are known, including 123 temperate
and 93 tropical species. Both terrestrial and epiphytic
species, as well as rewarding (N = 84) and deceptive
(N = 132) species were considered. Most pollinator
groups are also included.
In compiling the data, we had to accommodate dif-
ferences among studies in scope, sample size, number
of populations and years, and ways of reporting fruit
set. For each species, one value of natural and exper-
imental fruit set was sought. For studies that included
more than one population and/or year, and for species
that have been studied by more than one author, the
median of all reported fruit set values was used. Fruit
set was reported either as the ratio between the total
number of fruit and the total number of flowers in the
sample or as the average of all individuals in the sam-
ple. For hand pollination, the largest value reported
was included in order to denote the highest fruit set
achievable.
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
Because data on inflorescence size are not consis-
tently reported, we assigned each species a qualitative
index according to the range of sizes most frequently
represented. Species that had an average of less than
ten flowers per inflorescence were assigned a value of
0, and species with more than ten flowers had a value
of 1. In this paper, inflorescence size is broadly defined
as the total number of flowers produced throughout
the reproductive season. To facilitate comparison, pol-
linators were grouped in broad categories: bees,
wasps, moths, butterflies, ants, flies, birds, beetles and
generalist.
A three-way non-parametric ANOVA with interac-
tion (Iman, 1974; Zar, 1996) was used to determine
whether latitude (tropical and temperate), presence or
absence of pollinator reward, and inflorescence size
(less than vs. more than ten flowers/inflorescence),
had a significant effect on median fruit set. Values
were ranked before analysis.
Pollinator reward and latitude had a significant
effect on median fruit set (Table 4). Median fruit set in
temperate species (34.6 ± 2.3; N = 123) is more than
twice that of tropical species (17.0 ± 2.1; N = 91);
deceptively pollinated species have a per cent fruit set
(20.7 ± 1.7; N = 130) half that of rewarding species
(37.1 ± 3.2; N = 84). Inflorescence size and none of the
interactions were significant (Table 4). In general,
most species have low fruit set (Fig. 1).
FRUITING FAILURE OF INDIVIDUAL PLANTS
Species vary in the percentage of flowering plants that
fail to set fruit (Table 3). Sample size for this analysis
is limited because few surveys have reported fruiting
failure (temperate N = 18, tropical N = 18; deceptive
N = 28, rewarding N = 8). A two-way non-parametric
ANOVA with interaction of geographical area and pol-
lination mechanism was performed on the ranked
80
70
60
50
Frequency
40
30
20
10
0
0 20 40 60 80 100
Per cent fruit set
Figure 1. Frequency distribution of median fruit set of
216 orchids using data from Table 3.
 Table 3. Reproductive success of non-autogamous orchids. PR, per cent pollinaria removed. PFS, per cent fruit set (open and hand pollination). IS, inflorescence
size: 0, less than ten flowers; 1, more than ten flowers. FF, per cent number of plants which do not set fruits (fruiting failure). PG, pollinator groups: 0, moths; 1,
bees; 2, wasps; 3, flies; 4, generalist; 5, butterflies; 6, ants; 7, birds; 8, beetles.

PFS

Species PR open hand IS FF PG References

TEMPERATE , DECEPTIVE
Anacamptis pyramidalis (L.) Rich. – 35.6 – 1 – 1 Neiland & Wilcock, 1998
Anoectochilus formosanus Hayata – – 86.7 – – – Shiau et al., 2002
Aplectrum hyemale (Mulh. ex Willd.) Torr. – 56.9 86.7 0 – 1 Hogan, 1983
Arethusa bulbosa L. – 15.4 – 0 84.6 1 Thien & Marcks, 1972
Bletilla striata (Thunb.) Rchb. f. – 23.8 – 0 20.0 4 Sugiura, 1995
Caladenia tentaculata Schltdl. 40.0 29.0 100.0 0 – 2 Peakall & Beattie, 1996
Calopogon tuberosus (L.) Britton, Sterns – 19.3 88.0 0 – 1 Firmage & Cole, 1988; Thien & Marcks, 1972
& Poggenb.
Calypso bulbosa (L.) Oakes var. americana – 1.0 100.0 0 99.0 1 Boyden, 1982
(R. Br.) Luer
Calypso bulbosa (L.) Oakes var. occidentalis 27.0 11.0 100.0 0 89.0 1 Ackerman, 1981
(Holz.) Calder & Taylor
Cephalanthera longifolia (L.) Fritsch – 30.2 – 1 – 1 Dafni & Ivri, 1981b
Cephalantera rubra (L.) Rich. – 5.5 – 0 – 1 Nilsson, 1983c
Ceratandra grandiflora Lindl. – 55.6 97.7 1 – 8 Steiner, 1998
Chloraea lamellate Lindl. – 15.6 76.0 1 – 4 Lehnebach & Riveros, 2003
Cleistes divaricata (L.) Ames – 57.9 95.0 0 40.0 1 Gregg, 1989, 1991a, b
Corysanthes triloba Hook. f. – 2.5 – 0 – 0 Fitzgerald, (cited in Darwin, 1877)
Cyclopogon cranichoides (Griseb.) Schltr. – 39.0 – 0 – – Calvo, 1990b
Cypripedium acaule Ait. 13.1 5.4 100.0 0 96.8 1 Primack & Hall, 1990; Gill, 1989;
Davis, 1986; O’Connell & Johnston, 1998
Cypripedium calceolus L. – 10.5 – 0 93.0 1 Nilsson, 1979b; Kull & Kull, 1991; Kull, 1998
– 33.0 96.0 0 – 1 Blivona, 2002

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
Cypripedium californicum A. Gray – 76.0 – 0 7.2 1 Kipping, 1971
Cypripedium candidum Muhl. ex Willd. – 11.6 – 0 – – Carroll, Miller & Whitson, 1984;
Curtis, 1954
Cypripedium fasciculatum Kellogg ex S. – 47.0 – 0 – 2 Kipping, 1971
Watson – 28.9 63.2 0 – 2 Lipow, Bernhardt & Vance, 2002
Cypripedium macranthos Sw. var.
rebunense (Kudo) Miyabe & Kudo
Site 1 – 8.3 25.0 0 91.7 1
EVOLUTIONARY PROCESSES IN ORCHIDS

Site 2 – 1.2 – 0 98.8 1 Sugiura et al., 2001

7
 8

Table 3. Continued

PFS

Species PR open hand IS FF PG References

Cypripedium montanum Douglas – 62.5 – 0 – 1 Coleman, 1995

ex Lindl.
Cypripedium reginae Walter – 15.9 100.0 0 – 3 Proctor, 1998
– 29.0 – 0 – 3 K. B. Gregg, unpubl. data
R. L. TREMBLAY ET AL.

Dactylorhiza fuschii (Druce) Verm. – 39.1 – 0 – 1 Dafni & Woodell, 1986; Neiland & Wilcock,
1998; Waite, Hopkins & Hitchings, 1991
Dactylorhiza incarnata (L.) Soó – 42.0 – 1 – 1 Lammi & Kuitunen, 1995
Mattila & Kuitunen, 2000
Dactylorhiza lapponica
(Laest. ex Hartman) Soó – 16.3 – 1 – 1 Neiland & Wilcock, 1998
Dactylorhiza maculata (L.) Soó – 34.5 – 1 – 1 Neiland & Wilcock, 1998
Dactylorhiza purpurella (T. & T. A. – 32.6 – – – 1 Neiland & Wilcock, 1998
Stephenson) Soó
Dactylorhiza sambucina (L.) Soó – 8.0 100.0 1 – 1 Nilsson, 1980; Pettersson & Nilsson, 1983
Disa atricapilla (Thunb.) Sw. – 64.7 – 0 – 2 Steiner, Whitehead & Johnson, 1994
Disa bivalvata (L. f.) Durand & Schinzl. – 56.5 – 0 – 2 Steiner, Whitehead & Johnson, 1994
Disa ferruginea (Thunb.) Sw. – 69.5 – 0 – 5 Johnson, 1994
Disa grandiflora L. – 6.4 – 0 – – Darwin, 1877
Disa racemosa L. f. 23.0 30.3 100.0 1 – 1 Johnson et al., 1998
Disa tenuifolia (Thunb.) Linder 39.6 43.3 81.1 0 – 1 Johnson & Steiner, 1994
Disa venosa Sw. 49.1 13.6 100.0 0 – 1 Johnson et al., 1998
Diuris maculata R. Br. 36.5 19.0 100.0 1 – 1 Beardsell et al., 1986
Drakaea glyptodon Fitz – 13.9 – 0 – 2 Peakall, 1990
Galearis spectabilis (L.) Raf. – 2.8 – – Dieringer, 1982
Herschelianthes graminifolia (Spreng.) 52.3 63.0 – 0 – 1 Johnson, 1993
Durand & Schinzl.
Isotria verticillata Muhl. ex Willd. – 5.5 43.0 0 94.7 1 Mehrhoff, 1983
Leporella fimbriata (Lindl.) A. S. George – 17.8 – 0 – 6 Peakall, 1989a; Peakall, Beattie & James, 1987
Liparis lilifolia (L.) Rich. ex Lindl. – 1.6 62.8 1 – 3 Whigham & O’Neill, 1991
Ophrys aranifera Huds. – 0.03 – – – – Delphino in Darwin, 1877
Ophrys bombyliflora Link – 21.4 – 0 – 1 Neiland & Wilcock, 1998
Ophrys insectifera L. – 8.7 – 0 – 1 Neiland & Wilcock, 1998
Ophrys sphegodes Mill. – 21.1 – 0 – 1 Neiland & Wilcock, 1998
Ophrys tenthredinifera Willd. – 55.5 – 0 – 1 Neiland & Wilcock, 1998

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
 PFS

Species PR open hand IS FF PG References

Ophrys vernixia Brot. – 7.6 – 0 – 2 Neiland & Wilcock, 1998

Orchis boryi Rchb. f. 43.6 48.5 95.8 1 – 1 Gumbert & Kunze, 2001
Orchis caspia Trautv. – 48.2 – 1 – 1 Dafni, 1983
Orchis collina Sol. ex Russ – 30.0 100.0 1 – 1 Dafni & Ivri, 1979
Orchis galilaea (Bornm. & M. Schulze) – 30.0 – 1 – 1 Bino, Dafni & Meeuse, 1982
Schltr.
Orchis israelitica H. Baumann & Dafni – 38.1 – 0 – 1 Dafni & Ivri, 1981a
Orchis italica Poir. – 14.3 – 1 – 1 Neiland & Wilcock, 1998
Orchis laxiflora Lam. ssp. palustris – 23.0 100.0 1 – 1 A.-L. Fritz, pers. comm.
(Jacq.) Asch. & Graebn.
Orchis mascula L. 52.0 7.8 100.0 1 50.0 1 Nilsson 1983b, Johnson & Nilsson, 1999
Orchis militaris L. – 13.6 – 1 – 1 Farrell 1985, Sprengel (cited in Darwin, 1877)
– 21.5 – 1 – 1 Kisseleva & Timonin, 2001
Orchis morio L. – 20.2 100.0 1 55.0 1 Nilsson, 1984
Orchis pallens L. – 13.0 – 1 – 1 Vöth, 1982 (cited in van der Cingel, 1995)
Orchis papilionacea L. – 50.0 – 0 – 1 Vogel, 1972 (cited in Dafni, 1987)
Orchis purpurea Huds. – 5.5 – 0 – 1 Neiland & Wilcock, 1998
Orchis spitzelii Saut. ex Koch – 20.4 100.0 1 24.8 1 Fritz, 1990
Pogonia ophioglossoides (L.) Ker – 28.6 100.0 0 71.4 1 Boland & Scott, 1991; Proctor, 1998
Sepapias cordigera L. – 64.5 – 0 – 1 Neiland & Wilcock, 1998
Serapias parviflora Parl. – 58.6 – 0 – 1 Neiland & Wilcock, 1998
Serapias vomeracea Briq. – 63.8 – 0 – 1 Dafni, Ivri & Brantjes, 1981
Steveniella satyroides (Steven) Schltr. 69.0 – – – – Nazarov, 1995
Thelymitra epipactoides F. Muell. – 35.0 – – – 4 Cropper, Calder & Tomkinson, 1989
Thelymitra ixioides Sw. 10.8 28.0 – – – 4 Sydes & Calder, 1993
Triphora trianthophora (Sw.) Rydb. – 5.0 90.0 0 – 1 Williams, 1994
TEMPERATE , REWARD
Acianthus sinclairii Hook. f. – 81.6 – – – – Cheeseman, cited in Darwin, 1877
Arethusa bulbosa L. – 5.0 – 0 – 1 Thien & Marcks, 1972
Cremastra appendiculata D. Don – 1.7 95.5 1 – 1 Chung & Chung, 2003

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
var. variabilis Blume – 1.8 – 1 – 1 Sugiura, 1996a
Cymbidium goeringii (Rchb. f.) Rchb. f. – 0.5 – 1 – ? Chung & Chung, 2003
Dactylorhiza fuchsii (Druce) Soó – 53.7 – 1 – 1 Dafni & Woodell, 1986
Disa uniflora Berg 52.0 45.5 – 0 – 5 Johnson & Bond, 1992
Valley – 20.0 69.5 0 – 5
Gorge – 71.0 78.5 0 – 5
Epipactis consimilis D. Don – 15.5 – 1 – 3 Ivri & Dafni ,1977
Epipactis helleborine (L.) Crantz 41.0 – – 1 – 3 Piper & Waite, 1988
EVOLUTIONARY PROCESSES IN ORCHIDS
9
 10
Table 3. Continued

PFS

Species PR open hand IS FF PG References

Epipactis palustris (L.) Crantz 64.0 62.3 – 0 – 4 Nilsson, 1978a

Epipactis thunbergii A. Gray – 54.7 – 1 – 3 Sugiura, 1996b
Goodyera foliosa (Kuntze) Benth. ex 17.5 57.1 – 0 – 1 Sugiura & Yamaguchi, 1997
Hook. f. var. maximowicziana Makino
Goodyera oblongifolia Raf. – 52.6 100.0 1 0.0 1 Ackerman 1975; Kallunki, 1976
R. L. TREMBLAY ET AL.

Goodyera procera Ker-Gawl. 75.0 92 1 – 1 Wong & Sun, 1999

Goodyera repens (L.) R. Br. – 68.3 – 1 – 1 Neiland & Wilcock, 1998
Goodyera repens (L.) R. Br. var. – 49.7 – 1 – 1 Kallunki, 1981
ophioides Fernald
Goodyera tesselata Lodd. – 41.0 – 1 – 1 Kallunki, 1981
Gymnadenia conopsea (L.) R. Br. – 63.3 – 1 – – Neiland & Wilcock, 1998
Herminium monorchis (L.) R. Br. 59.0 83.0 – 1 – 4 Nilsson, 1979a
Liparis kumokiri Maek. – 11.2 – 1 – – Oh et al., 2001
Liparis makinoama Schltr. – 0.1 – 1 – – Oh et al., 2001
Liparis reflexa (R. Br.) Lindl. – 22.3 – 1 – 3 Wallace, 1974
Listera cordata (L.) R. Br. – 69.5 85.0 1 – 4 Ackerman & Mesler, 1979
– 20.7 100.0 1 – 4 Meléndez-Ackerman & Ackerman, 2001
Listera ovata (L.) R. Br. – 1.0 100.0 1 – 4 Nilsson, 1981
Microtis parviflora R. Br. 86.0 96.6 – 1 – 6 Peakall & Beattie, 1989
Monadenia ophrydea Lindl. 94.0 93.0 100.0 1 – 0 Johnson, 1995
Orchis coriophora L. – 78.0 100.0 1 – 1 Dafni & Ivri, 1979
Orchis spectabilis L. – 5.0 100.0 0 – 1 Dieringer, 1982
Oreorchis patens Lindl. 90.7 32.9 – 1 0.0 4 Sugiura, Okajima & Maeta, 1997
Platanthera bifolia (L.) Rich. 60.1 59.4 100.0 1 – 1 Nilsson, 1983a; Mattila, 2000
Platanthera blephariglottis (Willd.) – 46.8 100.0 1 2.0 0 Smith & Snow 1976; Cole & Firmage, 1984
Lindl.
Platanthera chlorantha (Cust.) Rchb. 49.3 51.8 100.0 1 – 1 Nilsson 1978b, 1983a
Platanthera ciliaris (L.) Lindl. 27.3 – 1 – 0 Smith & Snow 1976; Robertson & Wyatt, 1990;
91.0 – 1 – 0 Gregg, 1990
Platanthera integrilabia (Correll) Luer 4.9 13.6 – – – – Zettler & Fairey, 1990
Platanthera lacera (Michx.) G. Don – 70.2 91.0 1 – – Gregg, 1990
Platanthera mandarinorum Rchb. f. ssp. 81.5 90.0 – 1 – 0 Inoue, 1986b
hachijoensis (Honda) Murata
Platanthera metabifolia F. Maek. – 11.3 – 1 40.0 0 Inoue, 1986a
Platanthera obtusata (Banks ex Pursh) – 14.5 – 1 50.0 4 Thien & Utech, 1970
Lindl.

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
 PFS

Species PR open hand IS FF PG References

Platanthera okuboi Makino – 26.5 – 1 – 0 Inoue, 1985

Platanthera stricta Lindl. 67.5 52.0 98.0 1 – 1 Patt et al., 1989
Pogonia japonica Rchb. f. 17.5 20.2 75.0 0 – 1 Matsui, Ushimaru & Fujita, 2001
11.7 5.0 – 0 – 1 Ushimaru & Nakata, 2001
Pogonia ophioglossoides (L.) Ker-Gawl. – 55.0 100.0 0 – 1 Thien & Marcks 1972; Proctor, 1998
Prasophylum odoratum R. Br. 75.0 52.0 – 1 – 3 Bernhart & Burns-Balogh, 1986
Prasophyllum romanzoffia Cham. – >75.0 – 1 – 1 Larson & Larson, 1987
Pteroglossapsis ruwenzoriensis (Rendle) – 54.8 – 1 – 8 Singer & Cocucci, 1997a
Rolfe
Satyrium bicorne Thunb. 20.1 84.7 100.0 1 – 7 Ellis & Johnson, 1999
Satyrium coriifolium Sw. 38.0 41.8 100.0 1 – 7 Ellis & Johnson, 1999
Satyrium erectum Sw. 56.0 68.2 100.0 1 – 7 Ellis & Johnson, 1999
Spiranthes lacera (Raf.) Raf. var. lacera – 57.0 100 1 – 1 Catling, 1982
Spiranthes lucida (H. H. Eaton) Ames – 0.0 100 1 – 1 Catling, 1982
Spiranthes ochroleuca (Rydb.) Rydb. – 0.0 100 1 – 1 Catling, 1982
Spiranthes romanzoffiana Cham. – 0.0 100 1 – 1 Catling, 1982
Spiranthes vernalis Engelm. & Gray – 0.0 100 1 – 1 Catling, 1982
Tipularia discolor (Pursh) Nutt. – 23.0 91.3 1 11.0 0 Whigham & McWethy, 1980; Snow &
Whigham, 1989
TROPICAL , DECEPTIVE
Aspasia principissa Rchb. f. 58.1 9.5 60.8 0 77.3 1 Zimmerman & Aide, 1989
Bletia patula Graham 17.3 27.6 – 1 6.3 1 Ackerman 1995; J. D. Ackerman &
Carromero, unpubl. data
Brassavola nodosa (L.) Lindl. 38.3 13.2 67.0 0 82.6 0 Schemske, 1980; Murren & Ellisson 1996
Bulbophyllum involutum Borba, Semir & 39.3 3.8 50.0 1 – 3 Borba & Semir, 1998; Borba & Semir 1999b;
F. Barros Borba, Sheppard & Semir, 1999
Bulbophyllum ipanemense Hoehne 92.3 3.8 70.0 1 – 3 Borba & Semir, 1998; Borba & Semir 1999b;
Borba, Sheppard & Semir, 1999
Bulbophyllum warmingianum Cogn. – 0.0 33.0 1 – 3 Sazima, 1978
Bulbophyllum weddellii (Lindl.) Rchb. f. 51.5 4.5 20.6 1 – 3 Borba & Semir, 1998; Borba & Semir 1999b;
Borba, Sheppard & Semir, 1999

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
Cochleanthes lipscombiae (Rolfe) Gray – 15.0 – 0 83.0 1 Ackerman, 1983
Coryanthes elegantium Linden & Rchb. f. – 25.0 – 0 – 1 Dodson, 1965
Coryanthes leucocorys Rolfe – 0.0 – 0 – 1 Dodson, 1965
Coryanthes macrantha (Hook.) Hook. – 21.0 – 0 – 1 Dodson, 1965
Coryanthes rodriguesii Hoehne – 43.0 – 0 – 1 Dodson, 1965
Coryanthes trifoliata C. Schweinf. – 40.0 – 0 – 1 Dodson, 1965
Corymborkis forcipigera L. O. Williams – 9.1 – 1 – – Ackerman, 1995
Cyclopogon cranichoides (Griseb.) Schltr. – 29.5 97.5 1 38.5 – Calvo, 1990a
EVOLUTIONARY PROCESSES IN ORCHIDS

Cymbidiella flabellata (Thou.) Rolfe 52.2 5.0 – 1 – 2 Nilsson et al., 1986

11
 12

Table 3. Continued

PFS

Species PR open hand IS FF PG References

Cyrtopodium broadwayi Ames – 1.0 – 1 – 1 Quesnel et al., 1982

Dendrobium infundibulum Lindl. 5.5 6.5 – 1 91.0 1 Kjellsson, Rasmussen & Dupuy, 1985
Dendrobium monophyllum F. Muell. 79.5 6.6 – 1 10.2 1 Bartareau, 1995
Dendrobium speciosum Sm. – 10.0 – 1 – 4 Calder, Adams & Slater, 1982
R. L. TREMBLAY ET AL.

Dendrobium toressae (Bailey) Dockrill 72.0 19.0 88.0 0 37.5 1 Bartareau, 1994
Dilomilis montana (Sw.) Summerh. I. Rodríguez-Colón & J. D. Ackerman,
unpubl. data
Site 1 – 6.1 – 0 92.2 1?
Site 2 – 14.3 – 0 70.3 1?
Elleanthus cf. brenesii – 29.0 – 1 – 7 B. Grabowski, pers comm.
Encyclia cordigera (Humb., Bonpl. & – 7.0 97.0 0 78.0 1 Janzen et al., 1980
Kunth) Dressler
Epidendrum ciliare L. 21.9 7.7 49.2 0 – 0 Ackerman & Montalvo, 1990
Epidendrum exasperatum Rchb. f. – 2.0 – 1 40.0 5 Calvo, 1990b
Ionopsis utricularioides (Sw.) Lindl. – 6.1 19.1 1 – 7 Montalvo & Ackerman, 1987
Laelia speciosa (Humb., Bonpl. & – 14.9 66.7 0 – 1 Hernández-Apolinar, 1992
Kunth.) Schltr.
Lepanthes caritensis Tremblay & 4.3 0.0 – 1 100.0 3 Tremblay, 1997b; Tremblay et al., 1998
Ackerman
Lepanthes eltoroensis Stimson 19.1 1.8 – 1 88.5 3 Tremblay, 1996
Lepanthes rubripetala Stimson 11.7 5.0 – 1 75.9 3 Tremblay, 1996
Lepanthes rupestris Stimson 12.6 4.9 – 1 50.7 3 Tremblay, 1996
Lepanthes sanguinea Hook. – 5.9 – 1 71.4 3 Ackerman & Zimmerman (cited in
Christensen, 1992)
Lepanthes wendlandii Rchb. f. – 12.0 – 1 34.0 – Calvo, 1990b
Lepanthes woodburyana Stimson – 9.1 – 0 62.5 – J. D. Ackerman & J. K. Zimmerman,
unpubl. data
Malaxis massonii (Ridl.) Kuntze 14.3 2.4 29.0 1 – – Aragón & Ackerman, 2001
Mormodes tuxtlensis Salazar – 3.3 – 1 30.0 1 Sosa & Rodríguez-Angulo, 2000
Myrmecophila tibicinis (Bateman) Rolfe – 2.4 92.0 0 69.3 1 Rico-Gray & Thien, 1987
Nervilia bicarinata (Bl.) Schltr. 14.6 – – 0 – 2 Pettersson, 1989
Nervilia humilis Schltr. 21.6 – – 0 – 1 Pettersson, 1989
Nervilia shirensis (Rolfe) Schltr. – 10.0 – 0 – 2 Pettersson, 1989
Nervilia stolziana Schltr. 25.9 50.0 – 0 – 2 Pettersson, 1989

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
 PFS

Species PR open hand IS FF PG References

Oncidium altissimum (Jacq.) Sw. – 2.0 – 1 – 1 Ackerman, 1995

Oncidium ascendens Lindl.
Forest 20.0 6.8 – 1 – 1 Parra-Tabla et al., 2000
Pastoral field 6.5 3.1 – 1 – 1 Parra-Tabla et al., 2000
Oncidium stipitatum Lindl. – 1.8 – 1 – 1 J. K. Zimmerman, unpubl. data
Paphiopedilum villosum (Lindl.) Stein – 7.8 – 0 – 1 Bänziger, 1996
Polystachya concreta (Jacq.) Garay & 24.5 10.0 – 1 – 1 Goss, 1977
Sweet
Pleurothallis adamantinensis Brade – – 80.8 1 – 3 Borba, Semir & Shepherd, 2001; Borba &
Semir 2001
Pleurothallis fabiobarrosii Borba & – – 78.6 1 – 3 Borba, Semir & Shepherd, 2001; Borba &
Semir Semir 2001
Pleurothallis johannensis Barb. Rodr. – 39.0 59.0 1 – 3 Borba, Semir & Shepherd, 2001; Borba &
Semir 2001
Prosthechea cochleata (L.) W. E. – 4.6 54.5 0 76.3 1 J. D. Ackerman & J. K. Zimmerman,
Higgins unpubl. data
Psychilis krugii (Bello) Sauleda 21.3 4.0 8.0 0 90.0 – Ackerman, 1989
Stelis argentata Lindl. 58.7 2.2 29.2 1 – – Christensen, 1992
Stelis sp. 1 – 15.0 – 0 – – Christensen, 1992
Stelis sp. 2 – 8.0 – 1 – – Christensen, 1992
Stelis sp. 3 – 12.0 – 0 – 8 Christensen, 1992
Stelis sp. 4 – 2.0 – 0 – – Christensen, 1992
Tetramicra canaliculata (Aubl.) Urb. – 6.0 80.0 0 – – Pagán, Martínez & Ackerman (cited in
Ackerman, 1995)
Tolumnia variegata (Sw.) Braem 3.9 2.3 77.8 0 98.0 1 Ackerman & Montero Oliver, 1985;
Ackerman, Meléndez-Ackerman &
Salguero-Faría, 1997; Calvo, 1990b
Vanilla barbellata Rchb. f. 5.3 18.2 100.0 0 – 1 I. Panetto & J. D. Ackerman, unpubl. data;
L. R. Nielsen & J. D. Ackerman, unpubl. data
Vanilla claviculata (W. Wright) Sw. 17.9 15.0 100.0 0 – 1 I. Panetto & J. D. Ackerman, unpubl. data;
L. R. Nielsen & J. D. Ackerman, unpubl. data

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
Vanilla dilloniana Correll 5.5 14.5 100.0 0 – 1 I. Panetto & J. D. Ackerman, unpubl. data;
L. R. Nielsen & J. D. Ackerman, unpubl. data
Vanilla planifolia Andrews – 1.0 – 0 – 1 Ackerman, 1995
Vanilla poitaei Rchb. f. – 6.4 100.0 0 – 1 I. Panetto & J. D. Ackerman, unpubl. data
TROPICAL , REWARD
Aerangis ellisii (Rchb. f.) Schltr. – 22.2 50.0 0 14.0 0 Nilsson & Rabakonandrianina, 1988
Angraecum arachnites Schltr. – 41.0 – 0 – 0 Nilsson, 1985; Nilsson et al., 1985,
Aspidogyne argentea (Vell.) Garay – 11.1 86.1 1 – 5 Singer & Sazima, 2001b
EVOLUTIONARY PROCESSES IN ORCHIDS
13
 14
Table 3. Continued

PFS

Species PR open hand IS FF PG References

Aspidogyne longicornu (Cogn.) Garay – 18.0 100.0 1 0.0 1 Singer & Sazima, 2001b
Catasetum macrocarpum Rich. ex Kunth – 7.1 – 0 – 1 Carvalho & Machado, 2002
Catasetum viridiflavum Hook. – 12.0 95.8 1 71.2 1 Zimmerman, Roubik & Ackerman, 1989;
Zimmerman, 1991
Comparettia falcata Poepp. & Endl. 29.1 15.7 86.5 0 – 7 Rodríguez-Robles, Meléndez & Ackerman,
1992; Ackerman,Rodríguez-Robles & Meléndez,
R. L. TREMBLAY ET AL.

1994; Salguero-Faría & Ackerman, 1999

Cyclopogon congestus (Vell.) Hoehne – 0.0 98.0 1 – 1 Singer & Sazima, 1999
Dendrochilum longbracteatum Pfitzer – 0.02 – 1 – 3 Pedersen, 1995
Erythrodes arietina (Rchb. f. & Warm.) – 22.6 – 1 0.0 1 Singer & Sazima, 2001b
Ames
Habenaria gourlieana Lindl. 15.4 19.2 – 1 33.3 0 Singer & Coccuci, 1997b
Habenaria hieronymi Kraenzl. 67.1 81.0 – 1 8.3 0 Singer & Coccuci, 1997b
Habenaria montevidensis Spreng. 49.4 61.0 – 1 0.0 0 Singer & Coccuci, 1997b
Habenaria parviflora Lindl. – 0.0 96.7 1 – 3 Singer, 2001
Habenaria rupicola Barb. Rodr. 69.5 79.3 – 1 0.0 0 Singer & Coccuci, 1997b
Leochilus scriptus (Scheidw.) Rchb. f. – 60.0 86.5 0 – 2 Chase, 1986
Myrosmodes cochleare Garay – 21.5 45.0 1 – 2 Berry & Calvo, 1991
Mystacidium venosum Lindl. – 0.0 65.0 0 – 0 Luyt & Johnson, 2001
Notylia nemorosa Barb. Rodr. – 64.8 – 1 – 1 Singer & Koehler, 2002
Pelexia oestrifera (Rchb. f. & Warm.) – 0.0 100.0 1 – 1 Singer & Sazima, 1999
Schltr.
Pleurothallis ochreata Lindl. – 12.5 39.4 1 – 3 Borba, Semir & Shepherd, 2001; Borba &
Semir, 2001
Pleurothallis racemiflora Lindl. ex – 17.3 – 1 44.4 – Ackerman, 1995
Hook.
Pleurothallis teres Lindl. – 7.0 44.3 1 – 3 Borba, Semir & Shepherd, 2001; Borba &
Semir, 2001
Prescottia densiflora Lindl. – 52.5 100.0 1 0.0 1 Singer & Sazima, 2001a
Prescottia plantaginea Lindl. – 30.0 48.1 1 – 0 Singer & Sazima, 2001a
Prescottia stachyodes (Sw.) Lindl. – 78.6 95.9 1 0.0 0 Singer & Sazima, 2001a
Sarcoglottis faciculata (Vell.) Schltr. – 0.0 95.0 1 0.0 1 Singer & Sazima, 1999
Sauroglossum elatum Lindl. – 33.4 98.1 1 0.0 0 Singer, 2002
Stenorrhychos lanceolatus (Aubl.) L. C. 74.7 83.2 – 1 0.0 7 Singer & Sazima, 2000
Rich.
Xylobium squalens Lindl. – 27.0 90.0 0 – 1 Pintaúdi, Stort & Marin-Morales, 1990

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
 EVOLUTIONARY PROCESSES IN ORCHIDS 15

Table 4. Results of three-way non-parametric ANOVA Table 5. Median per cent fruit set and standard error (SE)
with interactions for ranked median fruit set. Data of open of naturally pollinated orchids by pollinator types. Median
pollination only, from Table 3. Distribution: temperate vs. fruit set is significantly distinct among pollinator groups
tropical. Reward: deceit or reward. Inflorescence size: Inflo- (Kruskal-Wallis H, corrected for ties = 24.82, P = 0.0004).
rescence smaller than ten flowers or equal to or larger than Beetles and ants excluded because of low sample size
ten flowers
Pollinator N Fruit set SE
Source d.f. MS F P
Bees 108 25.8 2.2
Distribution 1 61707.9 19.19 <0.0001 Birds 6 53.8 11.9
Reward 1 36862.3 11.46 0.0008 Butterflies 5 36.7 12.6
Inflorescence size 1 1367.1 0.43 0.51 Generalist 8 37.7 9.3
Distribution ¥ reward 1 8698.1 2.71 0.10 Flies 29 11.9 2.7
Distribution ¥ 1 1382.1 0.43 0.51 Moths 22 41.8 6.5
inflorescence Wasps 9 34.2 7.9
Reward ¥ inflorescence 1 20.4 0.01 0.94
Three-way interaction 1 844.7 0.20 0.61
Residual 206 3215.4
Moreover, median fruit set and per cent fruiting
failure were significantly negatively correlated
(Spearman’s rank correlation, corrected for ties,
rho = -0.655; P < 0.0001; N = 44). Thus, deceptive spe-
80
Temperate cies with low overall fruit set and small inflorescence
size are most likely to exhibit high levels of fruiting
70
failure.
Tropical
60
Per cent fruiting failure

50 POLLINATOR GROUP EFFECT

Per cent median fruit set varies according to pollinator
40
group (Kruskal–Wallis H, corrected for ties = 24.82,
30 P = 0.0004; Table 5). We tested the difference between
the median fruit set of orchids pollinated by moths,
20 bees, wasps, flies, butterflies and birds (beetles and
ants were excluded because of small sample size) as
10
well as generalist orchid species. Fruit set was lowest
00 for species pollinated by flies and highest for those pol-
linated by birds and for generalists. However, the pat-
Deceptive Reward
tern may be confused by the fact that all the generalist
Figure 2. The frequency of deceptive and rewarding species with fruit set higher than 50% are temperate,
plants that fail to set fruits. Data are from Table 3. Means nectariferous species, while of the species pollinated
+ SE; N = 44. by bees, many with fruit set below 10% are tropical.

GENERAL PATTERNS OF POLLINATOR LIMITATION

data set. No difference was detected between tropical
and temperate species; mean fruiting failure was in
the range of 50–60% of plants from both areas
(F1,32 = 0.18, P = 0.67; Fig. 2). However, failure
depended on the pollination mechanism: in deceptive
species, most plants do not set fruit (66.5%); reward-
ing species have much lower failure rates (29.1%;
F1,32 = 8.28, P = 0.007).
Per cent fruiting failure was significantly higher for
species with less than ten flowers per inflorescence
(68.0% ± 6.4; mean ± SE) than for those with more
than ten (42.8% ± 8.2; mean ± SE; Mann–Whitney U-
test, Z = 2.353; P < 0.02; N = 46).
Our data demonstrate that fruit production in non-
autogamous orchids is pollen-limited. For almost all
species where data are available for natural and hand
pollination, the minimum fruit set difference of the
latter was 10% larger (mean and SE; open pollination,
26.6 ± 1.7 (N = 210); cross hand-pollination, 80.0 ± 2.6
(N = 98); Table 3). Median natural fruit set in temper-
ate species is approximately twice that of tropical spe-
cies (mean and SE: tropical 17.0 ± 2.1 (N = 91);
temperate 34.6 ± 2.3 (N = 123). It is not evident why
the former should be more efficient at setting fruit. A
possible explanation (further research is required) is
that it may be a result of population size or population

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
16 R. L. TREMBLAY ET AL.
dispersion. Tropical species are frequently organized
in small groups of individuals separated by large dis-
tances (Ackerman, 1986b; Tremblay, 1997c). Hyper-
dispersed populations are perhaps more common
among tropical epiphytes because they are immersed
in a tangled canopy.
Data are generally lacking on population dispersion
in orchids. However, an alternative explanation for
differences among temperate and tropical orchids
might be that they are due to taxonomic differences
and phylogenetic constraints. An approach to testing
this hypothesis would be to investigate fruit set in spe-
cies of the same genus that are in both regions. If fruit
production is phylogenetically constrained among
regions, then we would expect fruit set within a genus
to be more similar than among genera.
Our results are similar to those of Neiland & Wil-
cock (1998), who observed that fruit set in temperate
species is about three times as successful as that of
tropical species (38.2% vs. 13.6%). We found just a two-
fold difference, which may simply be a consequence of
different sample sizes: Neiland & Wilcock (1998)
included 96 species while this survey has 216.
CAUSES OF POLLINATION LIMITATION
Pollinator effectiveness
Orchids are visited by a number of different insects or
birds, but not all visitors are pollinators (Ackerman &
Mesler, 1979; Nilsson, 1979a, b). The relative fre-
quency of effective visits is not often quantified but
variation in visitor performance can be substantial.
For example, the most frequent visitors to Herminium
monorchis were rarely effective pollinators and even
the best carried pollinaria only about 70% of the time
(Nilsson, 1979a).
Pollinarium removal and deposition
The frequency of effective visits, quantified as polli-
narium removals and pollinations, is often very low.
Although the data show substantial variation among
populations and species, usually just under half the
flowers fail to export their pollen (Table 3).
Pollinaria removals usually exceed pollinations, so
one can express efficiency as a ratio of the former to
the latter and this can be used as an index of efficiency.
Mean ratio is 1.7 : 1 in Tolumnia variegata (Acker-
man, Meléndez-Ackerman & Salguero-Faría, 1997),
1.35 : 1 in Comparettia falcata (Salguero-Faría & Ack-
erman, 1999), 2.5 : 1 in Epidendrum ciliare (Acker-
man & Montalvo, 1990), and 1.96 : 1 in Ionopsis
utricularioides (Montalvo & Ackerman, 1987. The
range of per cent removals to per cent fruit set is large
(0.24 : 1–26.7 : 1). The most effective systems are
those of Satyrium bicorne, Vanilla barbellata
(< 0.30 : 1), while the most inefficient are those of Ste-
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
lis argentata and Bulbophyllum ipanemense (> 24.0;
Table 3; ratios not shown). Pollinarium deposition effi-
ciency among species is approximately divided equally
between efficient (< 1 : 1) and inefficient (> 1 : 1;
Fig. 3). We expect pollinaria removals to depositions to
be greater than 1 unless the pollen masses can be bro-
ken unto subunits. For example, in Herminium
monorchis the number of removals per deposition
ranged from 0.74 to 0.94 (Nilsson, 1979a). We tested
the hypothesis that removal to fruit set ratio is more
efficient in mealy pollen orchid species and found it to
be true (Mann–Whitney U-test, Z = 5.443, P = 0.001;
Table 3). Orchids with mealy pollen have a mean ratio
of 1.1 ± 0.19 : 1 (N = 36) whereas those with hard pol-
linia have a mean of 6.97 ± 1.56 : 1 (N = 21).
Pollinator abundance and diversity
Fruit set within and among populations may be influ-
enced by pollinator activity and diversity. In several
Swedish populations of Listera ovata it varied
between 13 and 70% and was positively correlated
with visitation rates (Nilsson, 1981). Visitation fre-
quencies to two populations of the epiphytic Comp-
arettia falcata were related to the abundance of its
hummingbird pollinator (Rodríguez-Robles et al.,
1992). Ackerman et al. (1997) observed the effect of
pollinator abundance and floral fragrance on fruit set
in the deceptive orchid Tolumnia variegata and found
that the main cause of fruit production among popu-
lations could be attributed to pollinator abundance.
Moreover, flower production often needs to be synchro-
25
20
15
Frequency
10
5
0
<1 2 3 4 5 6 7 8 9 10 <20 <30
Pollinaria removal per fruit set
Figure 3. Frequency distribution of pollinaria removal to
fruit set ratio. Values below 1 represent species that set
more then one fruit per pollinarium removal (mainly
orchids with mealy pollen) while values larger than 1 rep-
resent species that remove more than one pollinarium per
fruit set.

nized with the pollinator emergence for effective fruit
production (Nilsson, 1980, 1983b; Ackerman, 1981,
1983; but see Zimmerman, Roubik & Ackerman,
1989). Another source of variation is the composition
of the pollinator fauna. Robertson & Wyatt (1990)
quantitatively studied the pollination system of Pla-
tanthera ciliaris in two disjunct populations in the
south-eastern United States. They found that both
the primary pollinator and fruit set were different
in the two populations.
Pollen quantity
What proportions of ovules are fertilized during the
development of orchid fruits? While pollinia can obvi-
ously contain many thousands of pollen grains, is the
quantity of pollen deposited more than enough to fer-
tilize all ovules? Variation is to be expected as polli-
naria contain groups of two, four, six or eight pollinia,
not all of which are necessarily deposited in a single
pollination event. Moreover, orchid pollen is not
always deposited as indivisible pollinia. Pollinia may
be ‘sectile’ or ‘mealy’, that is, soft enough to be broken
apart into chunks during pollen deposition (Dressler,
1981; see Pacini & Hesse, 2002; for a review of pollen
dispersal units in orchids).
There are indications that increasing the number of
pollinia applied to orchid stigmas causes increased
seed set in orchids. In Cattleya trianaei Linden &
Rchb. f., fruit size was found to increase with the num-
ber of pollinia applied to stigmas (Duncan & Curtis,
1943). No difference in fruit set was observed in Epi-
dendrum ciliare following hand pollination with two
or four pollinia (Ackerman & Montalvo, 1990) and in
Ionopsis utricularioides with one or two pollinia (Mon-
talvo & Ackerman, 1987). Gregg (1991a) experimen-
tally investigated the effect of variable deposition on
seed development in Cleistes divaricata. As expected,
seed production declined with reduced deposition,
while seed fertility (per cent of seeds with embryos)
was unaffected.
Pollen quality
Cross- and self-pollination: Low pollination fre-
quency and pollen quantity may limit seed production
in orchids, but in self-compatible plants poor quality
pollen resulting from self-pollination can also reduce
seed production; this is not always reflected in fruit
set. We have self- and cross-pollination data for 69 spe-
cies and in 29% of these fruit set was the same. In the
remainder, fruit set from outcross pollinations was
slightly higher (81%) than from self-pollinations (72%;
Wilcoxon sign-rank test, tied Z = 3.425, N = 52,
P = 0.0006, Table 6). In general, it appears that self-
pollination does not greatly affect fruit set in orchids
in the absence of a self-incompatibility mechanism.
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
EVOLUTIONARY PROCESSES IN ORCHIDS 17
Self-pollination has been observed to reduce seed
production in orchids. Most available data report the
percentage of mature seeds bearing embryos in fruits
from plants which have been artificially cross- and
self-pollinated. Stort & Martins (1980) measured
embryo formation in 14 non-autogamous species of
Cattleya in Brazil. Comparing fruit produced using
self-pollination and cross-pollination, they observed
an average of 15.3% of seeds with embryos in the
former and 47.5% in the latter. Although seed set from
natural pollination is often better than of hand-polli-
nation, it rarely reaches 100% (Stort, 1973; Stort &
Pavanelli, 1986; Pintaúdi, Stort & Marin-Morales,
1990).
Similar results were obtained from a compilation of
the literature involving 76 species (Table 7). Embryo
formation ranged from 24% to 99% (average 64.8%)
from cross-pollination and from zero to 99%
(average 40.8%) from self-pollination. These differ-
ences were not always apparent (e.g. Ionopsis utricu-
lariodes; Montalvo & Ackerman, 1987). Variance in
embryo formation within treatments is significantly
larger in fruit from cross-pollinated plants (equality of
variance test F71,75 = 1.987, P = 0.004). This suggests
that species that are primarily cross-pollinated are
adversely affected by inbreeding whereas others are
at least insensitive to self-pollination at embryo
formation.
Three indices of seed production have been mea-
sured for a few non-autogamous orchids in order to
compare the influence of self- and cross-pollination:
weight of mature fruits, total seed mass and seed pro-
duction. The weight of mature ovaries of selfed flowers
of Diuris maculata was 77% of that of crossed flowers
(Beardsell et al., 1986). Total seed mass of fruits from
self-pollination in Platanthera ciliaris was about 70%
of that of fruits from cross-pollination (Robertson &
Wyatt, 1990). Seed production of selfed flowers in Lep-
orella fimbriata was 76% of that of crossed flowers
(Peakall, 1989a), while in Isotria verticillata it was
approximately the same (Mehrhoff, 1983). In general,
seed production from self-pollination is less than or
equal to that of cross-pollination.
Given the apparent costs of self-pollination in most
orchids, mechanisms that promote outcrossing are to
be expected. What is unusual is that in orchids these
are often structural (movement of rostellum, move-
ment of column, stigmatic maturity) and not the result
of genetic self-incompatibility, dichogamy, or unisexu-
ality (although examples of all of these mechanisms
exist in orchids). These structural mechanisms often
involve changes in size or the movement of pollinaria
in a way that prevents the placement on the stigma for
up to several minutes of pollinator foraging, through
differential drying of opposing surfaces of structures
bearing the pollinia (Dressler, 1981; Borba & Semir,
18 R. L. TREMBLAY ET AL.

Table 6. Comparison of fruit set following artificial cross- and self-pollination. Self-incompatible plants are excluded.
Cross-pollination produced higher fruit set than self pollination. Significant differences in production were noted, although
the mean differences were not large (Wilcoxon signed rank test Z-value 3.425, P < 0.0006, N = 44, N0 = 16, N ties = 6).
Values in parentheses are number of pollinations performed

Per cent fruit set

Species Self Cross References

Aerangis ellisii (Rchb. f.) Schltr. 50.0 (26) 50.0 (26) Nilsson & Rabakonandrianina,
1988
Aspasia principissa Rchb. f. 60.0 (20) 61.0 (49) Zimmerman & Aide, 1989
Aspidogyne argentea (Vell.) Garay 47.1 (51) 86.1 (36) Singer & Sazima, 2001b
Aspidogyne longicornu (Cogn.) Garay 97.3 (37) 100.0 (30) Singer & Sazima, 2001b
Brassavola nodosa (L.) Lindl. 100 (20) 67.0 (60) Schemske, 1980
Bulbophyllum warmingianum Cogn. 25.0 (12) 33.0 (12) Sazima, 1978
Caladenia tentactulata Tate 100 (4) 100 (4) Peakall & Beattie, 1996
Catasetum viridiflavum Hook. 93.3 (15) 95.8 (71) J. K. Zimmerman, unpubl. data
Cleistes divaricata (L.) Ames 100 (20) 100 (7) Gregg, 1989
Comparettia falcata Poepp. & Endl.
Site 1 53.8 (13) 86.4 (66) Rodriguez-Robles, Meléndez &
Ackerman, 1992
Site 2 64.3 (14) 86.7 (45)
Cyclopogon congestus (Vell.) Hoenhe 88.0 (49) 98.0 (47) Singer & Sazima, 1999
Cynorkis uniflora Lindl. 100 (20) 90.0 (20) Nilsson, Rabakonandrianina &
Pettersson, 1992
Cypripedium acaule Ait. 70.0 (10) 74.7 (75) Davis, 1986
90.0 (20) 100 (20) Primack & Hall, 1990
100 (18) 100 (11) O’Connell & Johnston, 1998
Cypripedium fasciculatum Kellogg ex S. Watson 78.0 (100) 82.4 (108) Lipow, Bernhardt & Vance, 2002
Cypripedium macranthos Sw. var. rebunense 50.0 (4) 25.0 (4) Sugiura et al., 2001
(Kudo) Miyabe & Kudo
Dactylorhiza incarnata (L.) Soó 98.1 (48) 95.2 (21) M. T. Kuitunen, pers. comm.
Dactylorhiza sambucina (L.) Soó 96.7 (30) 100 (29) Nilsson, 1980
Encyclia cordigera (Humb., Bonpl. & Kunth) 69.9 (119) 91.0 (211) Janzen et al., 1980
Dressler
Epidendrum ciliare L. 95.8 (48) 93.5 (31) Ackerman & Montalvo, 1990
Epipactis consimilis Don (Druce) Soó 100 (27) 100 (27) Ivri & Dafni, 1977
Galearis spectabilis (L.) Raf. 65.4 (26) 64.6 (48) J. K. Zimmerman, unpubl. data
Goodyera procera Ker-Gawl. 92 94 Wong & Sun, 1999
Habenaria parviflora Lindl. 93.3 (30) 96.7 (30) Singer, 2001
Ionopsis utricularioides (Sw.) Lindl. 88.7 (55) 89.7 (61) Montalvo & Ackerman, 1987
Isotria medeoloides (Pursh) Raf. 82 (11) 82 (11) Vitt & Campbell, 1997
Isotria verticillata Muhl. ex Willd. 32.5 (40) 36.8 (19) Mehrhoff, 1983
Lepanthes rubripetala Stimson 0.0 (11) 30.0 (44) G. Pomales & R. L. Tremblay,
unpubl. data
Lepanthes rupestris Stimson 0.0 (25) 31.0 (78) G. Pomales & R. L. Tremblay,
unpubl. data
Lepanthes woodburyana Stimson 0.0 (50) 60.9 (92) M. Mendez & R. L. Tremblay,
unpubl. data
Liparis loeselii (L.) C. Rich. 100 (70) 94 (34) Catling, 1980
Listera cordata R. Br. 100 (14) 100 (14) Meléndez-Ackerman & Ackerman,
2001
Listera ovata (L.) R. Br. 100 (27) 100 (27) Nilsson, 1981
Myrosmodes cochleare Garay 51.0 (105) 39.7 (63) Berry & Calvo, 1991
Mystacidium venosum Lindl. 40.0 (20) 65.0 (20) Luyt & Johnson, 2001
Orchis boryi Rchb. f. 97.9 (90) 98.8 (139) Gumbert & Kunze, 2001
Orchis laxifora spp. Palustris Lam. 100 (20) 100 (20) A. Fritz, pers. comm.

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
 EVOLUTIONARY PROCESSES IN ORCHIDS 19

Table 6. Continued

Per cent fruit set

Species Self Cross References

Orchis mascula L. 100 (30) 100 (30) Nilsson, 1983b

Orchis morio L. 90.0 (10) 100 (10) Nilsson, 1984
Orchis spitzelii Sauter ex Koch 95.0 (20) 100 (20) Fritz, 1990
Pelexia oestrifera (Rchb. f. & Warm.) Schltr. 81.0 (42) 100 (34) Singer & Sazima, 1999
Platanthera bifolia (L.) Rich. 100 (27) 100 (52) Nilsson, 1983a
Platanthera blephariglottis (Willd.) Lindl. 90.9 (44) 98.6 (146) Cole & Firmage, 1984
Platanthera chlorantha (Cust.) Rchb. 100 (12) 100 (12) Nilsson, 1983a
Platanthera ciliaris (L.) Lindl.
Site 1 90.9 (33) 84.8 (33) Robertson & Wyatt, 1990
Site 2 71.9 (32) 79.4 (34)
Platanthera lacera (Michx.) G. Don 91 96 Gregg, 1990
Platanthera leucophaea (Nutt.) Lindl. 63 100 Bowles, 1985 in Gregg 1990
Pleurothallis adamantinensis Brade 4.2 80.6 Borba, Semir & Shepherd, 2001
Pleurothallis fabiobarrosii Borba & Semir 5.3 78.6 Borba, Semir & Shepherd, 2001
Pleurothallis johannensis Barb. Rodr. 12.4 59.0 Borba, Semir & Shepherd, 2001
Pleurothallis ochreata Lindl. 12.4 39.4 Borba, Semir & Shepherd, 2001
Pleurothallis teres Lindl. 12.6 44.3 Borba, Semir & Shepherd, 2001
Pogonia japonica Rchb. f. 80.0 (20) 75.0 (20) Matsui, Ushimaru & Fujita, 2001
Prescottia densiflora Lindl. 69.2 (26) 100.0 (20) Singer & Sazima, 2001a
Prescottia plantaginea Lindl. 48.0 (102) 48.1 (52) Singer & Sazima, 2001a
Prescottia stachyodes Lindl. 93.7 (223) 95.9 (218) Singer & Sazima, 2001a
Prosthechea cochleata (L.) W. Higgins
Site 1 88 (16) 88 (16) Ortiz-Barney & Ackerman, 1999
Site 2 82 (16) 82 (16)
Sacoila lanceolata (Aubl.) Garay var. 100 (6) 100 (6) Catling, 1987
lanceolata
Sarcoglottis fasciculate (Vell.) Schltr. 98.0 (43) 95.0 (42) Singer & Sazima, 1999
Sauroglossum elatum Lindl. 94.9 (158) 98.1 (162) Singer, 2002
Schomburgkia tibicinia Bateman 71.2 (59) 67.4 (46) Rico-Gray & Thien, 1987
Spiranthes lacera (Raf.) Raf. var. lacera 100 (15) 100 (17) Catling, 1982
Spiranthes lucida (H. H. Eaton) Ames 100 (12) 100 (12) Catling, 1982
Spiranthes ochroleuca (Rydb.) Rydb. 100 (20) 100 (17) Catling, 1982
Spiranthes romanzoffiana Cham. 64 (7) 100 (12) Catling, 1982
Spiranthes spiralis (L.) Chevall. 75.0 35.0 Willems & Lahtinen, 1997
Spiranthes vernalis Engelm. & Gray 100 (30) 100 (30) Catling, 1982
Stelis argentata Lindl. 1.9 (54) 29.3 (185) Christensen, 1992
Tetramicra canaliculata (Aubl.) Urb. 10-14 80 J. D. Ackerman, unpubl. data
Tipularia discolor (Pursh) Nutt. 91.3 (133) 69.1 (156) Whigham & McWethy, 1980
Mean and SE 71.6 (4.0) 80.8 (2.9)

1999a; Johnson & Edwards, 2000). This provides a
question for future research: is it the very existence of
pollinaria and column that has promoted the reliance
of orchids on structural mechanisms that promote out-
crossing?
Self-incompatibility: While most orchids appear to be
self-compatible (Table 6), self-incompatibility has
been reported in orchids of diverse lineages (Table 8).
It is common in Epidendroideae, although with such a
small sample size and the limited literature on the
subject it is premature to reach a conclusion. In tem-
perate orchids, self-incompatibility has been reported
in Galearis spectabilis (Dieringer, 1982) although a
population in Maryland has been found to be self-
compatible (J. K. Zimmerman, unpubl. data). To date,
no one has attempted to describe the self-incompati-
bility mechanism in any of these orchids.
Fruit set is often extremely low in self-incompatible
orchids. In Tolumnia variegata it is often less than 2%

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
20 R. L. TREMBLAY ET AL.

Table 7. Per cent embryos formed (EF) following pollination (self vs. outcross). Results are those of artificial pollination
(results from natural pollination in parentheses). Seeds from cross-pollinated plants have significantly more embryos
(Wilcoxon signed rank test tied Z-value 6.50, P < 0.0001, N = 72, N0 = 3, N ties = 8). The variance in embryo formation is
significantly larger in self-pollinated plants (s 2 = 905.0) than in cross-pollinated ones(s 2 = 455.5; equality of variance test
F76,75 = 1.987, P = 0.004). Autogamous species have higher (mean and SE; 68.6 ± 11.3) selfing fruit set than non-autogamous
species (38.0 ± 3.5; Mann–Whitney U with ties, Z-value 2.49, P = 0.001, N = 76). No differences in fruit set between cross-
pollinated autogamous and non-autogamous species (Mann–Whitney U with ties, Z-value 1.484, P = 0.14; mean and SE;
autogamous 77.3 ± 8.0; non-autogamous, 63.8 ± 2.6)

EF (%)

Species Autogamy Self Cross Reference

Bulbophyllum weddellii (Lindl.) Rchb. f. N 68.3 40.0 Borba, Sheppard & Semir, 1999
Bulbophyllum ipanemense Hoehne N 44.4 51.8 Borba, Sheppard & Semir, 1999
Bulbophyllum involutum Borba, Semir & N 52.7 48.1 Borba, Sheppard & Semir, 1999
F. Barros
Caladenia tentactulata Tate N 74.0 73.7 Peakall & Beattie, 1996
Catasetum viridiflavum Hook. N 67.8 98.3 J. K. Zimmerman, unpubl. data
Cattleya amethystoglossa Linden & N 5.9 70.3 Stort & Martins, 1980
Rchb. f. ex Warner
Cattleya aurantiaca (Bateman) P. N. Don Y 79.4 62.8 Stort & Martins, 1980
Cattleya bicolor Lindl. N 32.8 35.5 Stort & Martins, 1980
Cattleya dormaniana (Rchb. f.) Rchb. f. N 25.2 42.5 Stort & Martins, 1980
Cattleya elongata Barb. Rodr. N 36.3 57.6 Stort & Martins, 1980
Cattleya forbesii Lindl. N 6.4 57.0 Stort & Martins, 1980
Cattleya gaskelliana Rchb. f. N 7.7 24.5 Stort & Martins, 1980
Cattleya guttata Lindl. N 25.2 54.6 Stort & Martins, 1980
Cattleya harrisoniana Bateman ex Lindl. N 2.3 36.3 Stort & Martins, 1980
Cattleya labiata Lindl. N 9.3 30.1 Stort & Martins, 1980
Cattleya leopoldii Verschaff. ex Lem. N 20.8 48.6 Stort & Martins, 1980
Cattleya loddigesii Lindl. N 11.7 76.1 Stort & Martins, 1980
Cattleya measuresiana (Willd.) Blumensch. N 23.2 65.9 Stort & Martins, 1980
Cattleya schofeldiana Rchb. f. N 7.7 28.2 Stort & Martins, 1980
Cattleya warneri Moore N 0.0 36.7 Stort & Martins, 1980
Cleistes divaracata (L.) Ames N 64.0 89.0 Gregg, 1989
Comparettia uspida Poepp. & Endl. N 97.2 95.8 Salguero-Faría & Ackerman,
1999
Cynorchis uniflora Lindl. N 36.0 61.0 Nilsson et al., 1992
Dactylorhiza sambucina (L.) Ames N 43.0 75.0 Nilsson, 1980
Disa atricapilla (Lindl.) Bolus N 82.2 94.7 Steiner, Whitehead & Johnson,
1994
Disa ferruginea (Thunb.) Sw. N 36.4 86.4 Johnson, 1994
Diuris maculata R. Br. N 94.0 82.0
Beardsell et al., 1986
Epidendrum ciliare L. N 79.3 74.5 Ackerman & Montalvo, 1990
Epidendrum nocturnum Jacq. Y 20.9 48.7 Stort & dos Santos Pavanelli,
1986
Epidendrum rigidum Jacq. Y 51.5 -- Iannotti, Stort & Morales, 1987
Epidendrum tridens Poepp. & Endl. Y 44.2 71.5 Stort & dos Santos Pavanelli,
1986
Goodyera oblongifolia Raf. N 40.0 60.0 Kallunki, 1981
N 52.7 83.8 Ackerman, 1975
Goodyera pubescens (Willd.) R. Br. N 77.0 64.5 Kallunki, 1981
Goodyera repens (L.) R. Br. var. ophioides N 36.0 63.5 Kallunki, 1981
Fernald
Goodyera tesselata Lodd. N 88.0 79.0 Kallunki, 1981
Ionopsis utricularioides (Sw.) Lindl. N 99.0 99.0 Montalvo & Ackerman, 1987

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
 EVOLUTIONARY PROCESSES IN ORCHIDS 21

Table 7. Continued

EF (%)

Species Autogamy Self Cross Reference

Laelia caulescens Lindl. N 33.5 66.6 Stort & de Lima Galdino, 1984
Laelia cinnabarina Bateman N 59.9 70.5 Stort & de Lima Galdino, 1984
Laelia crispa Rchb. f. N 8.7 42.1 Stort & de Lima Galdino, 1984
Laelia crispilabia A. Rich. ex R. Warner N 16.6 54.0 Stort & de Lima Galdino, 1984
Laelia flava Lindl. N 12.7 39.3 Stort & de Lima Galdino, 1984
Laelia grandis Lindl. & Paxton N 9.4 41.5 Stort & de Lima Galdino, 1984
Laelia longipes Rchb. f. N 0.2 29.3 Stort & de Lima Galdino, 1984
Laelia millerii Blum N 0.0 26.6 Stort & de Lima Galdino, 1984
Laelia mixta Hoehne N 14.4 48.1 Stort & de Lima Galdino, 1984
Laelia ostermayerii Hoehne N 0.7 52.5 Stort & de Lima Galdino, 1984
Laelia perrinii Rchb. f. N 42.2 69.7 Stort & de Lima Galdino, 1984
Laelia pumila Rchb. f. N 33.0 52.6 Stort & de Lima Galdino, 1984
Laelia tenebrosa Rolfe N 12.9 46.5 Stort & de Lima Galdino, 1984
Laelia xanthyna Lindl. ex Hook. N 8.5 38.1 Stort & de Lima Galdino, 1984
Leporella fimbriata (Lindl.) A. S. George N 40.3 50 Peakall, 1989a
Listera cordata R. Br. N 88.5 94.2 Meléndez-Ackerman & Ackerman,
2001
Liparis loeselii (L.) ex Lindl. Y 95.0 95.0 Catling, 1980
Listera ovata (L.) R. Br. N 89.3 97.7 Nilsson, 1981
Oeceoclades maculata (Lindl.) Lindl. Y 92.0 88.0 González-Díaz & Ackerman, 1988
Orchis mascula L. N 59.8 75.1 Nilsson, 1983a
Orchis morio L. N 10.1 35.2 Nilsson, 1984
Orchis spitzelii Saut. ex Koch N 54.0 86.9 Fritz, 1990
Platanthera bifolia (L.) Rich. N 43.8 84.2 Nilsson, 1983b
Platanthera chlorantha (Cust.) Rchb. N 23.9 73.5 Nilsson, 1983b
Platanthera ciliaris (L.) Lindl. N 66.0 76.0 Gregg, 1990
Platanthera lacera (Michx.) G. Don N 67.0 47.0 Gregg, 1990
Pleurothallis adamantinensis Brade N 28.0 92.0 Borba, Semir & Shepherd, 2001*
Pleurothallis fabiobarrosii Borba & Semir N 30.0 94.0 Borba, Semir & Shepherd, 2001*
Pleurothallis johannensis Barb. Rodr. N 20.0 91.0 Borba, Semir & Shepherd, 2001*
Pleurothallis ochreata Lindl. N 28.0 95.0 Borba, Semir & Shepherd, 2001*
Pleurothallis teres Lindl. N 5.0 90.0 Borba, Semir & Shepherd, 2001*
Sacoila lanceolata (Aubl.) Garay var. N 80.0 75.0 Catling, 1987
lanceolata
Sacoila lanceolata (Aubl.) Garay var. Y 97.5 97.5 Catling, 1987
paludicola (Luer) Sauleda, Wunderlin
& B. F. Hansen
Satyrium bicorne Thunb. N 14.8 66.1 Ellis & Johnson, 1999
Satyrium coriifolium Sw. N 29.3 65.6 Ellis & Johnson, 1999
Satyrium erectum Sw. N 14.5 57.9 Ellis & Johnson, 1999
Sophronitis purpurata (Lindl. & Paxton)
C. Berg & M. W. Chase N 11.6 43.8 Stort & de Lima Galdino, 1984
Vanilla claviculata (W. Wright) Sw. N 63.0 88.0 L. R. Nielsen & J. D. Ackerman,
unpubl. data
Xylobium squalens Lindl. N 74.7 82.3 Pintaúdi, Stort & Marin-Morales,
1990
Mean and SE 40.8 (3.8) 64.6 (2.5)

*Values are approximate, from interpretation of figure 2 in mentioned reference

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22 R. L. TREMBLAY ET AL.

Table 8. Self-incompatible species

Species References

Acampe pachyglossa var. renschiana (Rchb. f.) Senghas Agnew, 1986

Acampe praemorsa (Roxb.) Blatt. & McCann Agnew, 1986
Angraecum cultiforme Summerh. Agnew, 1986
Cattleya warneri Moore Stort & Martin, 1980
Dendrobium (44 spp.) Johansen, 1990
Doritis pulcherrima Lindl. Agnew, 1986
Epidendrum cinnabarium Salzm. ex Lindl. East, 1940
Epipactis atrorubens (Hoffm. ex Bernh.) Besser East, 1940
Galearis spectabilis (L.) Raf. Dieringer, 1982
Malaxis massonii (Ridl.) Kuntze Aragón & Ackerman, 2001
Lepanthes woodburyana Stimson G. Pomales, M. Méndez & R. L. Tremblay, unpubl. data
Liparis lilifolia (L.) Lindl. Whigham & O’Neil, 1991
Liparis makinoana Schltr. Oh et al, 2001
Oeoniella polystachys (Thouars) Schltr. Agnew, 1986
Oncidium ascendens Lindl. Parra-Tabla et al., 2000
Oncidium cavendishianum Bateman East, 1940
Oncidium cimiciferum (Rchb. f.) Beer East, 1940
Oncidium crispum Lodd. East, 1940
Oncidium divaricatum Lindl. East, 1940
Oncidium lemonianum Lindl. East, 1940
Oncidium sphacelatum Lindl. East, 1940
Oncidium unicorne Lindl. East, 1940
Phalaenopsis schilleriana Rchb. f. Agnew, 1986
Psychilis krugii (Bello) Sauleda Ackerman, 1989
Psychilis monensis Sauleda S. Aragón, unpubl. data
Sobennikoffia humbertiana H. Perrier Agnew, 1986
Stelis argentata Lindl. Christensen, 1992
Tolumnia variegata (Sw.) Braem Ackerman & Montero Oliver, 1985
Trichocentrum microchilum (Bateman ex Lindl.) East, 1940
M. W. Chase & N. H. Williams

(Ackerman & Montero Oliver, 1985; Calvo, 1993; Ack-
erman et al., 1997) and Ackerman (1989) recorded a
value of 4% for Psychilis krugii while Christensen
(1992) reported 2.2% for Stelis argentata. J. K. Zim-
merman (unpubl. data) observed a level of 1.8% in a
Panamanian population of Oncidium stipitatum and
we observed 1.6% fruit set in a Puerto Rican popula-
tion of O. altissimum. In the tropical terrestrial Mal-
axis massonii, fruit set was 1.4% and 3.4% in two
populations (Aragón & Ackerman, 2001). Dieringer
(1982) recorded an average fruit set of 5% in two Ohio
populations of Galearis spectabilis (the self-compati-
ble Maryland population, meanwhile, exhibited fruit
set of 53%; J. K. Zimmerman, unpubl. data). Average
fruit set over 2 years in a Maryland population of
Liparis lilifolia was 1.6% (Whigham & O’Neil, 1991).
One suggestion as to why fruit set is higher in some
species is that they do not possess elaborate mecha-
nisms, evident in many other orchids (Dressler, 1981),
which prevent self-pollination. Low fruit set in self-
incompatible species may be the result of much self-
and geitonogamous pollination, resulting in flower or
fruit abortion. Alternatively, mechanisms for prevent-
ing self-pollination may include sequential flowering
(Psychilis spp. - Ackerman, 1989; Malaxis massonii -
Aragón & Ackerman, 2001), dichogamy/protandry
(Goodyera oblongifolia - Ackerman, 1975; Spiranthes
- Catling, 1983a; Prescottia stachyodes - Singer &
Sazima, 2001a; Erythrodes arietina - Singer & Saz-
ima, 2001b; Sauroglossum elatum - Singer, 2002;
Mesadenella cuspidate (Lindl.) Garay - Singer, 2002;
Notylia nemorosa Barb. Rodr. - Singer & Koehler,
2002) and temporal variation in pollinarium size (Bul-
bophyllum weddellii (Lindl.) Rchb. f., B. ipanemense
Hoehne and B. involutum - Borba & Semir, 1999a).
Autogamy: This is not infrequent among orchid spe-
cies. Van der Pijl & Dodson (1966) estimated the
family-wide occurrence of autogamy to be about 3%,
but recent estimates from local floras suggest it may

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
 EVOLUTIONARY PROCESSES IN ORCHIDS 23

be higher. Catling (1990) suggested that autogamous
pollination may occur in between 5% and 20% of the
family. Ackerman (1985) estimated that 15% of the
North American orchid flora was autogamous com-
pared to 10% for Barro Colorado Island, Panama and
25% for Puerto Rico.
There is evidence that autogamy increases with lat-
itude and in insular areas. In eastern Canada, for
example, 17% of orchids are autogamous, while values
for Europe range from 27 to 50% (Kirchner, 1922a;
Hagerup, 1952; Catling, 1983b, 1990). Self-pollination
is more frequent in austere colder habitats (Catling,
1990 and references therein) where pollinator activity
might be unpredictable.
Autogamy in orchids is often suggested by high lev-
els of fruit set. Among orchids in which mechanisms of
self-pollination are known (Table 9), average fruit set
is high, 77.0 ± 5.0% (between 14% and 100%, N = 21),
much higher than it is in allogamous orchids (Table 3).
Clearly, one result of autogamy is that pollination lim-
itation is reduced (depending on the degree to which
the pollen of a single flower can fertilize all potential
ovules) or absent and the evolution of self-fertilization
under these conditions follows the Reproductive
Assurance Hypothesis of Hagerup (1952) and Jain
(1976).
While fruit production is assured under self-fertili-
zation, there are trade-offs involved. Inbreeding
depression, the loss in offspring fitness due to the
expression of deleterious recessive alleles and other
causes (Charlesworth & Charlesworth, 1979, 1987;
Dudash, 1990), can offset increases in seed production.
However, because offspring from self-pollination pos-
sess two copies of the parental genome compared to
one in outcrossed offspring, the critical level of
inbreeding depression determining the evolution of
autogamy is assumed to be 50% (Charlesworth &
Charlesworth, 1979; Lande & Schemske, 1985; but see
Holsinger, 1988). On this basis Gill (1989, 1996) has
suggested that high inbreeding depression may
explain the failure of Cypripedium acaule to develop
autogamy from its current ‘inefficient’ mating system
(natural fruit set is often < 2% in this species, Table 3).
Data on embryo formation from cross- and self-
pollination in non-autogamous species do not suggest
a level of inbreeding depression greater than 50%

Table 9. Natural fruit set (per cent fruits from flowers) in autogamous orchids

Species % FS Location Reference

Aplectrum hyemale (Mulh. ex Willd.) Torr. 64 Illinois, USA Hogan, 1983

Bletia stenophylla Schltr. 88 Venezuela R.N. Calvo, unpubl. data
Caularthron bilamellatum (Rchb. f.) R. E. Schult. 86 Panama J.K. Zimmerman, unpubl. data
Cephalanthera austinae (A. Gray) A. Heller 14 California, USA Kipping, 1971
C. odontorhiza (Willd.) Poir. 84 Maryland, USA J.K. Zimmerman, unpubl. data
C trifida Chatel. 93 Ontario, Canada Catling, 1983b
Disa glandulosa Harv. ex Lindl. 78 South Africa Johnson, Steiner & Kurzweil,
1994
Disa vaginata Burch. ex. Lindl. 74 South Africa Johnson, Steiner & Kurzweil,
1994
Goodyera procera Ker-Gawl. 94 Hong Kong Wong & Sun, 1999
Isotria medeoloides (Pursh) Raf. 67 Main, USA Vitt & Campbell, 1997
Isotria verticillata Muhl. ex Willd. 57 Southeastern USA Mehrhoff, 1983
Liparis loeselii (L.) Rich. 71 Germany Kirchner, 1922b
Oeceoclades maculata (Lindl.) Lindl. 36 Florida, USA Calvo, 1990b
52 Puerto Rico González-Díaz & Ackerman,
1988
Ophrys apifera Huds. 100 England Darwin, 1877
Phragmipedium lindenii (Lindl.) Dressler & N. H. >95 Ecuador L. McCook, pers. comm.
Williams
Platanthera clavellata (Michx.) Luer 50 USA Gregg, 1990
Platanthera hyperborea (L.) Lindl. 99-100 Michigan, USA Catling, 1983b
Sacoila lanceolata (Aubl.) Garay var. paludicola 100 Florida, USA Catling, 1987
(Luer) Sauleda, Wunderlin & B. F. Hansen
Thelymitra carnea R. Br. 100 Australia Fitzgerald in Darwin, 1877
Thelymitra circunsepta Fitzg. 96 Australia Sydes & Calder, 1993
Mean (SE) 78.1 (4.8)

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
24 R. L. TREMBLAY ET AL.
(Tables 6, 7). The level of inbreeding depression in the
self-compatible Puerto Rican Prosthechea cochleata
showed very little difference in fitness among self and
cross progeny (Ortiz-Barney & Ackerman, 1999); in
this species a selfing mutant could easily swamp the
outcrossing phenotype. However, more thorough stud-
ies of inbreeding depression focusing on the complete
orchid life span are required to address this issue.
Comparison of seed formation in autogamous
orchids with that of outcrossing species suggests the
elimination of an inbreeding fitness bottleneck in
ancestral outcrossing stock. The autogamous Brazil-
ian Cattleya aurantiaca (Bateman) Don studied by
Stort & Martins (1980) did better under self- (79.5%)
than cross-pollination (62.8%). In Oeceoclades macu-
lata and Sacoila lanceolata (Catling, 1987; González-
Díaz & Ackerman, 1988) embryo formation was
similar in fruits from cross- and self-pollination
(Table 7). In addition, seed mass of fruits in
O. maculata was unaffected by self- and cross-
pollination (González-Díaz & Ackerman, 1988).
Facultative autogamy may occur in a number of spe-
cies (for review see Catling, 1990) and may be an
appropriate strategy when frequency of pollination is
habitually low. An occasional crossing event may be
sufficient to infuse enough genetic variability to
diminish any effects of inbreeding depression from fre-
quent self-pollination. However, supporting evidence
is not yet at hand.
Pollen diversity: Increasing pollen diversity on the
stigma can positively affect the quality and quantity of
the progeny (Marshall & Ellstrand, 1986; Montalvo,
1992). For orchids, the effects of multiple parentage
are incompletely known yet it may be important for
some species, particularly those with mealy or sectile
pollen. The incidence of multipaternal pollination in
orchids is, similarly, usually not known, although it is
not likely to be frequent, as many orchid flowers are
not visited at all and when they are, they wither after
pollen deposition (Arditti, 1976; Proctor & Harder,
1995). Nevertheless, multipaternal pollinations are
possible because pollinators may carry more than one
pollinarium (Nilsson et al., 1987; Luyt & Johnson,
2001). When evidence for such pollination has been
sought under field conditions, it was either not
detected (Tremblay, 1994; Peakall, 1989b; Nilsson,
Rabakonandrianina & Pettersson, 1992; Salguero-
Faría & Ackerman, 1999) or found to be rare (Folsom,
1994). What is the effect of pollen diversity on seed
quality? Tremblay (1994) hand-pollinated Cypripe-
dium calceolus, where treatments included pollination
by outcrossing with a single father or a mixture of ten
different fathers. Multipaternal pollination resulted
in a higher germination rate although embryo sizes
were generally smaller.
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
POLLINATION LIMITATION SUMMARY
In nearly all non-autogamous species examined, fruit
set may be increased by hand pollination, which sug-
gests that the observed variation in fruit set is mainly
dependent upon the level of pollinator activity. This in
turn may vary among populations and among years.
Thus orchids are generally pollination limited and
severely so. Pollen quantity, quality and frequency of
selfing may also affect fruit and seed set, but are gen-
erally not correlated with the degree of pollinator
activity.
RESOURCE CONSTRAINTS IN ORCHIDS
SHORT-TERM EFFECTS
What role does the availability of resources play in
orchid reproduction? The usual evidence for resource
limitation of fruit and seed set is that additional pol-
lination fails to increase fruit production, whereas an
augmentation of resources succeeds in doing so. An
experimental study evaluating the effect of nutrient
application (N, P and K) to Platanthera bifolia and
Dactylorhiza incarnata showed mixed results (Mattila
& Kuitunen, 2000). They found that for P. bifolia,
plant size was the factor in determining fruit set of fer-
tilized plants. Small plants had higher fruit set when
given nutrients, whereas large plants showed no dif-
ferences (Mattila & Kuitunen, 2000). Nutrient fertili-
zation of D. incarnata did not effect fruit initiation and
production, or leaf area in the following year (Mattila
& Kuitunen, 2000).
Another experiment evaluated the effect of addi-
tional water resources on fruit production in Platan-
thera bifolia (Mattila, 2000). While it resulted in
higher fruit production, no increase in leaf area was
noted on plants the following year (Mattila, 2000).
Fisher (1992) examined the consequences of the asso-
ciation of the self-pollinating Caulathron bilamella-
tum with an ant which inhabits the pseudobulb. The
refuse of the ant community presumably fertilizes the
plant. Such uptake has been demonstrated in Myrme-
cophila tibicinis (Rico-Gray et al., 1989). Fisher’s
removal of the ants and their refuse reduced the num-
ber of fruits produced, which suggests that reproduc-
tion is resource limited. Zimmerman & Whigham
(1992) showed that severance of old corms in Tipu-
laria discolor and experimental defoliation (herbivory
effect) had little effect on fruit production. Aragón &
Ackerman (2001) had similar results in their defolia-
tion experiments with Malaxis massonii as did Pri-
mack, Miao & Becker (1994) in their work on
Cypripedium. Not surprisingly, the response level was
dependent on the severity of the costs. Multi-year
defoliation of T. discolor resulted in failure to produce
flowers (Whigham, 1990).

Fire has been thought to stimulate flowering in ter-
restrial orchids, although Barnett (1984) found that
among 22 species at one site in Australia most were
adversely affected and only a few responded positively.
Primack et al. (1994) noted that fire also had a nega-
tive effect on Cypripedium acaule that became addi-
tive when coupled with simulated herbivory. D. E. Gill
(pers. comm.) noted a positive effect on the same spe-
cies, but an indirect one: fire opened up the forest can-
opy, stimulating flowering of food plants for the
pollinators of C. acaule, while the orchid benefited
from increased pollinator populations and visitation
rates.
Disease can affect resource availability and repro-
ductive success in many plants (Burdon, 1987), but it
rarely has been studied in natural populations of
orchids. Meléndez & Ackerman (1993) studied a com-
mon rust disease in a population of the epiphytic Tol-
umnia variegata and found that it had little impact on
either short-term vegetative growth or reproductive
success.
In some studies in which fruit production was
significantly increased by hand pollination, within-
season effects of resource availability have been
documented. These include reduced flower production
or vegetative growth, decreased fruit size with
increasing fruit number, position-dependent fruit
size, and abortion of initiated fruits (Montalvo & Ack-
erman, 1987; Ackerman, 1989; Zimmerman & Aide,
1989; Calvo, 1993). A series of experiments evaluat-
ing the effect of a reduction in photosynthesis demon-
strated that it affected neither fruit production nor
leaf area in Dactylorhiza maculata, which suggests
that orchids either are either heavily dependent on
mycorrhizae or utilize stored resources for fruit devel-
opment, capsule weight and subsequent growth (Val-
lius & Salonen, 2000; Vallius, 2001). In addition to
these short-term effects that are widely known among
orchidists, long-term costs to elevated fruit set also
occur.
LONG-TERM EFFECTS
Ample evidence for within-season pollinator limita-
tion in orchids exists, yet lifetime fruit production may
be resource-limited rather than pollinator-limited
(Montalvo & Ackerman, 1987; Zimmerman & Aide,
1989). In a study in which hand pollination resulted in
a 13-fold increase in fruit set over natural pollination
in Encyclia cordigera, Janzen et al. (1980) argued that
the results of single-season experiments could be mis-
leading because the experimental increase in fruit set
might have detrimental effects on future growth and
reproduction (i.e. it may result in an increased cost of
reproduction). Determining the long-term effect of
increasing fruit set was tested on the epiphyte, Ionop-
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
EVOLUTIONARY PROCESSES IN ORCHIDS 25
sis utricularioides, by Montalvo & Ackerman (1987).
Natural fruit set in this species was 6.1%; in hand-
pollinated plants it was between 51% and 66%, about
a 10-fold increase. However, this increase carried a
cost. There was a significant negative linear correla-
tion between fruit set in 1982 and the per cent change
in leaf area between 1982 and 1983, and plants with
>25% fruit set also had a significantly lower probabil-
ity of flowering in 1983 than those with lower fruit set
(67% failed to flower vs. 19%, respectively). Further-
more, mineral fertilization in a shaded greenhouse
resulted in higher growth rates than in the field.
These data indicate that the availability of nutrients
was a limiting factor for growth and reproduction, and
that there was a trade-off in resource allocation to
these two functions. Significant negative effects of
artificially increased fruit set on growth have been
reported in four other tropical epiphytes and two tem-
perate species (Ackerman, 1989; Snow & Whigham,
1989; Zimmerman & Aide, 1989; Ackerman & Mon-
talvo, 1990; Primack & Hall, 1990; Meléndez-
Ackerman, Ackerman & Rodríguez-Robles, 2000).
However, no effects of fruit set on either growth or
reproduction were detected in multiyear studies of
Cyclopogon cranichoides (Calvo, 1990a) and Cleistes
divaricata (Gregg, 1989).
RESOURCE CONSTRAINTS AND OVULE DEVELOPMENT
One corollary of the resource limitation hypothesis
(Stephenson & Bertin, 1983) is that the relative cost of
flowers is small compared to that of fruits, such that
large floral displays are relatively inexpensive. For
example, Bookman (1984) showed that the biomass
and nutrient content of abscised flowers and fruits of
Asclepias speciosa Torr. was < 3.2% that of mature
pods, suggesting that the cost of aborting the struc-
tures was relatively small. Similar adaptations to pol-
lination and resource limitation may have occurred in
orchids.
As noted previously, hyperovulate ovaries (and the
enormous number of seeds produced per fruit) is a
hallmark of orchid reproduction. Although first
described in the 1940s (Duncan & Curtis, 1943; see
Wirth & Withner, 1959 for citations), it is still not
widely recognized that ovule development in orchids is
usually completed after pollination. As a result, the
period between pollination and fertilization in orchids
can be quite long, ranging from a week to almost a
year (Wirth & Withner, 1959). In orchids with single
anthers, ovaries develop to a stage where three pla-
cental ridges, consisting of undifferentiated tissue, are
evident. Cypripedoid orchids have two anthers and
differentiation proceeds to where megaspore mother
cells (which will divide by mitosis to form megagame-
tophytes) are recognizable. In both types of orchids
26 R. L. TREMBLAY ET AL.
further development of ovules is triggered by auxin
released from pollen that has been deposited on the
stigma (Wirth & Withner, 1959).
Can the incomplete development of orchid ovaries
be interpreted as part of a reproductive strategy
related to pollen-limited fruit reproduction? If flowers
frequently remain unpollinated, then the arrested
development of the ovaries may represent a strategy
for limiting the cost of flower production (Dressler,
1981) in favour of pollinator attraction. Following
Bookman’s (1984) example, Zimmerman (unpub-
lished) collected data on the relative mass of flowers
and fruits from a Maryland population of Tipularia
discolor. Plants were sampled at anthesis, at the time
the ovaries of pollinated flowers had expanded (and
included only the placental ridges in the hollow
ovary), and when fruits were mature. Ovaries at
anthesis weighed about 0.5 mg (dry weight after >
72 h at 60 ∞C). The average mass of expanded ovaries
was 6.25 mg (12.5 times that of ovaries at anthesis)
and that of mature fruits was 8.65 mg (over 17 times
that of an unexpanded ovary). Thus, the mass of
mature fruits is only about 1.4 times that of mature
flowers and if T. discolor were to regularly abort
unpollinated, fully developed flowers, it would be rel-
atively costly in terms of biomass.
A test of this idea would come from an orchid in
which fruit set of flowers was assured. Wirth & With-
ner (1959) noted one exception in orchids in which
ovary development proceeds, prior to pollination, to
levels beyond those observed in other orchids. This
occurs in a triandrous form of the normally monan-
drous Prosthechea cochleata. This form is autogamous
and differentiation of the placental ridges precedes
contact of the pollen and stigmatic fluids (Duncan &
Curtis, 1943). One consequence of autogamy is that
pollen-limitation is alleviated and fruit set is assured.
In autogamous orchids, therefore, one would expect to
observe a relatively greater degree of ovule develop-
ment in advance of pollination.
EVALUATING SHORT AND LONG-TERM COSTS
OF FRUIT SET
Calvo & Horvitz (1990) investigated the relative roles
of pollination level and cost of reproduction on overall
fitness using a transition matrix demographic model
of orchid life-history. The model incorporated life-his-
tory traits characteristic of orchids, such as skewed
distribution in fruit production, the ability to regress
in size in subsequent years, and high fecundity having
a cost. Through simulation, the model predicted that
maximum fitness would be attained at intermediate
levels of pollination in most cases. The results stressed
the importance of the trade-off between the additional
fecundity gained through increased fruit production
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
and its effects on subsequent growth. For a given mag-
nitude of cost, there was a threshold return in fecun-
dity beyond which higher fruit production was
favourable.
Tolumnia variegata provides a good example of the
disparity between an increase in fruit set and the cost
of the increase. In Calvo’s (1993) study population,
hand-pollination of field plants resulted in 72% fruit
set vs. 0% in naturally pollinated plants. In the follow-
ing year the former were, on average, about 30%
smaller than the latter. This model showed that the
cost was too insignificant to override the potential ben-
efit of higher fruit set.
Re-examination of the evidence for Ionopsis utricu-
larioides (Montalvo & Ackerman, 1987) and Aspasia
principissa (Zimmerman & Aide, 1989) in terms of
their demographic model, led Calvo & Horvitz (1990)
to suggest that these species may indeed be pollinator-
limited, mainly because the fecundity gained through
higher fruit production appeared to be relatively
higher than the negative effects of costs. Meléndez-
Ackerman et al. (2000) took a different approach from
Calvo & Horvitz (1990). From experimental data on
Comparettia falcata, they calculated a relative fitness
index to compare pollen augmented plants and natu-
rally pollinated plants and found that lifetime fitness
in this species can be resource limited. Nutrient avail-
ability affects flowering in a number of orchids and
consequently resources do affect reproductive success
(Dijk, Willems & Van Andel, 1997). Nevertheless, the
available evidence seems to indicate that, although
resource limitation may impose an upper limit to fruit
production in orchids, many species could sustain sig-
nificantly higher levels of pollination throughout their
lifetimes.
Many orchids face low fruit production as a result of
low pollination, but they are able to produce many
fruits if pollinated. If pollination is uncertain at the
individual level, then producing many fruits in a given
‘good’ year may be beneficial to the plant even if it
entails a cost, because the probability of being polli-
nated in the subsequent year may be very low, regard-
less of whether or not the plant sets fruit in the
current year (Zimmerman & Aide, 1989; Calvo & Hor-
vitz, 1990). For example, the lady’s slipper, Cypripe-
dium acaule, produces a single flower per year and
consistently exhibits strikingly low fruit set in closed
canopy forests where the orchids are usually found
(below 10%; Davis, 1986; Gill, 1989). Fruiting does
have a significant cost, but only after four consecutive
years, which may occur with a probability of 0.0001
(Primack & Hall, 1990). On the rare occasion that the
forest canopy opens up after a fire, pollinator activity
abruptly increases as does fruit set (Gill, 1996). This
sit-and-wait strategy probably pays off because the
long intervals of low activity allow plants to replenish

their resources before another fruiting event takes
place.
ECOLOGICAL EFFECTS ON REPRODUCTIVE
SUCCESS
Variation in the probability of a given flower setting
fruit is influenced by a number of factors. Phenology,
inflorescence, habitat, plant density, population size,
and temporal variation (Kindlmann & Balounová,
2001) may affect reproductive success as well as the
composition of the surrounding plant community.
PHENOLOGY
The degree of synchronization between flower anthe-
sis and pollinator availability may affect visitation
rates. In several temperate orchid species, individu-
als that flower early in the spring are pollinated at a
higher rate than those that flower later, as in
Calypso bulbosa (Ackerman, 1981), Orchis morio
(Nilsson, 1984), Orchis mascula (Nilsson, 1983b) and
Platanthera blephariglottis in Michigan (cf. Cole &
Firmage, 1984). The same phenomenon was observed
in the tropical epiphyte Tolumnia variegata (Sabat &
Ackerman, 1996). However, constant rates of pollina-
tion throughout the season were reported for the
same species of Platanthera in Maine (Smith &
Snow, 1976), and for the tropical epiphyte, Psychilis
krugii (Ackerman, 1989), which flowers all year. In
Dactylorhiza sambucina the highest rates of visita-
tion and pollinia removal occurred at the beginning
of the flowering season (Nilsson, 1980), but this
observation was not discussed in terms of fruiting
success.
Zimmerman et al. (1989) reported asynchronous
phenologies of Catasetum viridiflavum and its
euglossine bee pollinator, Eulaema cingulata (Fabri-
cius) in Panama. Peak flowering occurred up to six
months later than the peak of pollinator activity, with
fruit set of early flowering plants being about three
times higher than later flowering ones. However, early
initiated fruits were shed at a higher rate, even when
they appeared to be fully developed. This shedding of
fruits was presumably related to the long time fruits
have to be retained until seed dispersal, which occurs
during the next dry season. Zimmerman et al. (1989)
discussed the possibility of conflicting selective pres-
sures on the flowering time of C. viridiflavum. In sev-
eral cases, the phenology of orchids and their
pollinators has been recorded (Smith & Snow, 1976;
Nilsson, 1978b, 1983a; Ackerman, 1983; Inoue, 1985;
Nilsson et al., 1985), but the effects of individual vari-
ation in flowering phenology on fruit set was not fully
discussed.
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
EVOLUTIONARY PROCESSES IN ORCHIDS 27
INFLORESCENCE EFFECTS
Differential fruit set can occur, as a result of the posi-
tion of the flower on the inflorescence (Nilsson, 1980,
1983b, 1984; Cole & Firmage, 1984; Berry & Calvo,
1991; Vallius, 2000). In all reported cases, fruit set was
highest toward the bottom of the inflorescence and
decreased steadily toward the top. In Orchis morio,
O. mascula, and Dactylorhiza sambucina, which are
nectarless orchids with acropetal floral development,
the reduction in fruit set along the inflorescence was
dramatic. Fruit set of flowers at the top was between
50 and 90% lower than that of those at the bottom
(Nilsson, 1978a, 1980, 1983b, 1984; Vallius, 2000). A
similar pattern was observed in the nectar producing
Oreorchis patens in Japan (Sugiura, Okajima &
Maeta, 1997). These patterns may result from pollina-
tor foraging behaviour. Pollinators tend to move
upwards along inflorescences in order to forage for
nectar (Ackerman, 1975; Best & Bierzychudek, 1982);
they may depart after unsuccessfully probing a few
flowers, thus resulting in reduced pollination of flow-
ers located towards the top (Nilsson, 1980, 1983b).
It is possible that the nectar reward is insufficient to
encourage multiple flower visits or that the top flowers
are smaller in size, thus affecting reproductive success
(Sugiura et al., 1997). Alternatively, in reward species,
pollinators may lose interest as rewards in the popu-
lation diminish below a certain threshold as happens
with pollinators of other plant populations (Heinrich,
1975). In the case of deceptive species, the pool of
naive pollinators may become exhausted so that flow-
ers emerging in the latter half of the season would
receive few visitations (see ‘Phenology’ above). Fruit-
ing would therefore be more common on the lower and
older parts of the inflorescence.
A unique situation has been described for the high
altitude Andean orchid, Myrosmodes cochleare (Berry
& Calvo, 1991). As in other orchids, fruit set decreases
markedly toward the top of the inflorescence; however,
in this case flowers open in a basipetal sequence (i.e.
top flowers open first) and flower size increases
steadily from the top to the bottom of the inflores-
cence. Controlled pollination revealed that the ability
to set fruit decreases from the bottom to the top, and
appears to be associated with the change in flower
size. Because pollen viability is high along the inflo-
rescence, Berry & Calvo (1991) argued that
M. cochleare is functionally andromonoecious.
Position-related fruit set is common in other fami-
lies, but it has been generally explained in terms of
competition for resources among developing ovaries
(Lee, 1988). Basal, early initiated fruit may pre-empt
resources that could otherwise be allocated to other
fruit (Stephenson, 1981; Bawa & Webb, 1984). Thus, in
resource-limited species, position-related fruit set is a
28 R. L. TREMBLAY ET AL.
consequence of differential flowering or fruit abortion.
Orchids, however, are largely pollination-limited and
the pattern seems to be the result of differential pol-
lination along the inflorescence. Consequently, the
commonness and influence of resource allocation
needs to be studied and could explain position-
dependent fruit set, especially in terrestrial, many-
flowered species.
HABITAT AND COMMUNITY EFFECTS
Another potential source of variation in attractiveness
to pollinators (and thus in fruit set) among individuals
of a population is the microhabitat they occupy. In gen-
eral, plants that attract their pollinators by means of
visual cues may be less visited if they grow among
other plants or in shady spots (e.g. Orchis galilaea;
Bino, Dafni & Meeuse, 1982). Reduced fruit set has
been attributed to ‘closed habitat’ in Platanthera cili-
aris (Smith & Snow, 1976), and P. blephariglottis (Cole
& Firmage, 1984), where plants were somewhat hid-
den by the surrounding vegetation. In Cephalanthera
longifolia, fruit set of plants located in shady micro-
habitats was 40–50% lower than that of those in
sunny areas in two consecutive years (Dafni & Ivri,
1981b). Similarly, M. Morales & J. D. Ackerman
(unpubl. data) found that in a population of the epi-
phytic Tolumnia variegata, plants in the sun had 20%
fruit set whereas those in the shade had only 3%.
Inoue (1985) studied two moth-pollinated species of
Platanthera; in one there was a detectable reduction
in fruit set related to plant cover, but in the other no
relationship was detected.
Sometimes fruiting success of an individual may
simply depend on whether it is located where its pol-
linators are present. Ackerman et al. (1997) found that
population variation in fruit set of Tolumnia variegata
was associated with the abundance of their Centris
bee pollinators. Nilsson (1983c) noted that fruit set in
Cephalanthera rubra, which deceives its solitary male
bee pollinator, Chelostoma spp., was 30% in areas that
were patrolled by the bees, and only 7% in unpatrolled
areas (Nilsson, 1983c). Finally, deceptive orchids may
have higher fruit set if the population overlaps with a
rewarding species and they share one or more pollina-
tors. Cypripedium acaule benefits from bumblebee
activity on interspersed Vaccinium (Gill, 1996). Dafni
& Ivri (1981a) observed that fruit set of the nectarless
Orchis israelitica, which resembles flowers of the nec-
tariferous Bellevalia flexuosa Boiss. (Liliaceae), was
below 5% where Bellevalia was absent and higher
than 35% with it present. Similarly, fruit set in Orchis
caspia was 2–6 times higher where nectar producing
species of other families were present (Dafni, 1983).
However, fruit production of Epidendrum radicans
Pav. ex Lindl. was apparently unaffected by the pres-
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
ence of Asclepias curassavica L. and Lantana camara
L., two nectar producing species with similar floral
displays that share pollinators with it (Bierzychudek,
1981b), although other nectar resources were not
taken into account.
In the rewardless Orchis boryi, reproductive success
can depend on the flower colour of the plants in the
surrounding community (Gumbert & Kunze, 2001).
Bees that preferentially foraged on flowers similar in
colour to that of the orchid were more likely to visit the
orchid as well. Visits to O. boryi were also influenced
by the density of both the orchid and the surrounding
rewarding plants.
Reproductive success can vary dramatically between
adjacent sites where habitats differ. Sugiura et al.
(1997) observed that at an exposed seashore site in
Japan, the unpollinated flowers of Cypripedium mac-
ranthos lasted an average 5.1 days and fruit set was
8.3%. At a nearby sheltered area, longevity was twice
as long yet fruit set was only 1.2%. The authors suggest
that the lower fruit set was a direct consequence of
anthropogenic intervention (tourism, site two only:
1500 visitors/day) interfering with the pollinators.
DENSITY EFFECTS
Clumped distributions may result in higher pollina-
tion rates if pollinators respond to the larger floral dis-
play. However, not many studies have addressed this
possibility. In Calopogon tuberosus, clumps of interme-
diate size (2–8 plants in a 1-m radius) were found to
have higher fruit set than either solitary plants or
clumps of nine or more plants (Firmage & Cole, 1988).
More pollinators may be attracted to the larger
clumps, but because C. tuberosus has a deceptive pol-
lination system, they might be abandoned after a few
unrewarding visits, resulting in lower overall fruit set
(Firmage & Cole, 1988). No effect of clump size or
plant densities was found in Brassavola nodosa
(Schemske, 1980), Leporella fimbriata (Peakall,
1989a) or Malaxis massonii (Aragón & Ackerman,
2001). Plant densities also had no effect on reproduc-
tive success in Listera cordata except where plants
where highly dispersed (Meléndez-Ackerman & Ack-
erman, 2001). Alternatively, Schemske (1980) sug-
gested that pollinators of Brassavola nodosa may
focus on the inflorescence as the unit of attraction
instead of clusters of inflorescences. This is corrobo-
rated by some studies (Montalvo & Ackerman, 1987;
Aragón & Ackerman, 2001), but not by others
(Meléndez-Ackerman & Ackerman, 2001). In one case
Sabat & Ackerman (1996) found that frequency of vis-
itation to a population of Tolumnia variegata was
more closely related to the number of flowers on a host
tree than to the number of flowers on a host branch or
orchid inflorescence.

POPULATION SIZE EFFECTS
Population size can have an effect on reproductive suc-
cess. For orchids, the distribution of population sizes is
skewed, with most populations holding only a few
individuals and few populations are large (Whigham
& O’Neill, 1988; Tremblay, 1997c). Fritz & Nilsson
(1994) observed the effect of pollinaria removal and
fruit set in populations of differing size in three spe-
cies of deceptive orchids in Sweden (Orchis spitzelii,
N = 10; Orchis palustris Jacq., N = 8, and Anacamptis
pyramidalis, N = 10). The number of pollinaria remov-
als and fruit set increased with population size, but
the proportion of pollinaria removed per plant
decreased. The effect on fruit set was inconsistent
among the three species: a reduction was observed in
the butterfly-pollinated A. pyramidalis but no correla-
tion was observed in the other two species. In general,
larger populations had: (1) more pollinaria removed
and higher fruit set; (2) decreased pollinaria removals
per individual; (3) increased variance in male (but not
female) reproductive success; (4) an increased ratio of
pollinaria removal to fruit set; and (5) increase in vari-
ation of pollinator types.
In addition to Fritz & Nilsson’s (1994) study,
Donaldson et al. (2002) examined the effect of habitat
fragmentation on pollinator diversity and reproduc-
tive success and showed that these factors can influ-
ence reproductive success by demonstrating that
isolated small populations can fail to set fruit. Murren
(2002) investigated the effect of fragmentation on
reproductive success in Catasetum viridiflavum in ten
islands created by the Panama Canal and five nearby
mainland sites. Reproductive success, as measured as
fruit set, was significantly lower on islands as com-
pared to mainland populations in two of three years.
We have supplementary data on the number of pol-
linations or fruits set and variance in male and female
reproductive success. Ackerman (1981) found the
number of fruits set in the deceptive Calypso bulbosa
var. occidentalis was strongly related to population
size [linear regression, N = 7, r2 = 0.84, P = 0.004 (our
analysis)] as did Peakall (1989a) for the number of pol-
linations in populations of Leporella fimbriata [N = 16,
r2 = 0.46, P = 0.004 (our analysis)]. However, Peakall
(1990) did not find such a relationship for Drakaea
glyptodon [N = 9, P = 0.3 (our analysis)]. Fritz & Nils-
son (1994) showed that only variance in female repro-
ductive success was unrelated to population size
whereas Tremblay (unpubl. data) found that variance
in both male and female reproductive success in three
species of Caribbean Lepanthes was independent of
the size of populations (all P > 0.20).
Although we often find a positive relationship
between population size and the number of pollina-
tions or fruits set, per cent fruit set shows no such
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
EVOLUTIONARY PROCESSES IN ORCHIDS 29
relationship. Per cent fruit set was independent of
population size in the rewardless Calypso bulbosa var.
occidentalis [Ackerman (1981) linear regression,
r2 = 0.22, N = 7, P = 0.61 (our calculations)], although
there was a tendency for reduced per cent fruit set in
very small or very large populations. Alexandersson &
Ågren’s (2000) similar study of the same species noted
a negative relationship between pollen removal and
population size, although the pattern was only
observed in one of two years. Moreover, fruit set was
population size independent. Similarly, in populations
of the sexually deceptive Australian terrestrials, Dra-
kaea glyptodon (N = 9), Caladenia tentactulata (N = 5)
and Leporella fimbriata (N = 16), the percentage of
flowers pollinated was independent of population size
[linear regressions: all P ≥ 0.3 (our calculations)] but
there was a trend for the largest populations to have
lower rates of pollination (Peakall, 1989a, 1990;
Peakall & Beattie, 1996). Per cent fruit set of South
African populations of sexually deceptive Disa atrica-
pilla (N = 6, P = 0.39) and D. bivalvata (N = 4,
P = 0.28) were again unrelated to population size
(Steiner, Whitehead & Johnson, 1994). Furthermore,
pollinator behaviour may be affected by population
size variation which may itself be density dependent
(Gillman & Dodd, 2000). Thus, in general, reproduc-
tive success as measured by per cent fruit set or flow-
ers pollinated in populations of deceptive species in
Africa, Australia, North America and the Caribbean,
appears to be unrelated to most population sizes
encountered.
TEMPORAL VARIATION IN REPRODUCTIVE EFFORT
Fruit set in some populations of both tropical and tem-
perate orchids is consistent from year to year. Popula-
tions of Orchis mascula showed 4.8–7.1% fruit set over
a period of 6 years (Nilsson, 1983b), and V. C. Quesnel
(pers. comm.) reported that fruit set was consistently
low in a Trinidad population of Cyrtopodium parviflo-
rum Lindl. (1978, 10%, 1979, 1%, 1980, 3% and 1981,
8%). Fruit set was also consistent in Epidendrum cil-
iare and was even unaltered after a hurricane severely
altered the habitat (Ackerman & Moya, 1996).
Consistency may commonly exist, but not all popu-
lations of a species behave in a similar fashion. Tol-
umnia variegata fruit set differed ten-fold among
Puerto Rican populations, but was consistent among
years (Ackerman et al., 1997). Usually, Cypripedium
acaule also has a fairly constant fruit set (Gill, 1989;
Primack & Hall, 1990) and a low one too, but unlike
E. ciliare, a large increase in fruit production occurred
when the environment was radically modified. After
an outbreak of gypsy moth caused trees above the
orchids to defoliate, the increased light brought a dra-
matic increase in flower production in orchids and
30 R. L. TREMBLAY ET AL.
blueberries. The latter were an important nectar
source for bees, pollinators of both the blueberries and
the orchids. The bee populations increased, and orchid
fruit set leaped to 20% (Gill, 1996). Such environmen-
tal perturbations may be responsible for the variable
fruit set reported in other orchid populations such as
Epipactis palustris (5.01–19.80% in 3 years; Nilsson,
1983b) and Orchis morio (4.3–30.2% over a 5-year
period; Nilsson, 1984).
EVOLUTIONARY PROCESSES AND
CONSEQUENCES
Studies of patterns of evolutionary divergence in
orchids have flourished over the last decade, with
numerous papers addressing evolutionary relation-
ships among species, genera, tribes and subfamilies of
the Orchidaceae (Benzing, 1987; Chase & Palmer,
1997; Barkman, 2001; Barkman & Simpson, 2002;
Mant et al., 2002). These studies have generated much
discussion, spawning hypotheses of evolutionary pro-
cesses to explain the patterns of relationships. How-
ever, our understanding of the processes by which
orchids evolve is still in its infancy. In this section we
discuss how variation originates in orchids, describe
the evolutionary processes associated with these
plants and review the evidence for each.
SOURCES OF VARIATION
Evolution can only occur if there is variation among
individuals. The most commonly acknowledged cause
of variation is point mutation. Using phylogenetic
data, Barraclough & Savolainen (2001) estimated neu-
tral molecular evolution in flowering plants and dem-
onstrated that a positive relationship between species
number and rate of molecular change exists. What is
the cause of molecular rate change? Barraclough &
Savolainen (2001) suggested that three possible com-
ponents could influence and cause higher evolutionary
rates: short generation time, small population size and
high mutation rates. Mutation rates may vary among
different plant lineages for a number of reasons,
including mismatch repair efficiency and mutagen
exposure (Merrell, 1981; Gaut et al., 1996; Barra-
clough & Savolainen, 2001).
In addition to DNA mutation, variation is also
affected by meiotic recombination (Adelson, 2001). In
orchids, every fruit produces many seeds; the largest
contain over a million (Arditti & Ghani, 2000). Thus,
the opportunity for meiotic recombination is high,
where each new combination constitutes an experi-
ment in variation.
Another process influencing variation in orchids is
polyploidy and aneuploidy, both of which are not
uncommon. (Tanaka & Kamemoto, 1984). From a hor-
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
ticultural point of view, polyploidy often results in
more robust plants with flowers that are wider,
thicker, more erect, sturdy or compact. However,
Tanaka & Kamemoto (1984) note that whereas polyp-
loidy leads to distinct differences from parental plants,
aneuploidy rarely leads to differences from the paren-
tal plant except for deviations from the basic chromo-
some number.
Hybridization generates considerable variation and
is thought to be a major reason for plant diversifica-
tion (Stebbins, 1959; Campbell Waser & Meléndez-
Ackerman, 1997). In fact, coupled with polyploidy, it
may lead to sympatric speciation of allopolyploidic
populations (Hedrén, 1996). Natural hybrids occur
among orchids and new examples are reported every
year. However, convincing genetic evidence is rarely
presented for putative hybrids. Hybrid swarms and
hybrid origins of species appear to be commonplace in
Europe (Scacchi, De Angelis & Lanzara, 1990) but are
rarely reported elsewhere. The high frequency in
Europe may be overstated (Hedrén, 1996) whereas the
low occurrence elsewhere may be understated. In at
least one neotropical genus, Tolumnia, a number of
processes seem to be operating that generate variation
and these do include both hybridization and poly-
ploidy (Withner, 1976; Braem, 1988).
Whatever the origin of variation within and among
orchid populations, the consequences for orchid evolu-
tion depend on the same processes. Rather than
review and debate the relative importance of various
sources of variation, we focus on the conditions and
processes that shape that variation.
NATURAL SELECTION AND GENETIC DRIFT
Two processes can lead to evolution: natural selection
and genetic drift. Natural selection is a process that
requires (a) variation in character states among indi-
viduals (variation); (b) a relationship between trait
variants and the ability of the individuals to leave off-
spring (fitness differences); and (c) that the trait be
heritable between parents and offspring (inheritance).
Genetic drift requires (a) and (c), but (b) is absent
(Endler, 1986). Population sizes necessary for natural
selection and genetic drift to occur are likely to be dif-
ferent. Natural selection is more likely in populations
with large effective sizes, unless the selection coeffi-
cient is high, while genetic drift is more likely in pop-
ulations with small effective sizes. The presence of
natural selection does not exclude genetic drift and
vice versa. It is important to appreciate that in small
populations, small selection coefficients are effectively
neutral: the smaller the population, the larger the
selection coefficient must be to overcome genetic drift.
Consequently, for natural selection to be dominant
and observable in small populations, the selection

coefficients need to be high (Merrell, 1981; Tremblay,
1997a).
Are fruit set and pollinaria removals good
indicators of fitness?
Attempts to understand the reproductive ecology of
orchids and flowering plants in general have focused
on fruit and seed production. Arguments about polli-
nator vs. resource limitation, pollinator efficiency and
fitness have been made under the assumption that if
fruit set is higher, then fitness would be higher. Evo-
lutionary arguments about pollinator limitation and
resource limitation are applicable only to the extent to
which fruit production affects overall fitness and is
consequently subjected to selection. The only study to
date that addresses this question in orchids is Acker-
man, Sabat & Zimmerman (1996), who found that
higher fruit production in Tolumnia variegata
resulted in an increase in seedling establishment.
Thus, in one of 20 000 or so species of orchids, fruit set
is a good indicator of reproductive success and, in par-
ticular, female fitness.
Occasionally the number of pollinaria removals is
used as a measure of male fitness; again, however, we
have found only one study that has tested this notion.
Nilsson et al. (1992) used microtags on pollinaria of
Aerangis ellisii and found that there was a very strong
relationship between the number of pollinaria
removed and success as a pollen donor.
Fitness is a measure of the relative reproductive
success of individuals (fruit set or pollinaria removals)
as compared to the mean of the population. Conse-
quently, it is the ability of an individual to leave off-
spring as compared to others, making the number of
fruits produced or pollinia donated important. On the
other hand, per cent fruit set is a measure likely to be
more relevant for comparing growth among popula-
tions.
EVIDENCE OF NATURAL SELECTION
There is no doubt that selection is responsible for
many (probably most) of the floral adaptations of
orchids. However, there are few examples that show
how natural selection occurs in orchids and the
strength and direction of the selection. We describe
some of the evidence in this section.
Influence of inflorescence display size on
reproductive success
Inflorescence size (number of flowers per inflores-
cence) or display size (number of flowers open) varies
in most orchids. If reproductive success is associated
with size or display, then the number of flowers pro-
duced may be affected by selection. Plants with larger
inflorescences had a higher probability of setting fruit
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
EVOLUTIONARY PROCESSES IN ORCHIDS 31
in Brassavola nodosa (Schemske, 1980), Lepanthes
wendlandii (Calvo, 1990b), Calopogon tuberosus (Fir-
mage & Cole, 1988), Ionopsis utricularioides (Mon-
talvo & Ackerman, 1987) and Aspasia principissa
(Zimmerman & Aide, 1989). This was not the case with
display size in the self-incompatible Psychilis krugii
(Ackerman, 1989), Cypripedium fasciculatum (Lipow,
Bernhardt & Vance, 2002) or inflorescence size in Epi-
dendrum exasperatum (Calvo, 1990b). However, pro-
longed sequential production of flowers in Psychilis
monensis improves probability of both male and
female success (S. Aragón, unpubl. data).
The effect of inflorescence size on fruit production
can be time dependent. Rodríguez-Robles et al. (1992)
found a positive relationship in Comparettia falcata in
one year but not in another. Furthermore, response to
variation in inflorescence size may differ among polli-
nators. Inoue (1986a) showed in a study of the moth-
pollinated Platanthera metabifolia that species of the
family Sphingidae were clearly the most effective pol-
linators and responded positively to inflorescence size,
whereas those of the Noctuidae were poor pollinators
and did not favour the larger inflorescences. Maad
(2000) found display size to be highly significant and
directional in P. bifolia in three consecutive years.
Thus, flower production in most cases is positively
related to fitness so that one would expect selection for
increased flower production in the absence of any
other constraints.
Floral display, whether measured by inflorescence
or flower size, may differentially affect male and
female reproductive success. A role for male-male
competition in the evolution of display size assumes
that pollen is not limiting to reproduction (Stephenson
& Bertin, 1983; Willson & Burley, 1983). Therefore, in
pollen-limited taxa, one might expect that display size
would influence male and female reproductive success
equally. In those orchids that have been studied, this
appears to be true. In each of these studies, male
reproductive success was estimated from pollinaria
removals and female reproductive success was mea-
sured as fruit set (usually) or pollination, as deter-
mined by the presence of pollinia on the stigma. In the
tropical orchids Aspasia principissa (Zimmerman &
Aide, 1989), Psychilis krugii (Ackerman, 1989), and
Comparettia falcata (Rodríguez-Robles, Meléndez &
Ackerman, 1992) relative male and female reproduc-
tive success does not vary with inflorescence size and
is therefore parallel with respect to display size. How-
ever, these relationships can be site dependent.
Schemske (1980) reported a disproportionate increase
in both male and female reproductive success with
respect to inflorescence size in Brassavola nodosa, but
trends for male and female function were nonetheless
parallel. Murren & Ellison (1996) found that larger
displays in B. nodosa at another locality had dispro-
32 R. L. TREMBLAY ET AL.
portionately greater male than female reproductive
success. Similar results were observed for Psychilis
krugii (Ackerman, 1989).
The relationship between display size and male and
female reproductive success is equally complex for
temperate orchids. Piper & Waite (1988) concluded
that secondary floral characteristics were male spe-
cialized in Epipactis helleborine because pollinia
removals were greater than depositions. However, as
pointed out by Snow & Whigham (1989), trends in
estimates of male and female reproductive success
across the range of inflorescences were largely paral-
lel, indicating no differential effect of this aspect of
display. They also cited unpublished data from Tipu-
laria discolor supporting the same conclusion. Fir-
mage & Cole (1988) suggested that inflorescence size
of Platanthera blephariglottis influenced male and
female reproductive success in a parallel fashion over
small inflorescences (1–4 flowers) but that female
reproductive success levelled off with increasing inflo-
rescence size while male reproductive success contin-
ued to increase. However, samples of large
inflorescences were few and it is not clear that this
trend was real.
The relationship between inflorescence size and
male and female reproductive success in temperate
and tropical orchids appears to suggest parallel
response for most species with few exceptions. More-
over, we agree with Nilsson (1992) that clarification of
this relationship may be dependent on whether flow-
ers develop synchronously or sequentially. Further
research is required.
Some constraints on floral display
Inflorescence size may be influenced by multiple fac-
tors over ecological and evolutionary time. Total flower
production is dependent on plant size in many plant
species (Harper, 1977) including orchids (Montalvo &
Ackerman, 1987; Zimmerman & Aide, 1989; Calvo,
1990a). We have shown that larger inflorescences of
orchids generally have greater male and female repro-
ductive success. However, plant size distribution in
many populations is strongly hierarchical, with the
majority of the individuals being small (Weiner & Sol-
brig, 1984; Gregg, 1991b; Leeson, Haynes & Wells,
1991). Consequently, most plants may have relatively
small inflorescences (e.g. Schemske, 1980; Firmage &
Cole, 1988). Even if larger inflorescences are benefi-
cial, an increase in the mean inflorescence size of a
population may be limited by energetic and allometric
constraints.
Another factor that may impose limits on inflores-
cence size is the frequency of geitonogamous pollina-
tion (Ackerman, 1989), which may be greater in larger
inflorescences and result in higher rates of flower and
fruit abortion due to self-incompatibility or inbreeding
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
(e.g. Wyatt, 1982; Hessing, 1988; Klinkhamer & de
Jong, 1993). There are numerous mechanisms which
enhance the probability of cross-pollination in orchids
and manipulation of display size is one of them. Some
orchids have small floral displays yet produce many
flowers over long periods (e.g. Lepanthes and Psychilis
spp.). Displays at any given moment may be small, but
total flower production in these plants is positively
related to reproductive success (Lepanthes spp., R. L.
Tremblay, unpubl. data; Psychilis monensis, Aragón &
Ackerman, 2004).
Although reproductive success in orchids is strongly
pollen limited, resource availability plays a major role
in affecting display size. We could suggest that there
should be selection for more flowers, but the action of
that selection may be for efficient procurement and
storage of resources for reproductive events. There is
scope here, too, for further investigation.
Effect of flower size on reproductive success: The
number of flowers produced is one component of dis-
play and flower size is another. Larger flowers may be
a more powerful signal to pollinators than smaller
ones and may be affected by selection in a similar
manner to flower production. We found five studies
that compared flower size with reproductive success.
Murren & Ellison (1996) showed that larger flowers
had higher male reproductive success in Brassavola
nodosa. However, male and female reproductive suc-
cesses were independent of flower size in Tolumnia
variegata (Sabat & Ackerman, 1996), Myrmecophila
tibicinis (Malo, Leirana-Alcocer & Parra-Tabla, 2001)
and Cypripedium acaule (O’Connell & Johnston,
1998).
Due to the near ubiquity of pollen-limited orchid
reproduction, one would expect no differential effect of
flower size on male and female reproductive success.
However, more experimental studies should be con-
ducted, perhaps utilizing modification of display size
as in Malo et al. (2001) to show whether or not this is
true. If flower size indeed influences male and female
reproductive success equally, then selection to ensure
pollinator visits should be higher than in species
where male and female reproductive success is
unequal. As a result of pollinator limitation selection
would favour the showiest variants in a population of
orchids.
Spur characteristics and reproductive success: Spur
length, width of spur mouth and nectar reward should
be closely correlated with reproductive success. Dar-
win recognized that effective pollination is dependent
on the coupling of the morphology of the flower with
that of the vector. One of the most commonly given
examples of adaptation is the extraordinary spur
length of Angreacum sesquipedale Thou. and the
matching proboscis length of the hawkmoth pollinator

Xanthopan morgani (Walker) ssp. praedicta. Variation
in spur length and spur-mouth width has been impli-
cated in orchid evolution and reproductive isolation
(e.g. Nilsson, 1980, 1983a, b, 1985, 1988). While there
is little debate that the spur length of orchids is adap-
tive, recognition of such features is not indicative of
the processes involved.
Most of the evidence relating natural spur length
variation to reproductive success is circumstantial. Its
effects have been evaluated in few species. Luyt &
Johnson (2001) failed to detect selection for length in
Mystacidium venosum, while Inoue (1986b) found that
in natural populations of Platanthera mandarinorum
ssp. hachijoensis, variation in length was independent
of reproductive success. However, he tested the effect
of spur-shortening and a simulation of spur elongation
and found that it caused a reduction in male and
female reproductive success. Inoue interpreted the
results to suggest that extreme variants are selected
against and that stabilizing selection is likely.
Nilsson (1988) also examined the importance of spur
length variation on male and female reproductive suc-
cess. For Platanthera bifolia and P. chlorantha he
showed that both male and female fitness increase
with spur length and that it should be under direc-
tional selection.
The most comprehensive approach to studying
selection in orchids was performed by Maad (2000).
She observed that long spurs of Platanthera bifolia
had a female advantage but the selection differential
was only observed in one of three years. Selection can
be variable not only across seasons, but within a sea-
son as well. Maad (2000) observed that a combination
of the time of flowering and the length of the spur
affected reproductive success. Plants with short spurs
were favoured through female function early in the
season while those with long spurs were favoured
later in the season.
Floral colour and reproductive success: Most pollina-
tors are sensitive to colour. Consequently, colour vari-
ation among orchids in a population should be
perceived by pollinators and the most attractive colour
morph should be preferentially selected. Such varia-
tion exists and can be substantial, particularly in
deceptive orchids (e.g. Dactylorhiza sambucina - Nils-
son, 1980; Orchis caspia - Dafni, 1983; Thelymitra epi-
pactoides - Cropper & Calder, 1990). Sometimes the
variation is continuous (Psychilis monensis - Aragón
& Ackerman, 2004) while at other times there are dis-
tinct colour morphs.
Have we found selection for flower colour? Few stud-
ies have analysed this problem and only two include
behavioural data of pollinators. Smithson (2001) and
Gigord, MacNair & Smithson (2001) found in Dacty-
lorhiza sambucina that there is a reproductive advan-
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
EVOLUTIONARY PROCESSES IN ORCHIDS 33
tage to the rare corolla colour morph of the
nonrewarding orchid. Smithson (2001) corroborated
her field studies with behavioural experiments in the
laboratory. Negative frequency-dependent selection
seems to occur for flower colour in her system.
Koivisto, Vallius & Salonen (2002) examined the
behaviour of pollinators on two colour morphs of
D. maculata and noted that bumblebees preferred the
darker coloured morphs while other visitors had no
preferences. In contrast, R. L. Tremblay (unpubl. data)
examined colour variation in Lepanthes rupestris pet-
als and found no evidence of differential selection in
natural populations.
J. D. Ackerman & W. Carromero (unpubl. data) mea-
sured reproductive success in two colour morphs
(magenta and white) of Bletia patula in the Dominican
Republic and discovered that there was no significant
relationship between flower colour and male or female
reproductive success. Nevertheless, there was a ten-
dency for pollinaria removals to be higher in the white
morph (P = 0.056). Aragón & Ackerman (2004) exper-
imentally manipulated flower colour variation in pop-
ulations of Psychilis monensis and detected negative
frequency-dependent selection in two of three trials.
Thus, flower colour in orchids can be under selection
but not necessarily all the time.
Community structure of flower colour can also affect
orchid reproductive success. Pollinators of Orchis
boryi react to the diversity of flowers in the immediate
vicinity of the orchid; the similarity of these (model)
plants can have a positive effect on the orchid’s repro-
ductive success. Orchis boryi had a higher reproduc-
tive success if pollinators foraged on flowers of similar
colour (Gumbert & Kunze, 2001).
Floral rewards and reproductive success: Nectar, fra-
grances, oils, pseudopollen, resins and waxes (van der
Pijl & Dodson, 1966; Dondon et al., 2002) and even pol-
len (Vogel, 1981 reported in Pridgeon et al., 1999;
Gregg, 1991c) have been identified as pollinator
rewards in orchids. Variation in these rewards is
expected to directly influence pollinator behaviour and
consequently affect plant reproductive success either
through frequency of visits or quality of pollination.
Nevertheless, among all pollinator attractants, we
perhaps know the least about variation in rewards
and its consequences.
Nectar is the most common pollinator reward
offered by orchids and is the only one in which vari-
ability has been associated with plant reproductive
success. Variation in the quantity of nectar exists
within inflorescences of Platanthera blephariglottis
but it is not consistently associated with the probabil-
ity of pollination (Cole & Firmage, 1984). Interplant
variation in nectar production was extensively studied
in Comparettia falcata by Rodríguez-Robles et al.
34 R. L. TREMBLAY ET AL.
(1992). They found no relationship between natural
variation in nectar availability and pollinator visita-
tion. However, when Ackerman, Rodríguez-Robles &
Meléndez (1994) experimentally removed nectar,
reproductive success declined. On the other hand,
when nectar was added, there was no effect, not even
an increase in geitonogamous pollinations (Salguero-
Faría & Ackerman, 1999). These results strongly sug-
gest that stabilizing selection may favour the meagre
nectar reward offered by C. falcata.
Perhaps the most intriguing story concerning nectar
production in orchids is actually the lack of it: approx-
imately one third of all species produce no pollinator
reward (Ackerman, 1986a). Visitors are deceived by
various means; most commonly the deception is based
on a general resemblance to food plants without any
specific mimicry. Naive pollinators visit a few flowers
before learning to avoid them (Heinrich, 1975).
When orchids are so pollinator-limited, how could
such a pollination system have evolved (Gill, 1989)?
Under pollen-limitation there should be strong selec-
tion to increase pollinator visits. What better way
could there be than to provide a more enticing reward?
Any plant carrying a mutation that enhances its
reward would likely be more successful than one with-
out it. This may have happened in the genus Disa.
Johnson, Linder & Steiner (1998) showed that nectar
reward systems in the genus are derived and evolved
independently three times. These results are quite
intriguing because they contradict the hypothesis that
deception systems are more specialized than reward-
based systems and are derived from them (Ackerman,
1986a; Dafni, 1987).
If we do indeed find that deceptive systems evolved
from rewarding ones in some lineages, then what
would have led to the loss of a reward? Selection may
have favoured a loss if having a reward resulted in (1)
high levels of geitonogamy (selfing) or (2), resource
depletion through high costs of nectar or fruit produc-
tion. Johnson & Nilsson (1999), Smithson & Gigord
(2001) and Smithson (2002) tested the geitonogamy
hypothesis by adding nectar to rewardless orchids
(Orchis spp., Barlia and Anacamptis). They found
that pollinator visitation increased dramatically as
expected, but that there was little or no effect on the
number of geitonogamous pollinations or total fruit
set except in Anacamptis. In the first two taxa
studied, delayed caudicle bending likely prevented
self-pollinations. As for resource depletion, we have
nothing but suggestive data: nectar production may
be expensive (Southwick, 1984; Koopowitz & March-
ant, 1998; Luyt & Johnson, 2002), and as already
reviewed, there is ample evidence of a physiological
cost to reproduction.
Smithson & Gigord (2001) identified a third possible
mechanism that would lead to evolution of a deception
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
system. In their nectar-addition experiment, they
found that nectarless plants had ten times more pol-
linaria removals as reward plants. If a population had
a nectarless mutant, then it would have a much higher
probability of fathering seed than a nectar-producing
plant. Such mutants would rapidly spread through a
population even when the effective population size is
small. Smithson & Gigord (2001) argued that a male
function advantage is the more likely explanation for
the evolution of deception systems.
Selection may not be the only avenue for the evolu-
tion or maintenance of deception. Nectar production
may have been lost through genetic drift if population
or display sizes were insufficient to attract or main-
tain pollinator interest regardless of reward quality or
quantity. Coincidentally, when population sizes are
small, both drift and competition for pollinator atten-
tion are more likely to occur. Tremblay & Ackerman
(2001) showed that conditions for drift do occur in
orchids, and the widespread pollen-limitation in
orchids implies that competition for pollinators is
severe. We still need experimental data on reproduc-
tive success across an array of population sizes to cor-
roborate this hypothesis.
Thus, we have phylogenetic evidence that nectar
producing orchids may have evolved from deceptive
species (Johnson et al., 1998) which is also supported
by theoretical considerations that deception should be
an unstable system (Gill, 1989). On the other hand,
there are data which indicate that deceptive species
may have evolved from reward species via costs to
reproduction or reward production (Ackerman & Mon-
talvo, 1990), or through male function advantage
(Smithson & Gigord, 2001). Perhaps there is no single
or dominant process by which deceptive pollination
has evolved or is maintained. However, the wide-
spread nature of the pollination system in orchids
argues against this possibility.
Odour and reproductive success: As in other charac-
teristics associated with pollinator attraction, floral
fragrances are usually considered to be adaptive. The
characterization of floral fragrances took immense
leaps in the 1960s and early 1970s, particularly for
orchids pollinated by male euglossine bees (Dodson
et al., 1969; Williams & Dodson, 1972). Field and
experimental studies have linked floral fragrances in
both temperate and tropical genera to specific pollina-
tors or groups of pollinators (Dressler, 1968; Hills, Wil-
liams & Dodson, 1972; Borg-Karlson, Bergström &
Kullenberg, 1987; Bergström et al., 1992; Alcock,
2000; Plepys, Ibarra & Löfstedt, 2002). Because the
emphasis was on reproductive isolating mechanisms,
characterization tended to be typological. It is cer-
tainly undeniable that interspecific variation in fra-
grances exists and that this may be attributed to

differences in pollinators. Nonetheless, there is con-
siderable intraspecific variation in floral fragrance
production and composition as well.
The first population-level sampling of orchid floral
fragrances that we know of showed that there was
considerable variation among plants in fragrances of
euglossine bee-pollinated Cycnoches species (Gregg,
1983). Within- and among-population variation was
also described in the European Platanthera bifolia
and P. chlorantha (Tollsten & Bergström, 1989, 1993).
In contrast, Patt, Rhoades & Corkill (1988) found
much quantitative but little qualitative variation in
populations of North American P. stricta. In a detailed
study of fragrance variation, Moya & Ackerman (1993)
showed that fragrances of Epidendrum ciliare are pro-
gressively more variable among flowers of an inflores-
cence, among inflorescences of a plant, among plants
of a population, and finally among populations. In
another study, some plants of the widespread Anti-
llean epiphyte, Tolumnia variegata, were found to be
fragrant whereas others were not (Ackerman, Melén-
dez-Ackerman & Salguero-Faría, 1997).
What are the causes of intraspecific variations in
floral fragrances? Some variation may be attributable
to hybridization (Tollsten & Bergström, 1993),
whereas genetic drift may be important in other cases.
Drift may come about due to founder events, small
effective population size of isolated populations (Toll-
sten & Bergström, 1993) or loss of function, as has
apparently happened for other characteristics
(Steiner, 1998). More recently it has been suggested
that variation in fragrance production and composi-
tion may be driven by natural selection, particularly
in species that employ deceptive means of attracting
pollinators. In the pseudocopulated Ophrys sphegodes,
part of the fragrance spectrum is invariant whereas
other parts are not. The invariant part mimics the
female sex pheromones of the pollinators while the
non-biologically active part of the fragrance is quite
variable. It is this variable portion of the fragrance
that impairs avoidance learning on the part of the
duped male bees (Ayasse et al., 2000).
Negative frequency-dependent selection has been
proposed to maintain high levels of variation in char-
acteristics associated with pollinator attraction for
deception-pollinated species (Ackerman & Galarza-
Pérez, 1991). Although negative frequency-dependent
selection on flower colour has been shown, we have not
been able to demonstrate it for fragrance. An attempt
was made to test this hypothesis by manipulating the
frequencies of fragrant and odourless plants of Tolum-
nia variegata but without success. As it turns out, the
fragrant plants are highly variable in fragrance com-
position (A. Cuevas, unpubl. data), a factor which may
have affected the outcome of the experiment. We are
unaware of any other attempt to associate reproduc-
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
EVOLUTIONARY PROCESSES IN ORCHIDS 35
tive success with odour variation in natural popula-
tions of orchids.
Because pollinators associate rewards with fra-
grances quite readily (Kunze & Gumbert, 2001), we
expect that some deceptive species would have an
odour identical to rewarding species, be odourless or
have a highly variable fragrance (Moya & Ackerman,
1993; Kunze & Gumbert, 2001). With the partial
exception of the Ophrys model (Ayasse et al., 2000), we
are unaware of any deceptive species with a fragrance
that mimics that of a rewarding model (Nilsson, 1980;
Kunze & Gumbert, 2001). Thus far, deceptive species
tend to be either odourless or quite variable.
Current evidence for natural selection on floral
odours is mostly indirect, coming from studies which
show that different fragrances are associated with dif-
ferent pollinators. Others have demonstrated the
adaptiveness of particular compounds (Schiestl &
Ayasse, 2001). In cases where variation is minimal, we
expect that the pattern would have been the result of
stabilizing selection. On the other hand, drift, nega-
tive frequency-dependent selection or hybridization
may be responsible for high levels of fragrance varia-
tion in some species.
Inflorescence height and reproductive success: How
high flowers are from the ground or substrate could
affect reproductive success if pollinators have foraging
height preferences. Handel & Peakall (1993) showed
that thynnine wasps preferred flowers of the sexually
deceptive Chiloglottis reflexa Druce that were lower to
the ground (15 cm) compared to flowers higher up in
the vegetation (55 and 105 cm). Maad (2000) found sig-
nificant selection differentials for stalk length in Pla-
tanthera bifolia in all three years of her study. However,
the selection differentials were more consistent
through the female function. O’Connell & Johnston
(1998) found that stalk length (flower height) was sig-
nificantly correlated with male and female reproduc-
tive success in Cypripedium acaule. In the Caribbean
genus Psychilis, inflorescences have an elongate scape
up to 2 m in some species. Aragón & Ackerman (2004)
detected a significant positive effect of height above
ground and length of inflorescence on reproductive suc-
cess for P. monensis but similar data failed to show any
such relationship for the related P. krugii (J. D. Ack-
erman & R. L. Tremblay, unpubl. data).
Multivariate selection
Selection can affect multiple characters simulta-
neously and can occur either as a result of characters
that are linked genetically or because of allometric
correlations. We found two studies that attempted to
measure multivariate selection in orchids and both
looked at phenological, vegetative and floral charac-
teristics. O’Connell & Johnston (1998) found negative
36 R. L. TREMBLAY ET AL.
correlational selection on plant height with flowering
time in one of two populations of Cypripedium acaule
whereas Maad (2000) found no evidence of correla-
tional selection in Platanthera bifolia. In the absence
of experimental manipulation of trait distributions,
future studies should consider using regression anal-
ysis as a powerful tool (Lande & Arnold, 1983) for
obtaining estimates of the strength of selection
acting both directly (univariate) and indirectly
(multivariate).
EVIDENCE OF GENETIC DRIFT
SHIFTING BALANCE THEORY OF EVOLUTION
While natural selection is the change in trait distribu-
tion across generations caused by selection of advan-
tageous characteristics, genetic drift is the change
resulting from random events. There are presently
two main hypotheses for evolution through genetic
drift: Wright’s shifting balance theory (Wright, 1968,
1969, 1977, 1978) and genetic drift including only the
two first steps of Wright’s theory (Coyne et al., 1997).
Wright’s shifting balance theory describes a mecha-
nism that involves genetic drift, followed by mass nat-
ural selection and migration among subpopulations. It
imagines an adaptive landscape with peaks and val-
leys of relative fitness. For example, peaks in fitness
may correspond to characteristics of flowers that pro-
mote the efficient deposition and receipt of pollen from
certain sized pollinators. Valleys are combinations of
floral traits that do not match available pollinators.
How then do members of a species pass from one
peak to another, thereby achieving reproductive isola-
tion and resulting in speciation? Wright (1978) pro-
posed three stages to his theory. First, relatively
isolated subpopulations or demes become genetically
differentiated through genetic drift, the stochastic
change in gene frequencies caused by small effective
population size (Ne). Put simply, Ne is the number of
individuals in a population actually leaving offspring
in the next generation. However, its meaning and
measurement are much more complicated than this
(Wright, 1939; Kimura & Crow, 1963; Waples, 1989;
1991; Wade, 1991; Barrowclough & Rockwell, 1993;
Nunney, 1993, 2000; Orive, 1993). Demes ‘drift’ ran-
domly into new genetic combinations and this process
is more likely with decreasing N e. When founding
events involve very few individuals colonizing a new
locale, extreme changes can occur, particularly if this
results in the alteration of genetic variance-covariance
matrices controlling the relationships between indi-
vidual genes (Carson, 1990). In this way, a deme may
pass or ‘skip’ fitness valleys, placing them on an oppo-
site slope of the adaptive landscape.
The second stage is mass selection, perhaps the
most straightforward aspect of the theory (Crow,
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
Engels & Denniston, 1990; Coyne et al., 1997). Once a
new variant has become established, one that achieves
some modicum of pollination efficiency from a new pol-
linator, selection will act to improve this efficiency.
Additive genetic variance allowing selection to occur is
assumed to be maintained by polymorphic mutation
and recombination (Kimura, 1965; Lande, 1975) and
may be augmented by the scrambling of genetic vari-
ance-covariance matrices during a founding event
(Carson, 1990; Roughgarden, 1996). Owing to overall
pollinator limitation, less fit individuals may repro-
duce infrequently if at all, speeding the fixation of pol-
lination efficient genotypes. As a concrete example,
Inoue (1986b) noted that the effect of spur length on
estimates of male and female reproductive success in
Platanthera mandarinorum was greater at lower pol-
lination frequencies than at higher ones. Thus, selec-
tion proceeds most rapidly under pollinator limitation,
again possibly distinguishing orchids from many other
plants if variance in reproductive success is higher or
more frequent in orchids.
The establishment of new pollination strategies of
closely related species with different pollinators may
be speeded up if only a few fruits are needed to set a
founder population. The fact that species of Catase-
tum can be distinguished by a few fragrance compo-
nents (Hills et al., 1972) may be the result of the quick
establishment of new genotypes. Thus, low fruit pro-
duction in orchids, coupled with the high number of
seeds produced per pod, may have provided a mecha-
nism for speciation and diversification in pollination
systems.
The third stage involves the movement of these
‘more’ fit genes back to the original populations. In
time, the different populations take on the new com-
bination of phenotypes from the original founding pop-
ulation (Wright, 1969; 1977; Lande, 1979; Coyne et al.,
1997). However, this process has never been docu-
mented in natural populations (Coyne et al., 1997).
Furthermore, it would not result in cladogenesis and
consequently cannot fully explain the great diversity
of orchids.
Commonality of small effective population size (N e )
We have argued and shown elsewhere (Zimmerman &
Aide, 1989; Calvo, 1990b; Ackerman & Galarza-Pérez,
1991; Tremblay, 1996, 1997a; Tremblay & Ackerman,
2001) that reproductive patterns in orchids may pro-
mote small Ne. Within a season, few flowering individ-
uals produce fruit and contribute to the seed pool,
particularly in populations of tropical orchids
(Table 3). Fruiting failure in a reproductive season is
high (Fig. 4). This may hold true even over longer sam-
pling periods because fruit production can suppress
flowering in subsequent years. Estimates of Ne for the
three species of the tropical epiphytic Lepanthes
 EVOLUTIONARY PROCESSES IN ORCHIDS 37

8 entiation through natural selection or genetic drift

(Merrell, 1981; Roughgarden, 1996). If the product of
Nm (the effective number of migrants per generation;
6 1
Fst = , N = effective population size, m =
4 Nm + 1
migration rate) lies between one and two, there will be
Frequency

4 considerable variation in gene frequencies among pop-

ulations (Merrell, 1981). The determining factor relat-
ing gene flow and natural selection is the effective
population size; the smaller it is, the more gene flow is
2
required to prevent local differentiation (Roughgar-
den, 1996). Thus, many small groups of individuals
with high gene flow would evolve as a unit if the sum
0
of the populations has a large N e. Consequently, nat-
0.

10

20

30

40

50

60

70

80

90
0–

.0

.0

.0

.0

.0

.0

.0

.0

.0
ural selection will likely dominate as the main evolu-
10

–1
.0

20

30

40

50

60

70

80

90

00
tionary process. Therefore, we need to know the
.0

.0

.0

.0

.0

.0

.0

.0

.0
Per cent fruiting failure
Figure 4. Distribution of fruiting failure in natural popu-
lations of orchids. Frequency of plants (N = 36) that fail to
set fruits. Data from Table 3.
revealed it to be below 20% for all species and popu-
lations; most were below 10% of the standing adult
population (Tremblay & Ackerman, 2001). Only 18%
of flowering individuals in a population of Aspasia
principissa produced fruit during the three years of a
survey by Zimmerman & Aide (1989). Furthermore,
the variances in female and male reproductive success
and in total lifespan are high and can further reduce
Ne. If the variance in recruitment is as skewed as the
previous variables, then Ne may be lowered even more.
While numerous factors contribute to N e, we believe
that these life history characteristics separate orchids
from many other plant groups and are all directly
linked to the extreme degree to which reproduction is
pollinator-limited. It will be necessary to investigate
in detail the commonality of small N e in orchids before
we can critically evaluate the frequency of this process
in orchid evolution.
Gene flow in orchids
Gene flow is an essential component of evolutionary
processes and critical to our argument that genetic
drift is an important component of diversification in
orchids. The amount of gene flow among local popula-
tions will determine whether or not individual popu-
lations (demes) can evolve independently and be
genetically distinct and plausibly lead to cladogenesis.
Theoretically, a gene flow rate greater than two indi-
viduals per generation is sufficient to prevent local
adaptation/evolution. Consequently, local selection
and drift will be overcome by migration (Merrell,
1981). Meanwhile, a flow rate less than one individual
per generation will likely result in population differ-
amount of gene flow among populations or groups of
individuals in order to identify the evolutionary pro-
cesses involved.
Gene flow and dispersal can be measured through
direct and indirect methods (Bohomak, 1999). Direct
observation of gene flow can be viewed by the use of
mark and recapture studies (for mobile organisms or
stained pollen) or tracking marker alleles (paternity
analysis) over a short number of generations. Indirect
methods involve obtaining allele frequencies and pro-
vide an estimate of the average long-term effect of dif-
ferentiation by genetic drift. The alleles are assumed
to be neutral so that differentiation based on these
markers would be a consequence of genetic drift
rather than natural selection. Bohomak (1999) con-
cluded that simple population genetic statistics
are robust for inferring gene flow among groups of
individuals.
Few orchid studies have attempted to directly
observe gene flow and thus far only staining or
microtagging of pollen have been used (Peakall,
1989b; Nilsson et al., 1992; Folsom, 1994; Tremblay,
1994; Salguero-Faría & Ackerman, 1999). All such
studies examined gene flow only within populations.
Current estimates of gene flow have been made
using indirect methods. They indicate that there is
much variation among species (Table 10). Data are
available for 70 species and subspecies of orchids. No
genetic differentiation among populations was
observed in two of the species. Twenty-nine species
with high levels of estimated gene flow (Nm > 2), 20 of
the species have intermediate to low levels of gene
flow (1 < Nm < 2) and 22 species have low levels
(Nm < 1; Fig. 5). There is much variation within a
genus, but there appears to be some consistency. For
example, Orchis species typically have high estimates
of gene flow among populations (Scacchi et al., 1990;
Corrias et al., 1991; Rossi et al., 1992) whereas Lepan-
thes spp. have much lower ones (Tremblay & Acker-
man, 2001). The present indirect approach to

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
38 R. L. TREMBLAY ET AL.

estimating Nm is actually the sum and interaction of
the historical and present gene flow (Slatkin & Bar-
ton, 1989).
Variation in gene flow estimates can be extensive,
even within the same species. In Cephalanthera
rubra, Scacchi et al. (1991) found high genetic popu-
lation substructure in Italy (FST = 0.245), whereas
Brzosko & Wroblewska (2003) found very little struc-
ture in South Korea (FST = 0.017). Similar large varia-
tions have been observed in Epipactis helleborine in
Europe and North America (Hollingsworth & Dickson,
1997; Scacchi, Lanzara & De Angelis, 1987; Squirrell
et al., 2001) and Cypripedium calceolus in North
America and Poland (Case, 1993, 1994; Brzosko,
Wróblewska & Ratliewicz, 2002; Brzosko, Ratkiewicz
& Wroblewska, 2003)
Are there phylogenetic associations with gene flow?
The Orchis and Lepanthes data are suggestive but
much more extensive sampling is needed for both tem-
perate and tropical species. Curiously, Lepanthes and
Orchis have very different population genetic param-
22
20
18
16
14
Frequency
eters yet both are species-rich genera and are likely in
a state of evolutionary flux.
EVOLUTIONARY DIVERSIFICATION MODELS
The commonness of pollinator limitation in orchids is
evident from the literature and we suggest it to be of
primordial importance in explaining evolutionary pro-
cesses in the family. The low levels of fruit set and the
skewed reproductive success and lifespan among indi-
viduals will result in small N e when gene flow among
populations is limited.
The presence of short and long-term resource con-
straints is not likely to have effected the importance of
pollinator limitation as a component of evolution in
the Orchidaceae because resource constraints have
mostly been observed between reproductive bouts and
not through fruit abortion. For example, if fruit abor-
tion occurred because of resource constraints, this
would result in a reduction in the variance in repro-
ductive success among individuals and an increase in
N e.
Interestingly, the process of natural selection in
orchids has rarely been detected in the field even
though it might seem obvious that it must have been
involved in the evolutionary process. Moreover, as
mentioned earlier, we are still ignorant of the strength
of natural selection, its frequency and directions. The
evidence we use to explain the great morphological

12
diversity that we assign to natural selection is either
10
8
pattern-based or circumstantial. We believe that the
6 great diversity of vegetative and floral characters
4 exhibited by the Orchidaceae is best explained by a
2 process that involves more than just natural selection.
0 In this section we will discuss our view of the process
0 2 4 6 8 10 12 14 16
Nm(W)
of evolution in orchids after differentiating natural
selection and Wright’s shifting balance theory and
Figure 5. Frequency distribution of gene flow estimates explain why these processes alone cannot account for
(Nm(W)) among populations of orchids. Data from Table 10. evolution in this family.

Table 10. Estimates of gene flow in orchids. Nm(S) = gene flow estimates (number of migrants per generation) based on
Slatkin’s private allele model; Nm(W) = gene flow estimates based on Wright’s statistics; GST coefficient of genic differen-
tiation among populations

Species References Nm(S) Nm(W) GST

Caladenia tentaculata Tate Peakall & Beattie, 1996 7.101 0.0346

Calypso bulbosa (L.) Oakes Alexandersson & Ågren, 2000 3.20 0.072
Cephalanthera damasonium (Mill.) Scacchi, De Angelis & Corbo, 1991 --5 --5
Druce
Cephalanthera longibracteata Blume Chung, Nason & Chung, 2004 0.762 0.247
Cephalanthera longifolia (L.) Fritsch Scacchi, De Angelis & Corbo, 1991 2.151 0.104
Cephalanthera rubra (L.) Rich. Scacchi, De Angelis & Corbo, 1991 0.761 0.247
Brzosko & Wroblewska, 2003 14.21 0.017

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
 EVOLUTIONARY PROCESSES IN ORCHIDS 39

Table 10. Continued

Species References Nm(S) Nm(W) GST

Cymbidium goeringii Rchb. f. Chung & Chung, 1999 8.48 2.30 0.098
Cypripedium acaule Ait. Case, 1994, 2002 1.271 0.164
Cypripedium arietinum R. Br. Case, 2002 --5 --5
Cypripedium calceolus L. Case, 1993, 1994 1.631 0.196
Brzosko, Ratkiewicz & 17.6 0.014
Wroblewska, 2002
Cypripedium candidum Muhl. Case, 1994, 2002 3.371 0.069
ex Willd.
Cypripedium fasciculatum Aagaard, Harrod & Shea 1999 6.00 0.04
Kellogg ex S. Watson
Cypripedium kentuckiense Case et al., 1998; Case, 2002 1.121 0.182
C. F. Reed
Cypripedium parviflorum Salisb. var. Case et al., 1998; Case, 2002 1.281 0.163
pubescens (Willd.) O. W. Knight
Southern populations Wallace & Case, 2000 0.94 0.209
Northern populations 1.57 0.137
var. makasin (Farw.) Sheviak var. 1.00 0.199
parviflorum species level 1.43 0.149
0.83 0.232
Cypripedium reginae Walter Case, 1994, 2002 0.471 0.349
Dactylorhiza romana (Sebastiani) Soó Bullini et al., 2001 3.321 0.07
Dactylorhiza sambucina (L.) Soó Bullini et al., 2001 1.311 0.16
Epidendrum conopseum R. Br. Bush, Kutz & Anderton, 1999 1.433 0.149
Epipactis helleborine (L.) Crantz Scacchi, Lanzara & De Angelis, 7.31 0.033
1987; Squirrell et al., 2001
European populations 1.001 0.200
0.241,4 0.5064
North American 2.531 0.0904
Hollingsworth & Dickson, 1997 0.791 0.240
Epipactis youngiana Richards & Porter Harris & Abbott, 1997 2.431 0.093
Eulophia sinensis Miq. Sun & Wong, 2001 -- 0.0
0.1331,3 0.6533
Gooyera procera Ker-Gawl. Wong & Sun, 1999 0.2211 0.523
0.3971,3 0.3863
Gymnadenia conopsea (L.) R. Br. Scacchi & De Angelis, 1989 0.2801 0.471
Gustafsson, 2000 3.917 0.06
Gymnadenia conopsea (L.) R. Br. Soliva & Widmer, 1999 2.96 0.078
conopsea
Gymnadenia conopsea (L.) R. Br. Soliva & Widmer, 1999 0.39 0.391
subsp densiflora (Wahl) E.G.
Camus & A. Camus
Gymnadenia odoratissima (L.) Rich Gustafsson & Sjögren-Gulve, 2002 1.066 0.19
Lepanthes caritensis Tremblay & W. Carromero, R. L. Tremblay & J. D. 0.23 1.30 0.167
Ackerman Ackerman, unpubl. data
Lepanthes eltoroensis Stimson Tremblay & Ackerman, 2001 1.54 0.89 0.220
Lepanthes rubripetala Stimson Tremblay & Ackerman, 2001 0.48 0.62 0.270
Lepanthes rupestris Stimson Tremblay & Ackerman, 2001 1.15 1.84 0.170
Lepanthes sanguinea Hook. W. Carromero, R. L. Tremblay & J. D. 1.74 1.45 0.144
Ackerman, unpubl. data
Lepanthes woodburyana Stimson W. Carromero, R. L. Tremblay & 0.61 7.5 0.032
J. D. Ackerman, unpubl. data
Leporella fimbriata Peakall & James, 1989 5.431 0.044

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
40 R. L. TREMBLAY ET AL.

Table 10. Continued

Species References Nm(S) Nm(W) GST

Nigritella rhellicani Teppner & Hedrén, Klein & Teppner, 2000 1.381 0.153
Klein
Orchis laxiflora Lam. Scacchi, De Angelis & Lanzara, 1990 2.851 0.08
Arduino et al., 1996 1.971 0.116
Orchis longicornu Poir. Corrias et al., 1991 4.20 12.252 0.02
Orchis mascula (L.) L. Scacchi, De Angelis & Lanzara, 1990 2.761 0.083
Orchis morio L. Scacchi, De Angelis & Lanzara, 1990 3.661 0.064
Rossi et al., 1992 4.751 0.05
Orchis palustris Jacq. Arduino et al., 1996 0.311 0.448
Orchis papilionacea L. Scacchi, De Angelis & Lanzara, 1990 6.331 0.038
Orchis pauciflora Ten. Scacchi, De Angelis & Lanzara, 1990 6.001 0.040
Orchis provincialis Balb. Scacchi, De Angelis & Lanzara, 1990 10.621 0.023
Orchis purpurea Huds. Scacchi, De Angelis & Lanzara, 1990 5.701 0.042
Orchis tridentata Scop. Scacchi, De Angelis & Lanzara, 1990 6.161 0.039
Paphiopedilum micranthum T. Tang Li, Luo & Ge, 2002 0.061 0.7977
& F. T. Wang
Platanthera leucopaea (Nutt.) Lindl. Wallace, 2002 0.081 0.754
0.711 0.263
Pleurothallis adamantinensis Brade Borba, Semir & Sheppard, 2001 4.851 0.049
Pleurothallis fabiobarrosii Borba Borba et al., 2001 2.841 0.081
& Semir
Pleurothallis johannensis Barb. Rodr. Borba et al., 2001 5.191 0.046
Pleurothallis ochreata Lindl. Borba et al., 2001 4.061 0.058
Pleurothallis teres Lindl. Borba et al., 2001 0.971 0.205
Pterostylis aff. alata M. A. Clem. Sharma et al., 2001 0.811 0.235
Sharma, Jones & French, 2003 4.381 0.054
Pterosylis angusta A. S. George Sharma et al., 2001 1.301 0.161
Pterosylis aspera D. L. Jones & M. A. Sharma et al., 2001 1.011 0.198
Clem.
Pterostylis gibbosa R. Br. Sharma, Clements & Jones, 2000 1.42 0.15
Pterostylis hamiltonii Nicholls Sharma et al., 2001 0.861 0.225
Pterosylis rogersii E. Coleman Sharma et al., 2001 1.101 0.186
Pterostylis scabra Lindl. Sharma et al., 2001 0.831 0.232
Spiranthes diluvialis Sheviak Arft & Ranker, 1998 5.44 0.044
Spiranthes hongkongensis S. H. Hu Sun, 1996 --5 --5
& Barretto
Spiranthes sinensis (Pers.) Ames Sun, 1996 0.53 1.19 0.174
Tipularia discolor (Pursh) Nutt. Smith, Hunter & Hunter, 2002 0.357 0.415
Tolumnia variegata (Sw.) Braem Ackerman & Ward, 1999 8.00 2.50 0.09
Vanilla barbellata Rchb. f. Nielsen & Siegismund, 2000 1.78 0.123
Vanilla claviculata (W. Wright) Sw. Nielsen & Siegismund, 2000 1.33 0.158
Zeuxine gracilis Blume Sun & Wong, 2001 0.5001 0.333
0.2141 0.5393
Zeuxine strateumatica Schltr. Sun & Wong, 2001 0.0213 0.9243

1
Nm calculated by the present authors from GST or FST using formula on p. 320 of Hartl & Clark (1989).
2
Recalculated using previous formula, original Nm value 3.70.
3
Calculated from RAPD markers.
4
Calculated from cpDNA.
5
No genetic differentiation found among populations.
6
Calculated according to Weir and Cockerham’s statistics.
7
Estimated using RAPD and AMOVA.

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54

DARWINIAN AND WALLACIAN GRADUALISM
Both Wallace (1858) and Darwin (1859) promoted the
view that change in character states among genera-
tions is gradual and driven by natural selection.
Armed with an understanding of gene expression, pop-
ulation genetics, and techniques to measure selection,
gradualism has come under fire from many quarters,
including orchid biologists.
Gradual change may occur when effective popula-
tions are large or gene flow is substantial among
smaller populations. Under these circumstances, most
genetic variation would occur within populations
rather than among them. We have some evidence,
mostly indirect, that such conditions do occur in
orchids. Pollen and seed dispersal is the usual means
for gene flow. Pollinators can move pollinaria long dis-
tances, presumably from population to population
(Janzen, 1971; Williams & Dodson, 1972; Ackerman &
Montalvo, 1985). The dust-like seeds are ideally suited
for wind dispersal (Carey, 1998; Murren & Ellison,
1998) and may be carried over great distances (Arditti
& Ghani, 2000). Nevertheless, fine-scale genetic struc-
turing (Chung, Nason & Chung, 2004), mathematical
models and wind tunnel experiments suggest limited
gene flow (Murren & Ellison, 1998). The rapid range
expansion of Oeceoclades maculata (Stern, 1988)
throughout the neotropics clearly demonstrates the
potential orchids have for dispersal. In addition, some
population genetic data are consistent with the high
gene flow hypothesis. For outbreeding species, varia-
tion among populations can be quite low, especially for
temperate terrestrial orchids (Table 10). If evolution-
ary change occurs in such populations, then it would
likely be a gradual process.
Nevertheless, we have also presented substantial
evidence that evolutionary change for many orchids is
not likely to be gradual. Effective population sizes may
be small because of a combination of factors: few
adults per population, pollinator limitation, skewed
reproductive success among individuals, and perhaps
skewed progeny recruitment. In such populations,
gene flow is restricted, and a high proportion of the
genetic variation occurs among populations. Gradual
evolutionary change is not consistent with those con-
ditions, so we advocate an alternative process that we
outline below.
THE DRIFT-SELECTION MODEL OF ORCHID EVOLUTION
A simpler scenario than the shifting balance theory is
the founding event–genetic drift scenario without the
third stage of Wright’s process. In this alternative
approach to evolution, genetic drift would be a
common occurrence in local populations followed by
natural selection, ultimately causing sufficient differ-
© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84, 1–54
EVOLUTIONARY PROCESSES IN ORCHIDS 41
entiation and the beginning of cladogenesis. Subse-
quently, this new taxon would colonize new sites.
Assumptions of this model include commonality of
small populations (low effective population sizes), low
or infrequent gene flow among populations, heritable
variation, and high or consistent selection coefficients
among generations in local populations.
Our present vision of orchid evolution involves both
genetic drift and natural selection working simulta-
neously. For most orchids, population sizes are small
and are effectively further reduced by skewness in
both reproductive success and lifespan of individuals.
Such conditions would likely result in frequent char-
acter changes among generations. Most of these
changes would be neutral or negatively selected with
the odd character having some positive selection
(directional, disruptive, stabilizing selection or some
non-parametric distribution). If true, then we should
observe large heritable character variation among
individuals, populations and generations.
The original evolutionary model for drift-selection is
Dodson’s (1962) ‘leap-frog’ model of orchid speciation
in Stanhopea. This genus is pollinated by relatively
small euglossine bees of the genus Euglossa and rela-
tively large ones of the genera Eulaema and Eufriesea
(Kimsey, 1982). Dodson proposed that speciation had
occurred repeatedly by the switching of species from
small to large pollinators and vice versa. This could
possibly occur through random changes in floral fra-
grances caused by genetic drift, followed by strong
selection on morphological characteristics to accom-
modate a different pollinator. This model was recently
given support by a study which demonstrated that the
molecular phylogeny of Stanhopea is not congruent
with phylogenies based on floral fragrances (Williams
& Whitten, 1999). A similar model of evolutionary
diversification may apply to the Australian genus
Chiloglottis and its Thynninae wasp pollinators. Mant
et al. (2002) showed that the pattern of pollinator
change is conservative in the sexually deceptive orchid
and that the phylogenies based on the DNA sequences
of the orchids and pollinators are highly congruent
(but not perfectly so). They emphasized that the
observed pattern is not evidence of coevolution
because relative branch length differences between
plant and insect groups were not similar. They also
suggested that pollinator shifts are dependent on the
availability of pollinators within the geographical
area, the phenology of flowering and wasp emergence,
and the changes in floral odour chemistry that mimic
female wasp pheromones. Thus, random changes in
floral fragrances could attract a different species of
wasp and lead to cladogenesis.
It is unlikely that we will observe in the field a
series of evolutionary events reflecting the totality of
the drift-selection model. However, it is possible to
42 R. L. TREMBLAY ET AL.
assemble sufficient field observations and experi-
ments to reconstruct the process. The model may be
supported by demonstrating that: (1) gene flow among
populations is restricted in time or space; (2) effective
population sizes are usually (or occasionally) small,
and (3) selection is episodically intense and different
among populations.
CONCLUSIONS
In this review, our central question has been: What
aspect(s) of orchid reproduction explain(s) the intri-
cate pollination mechanisms of this family? We have
argued that pollen-limitation of both male and female
reproductive success, in combination with the unique
structure of the orchid flower (the column), largely
explains the extravagant pollination mechanisms.
Low reproductive success, leading to a small propor-
tion of reproducing individuals and low N e, makes
genetic drift a more likely source of evolutionary
change in orchids that, combined with episodic selec-
tion, explains much of the diversification of the family.
Paradoxically, it is the low reproductive success of
orchids, not the opposite, that may account for both
their unique pollination mechanisms and extreme
diversity. The Orchidaceae is a remarkable group of
plants and the fundamental processes by which the
family has diversified are far more fascinating than
Darwin ever suspected.
ACKNOWLEDGEMENTS
This review was begun in the early 1990s by JKZ and
RNC, then abandoned as careers changed. They thank
RLT and JDA for reviving the project and achieving
levels of understanding never contemplated. JKZ
wishes to thank the support of the Smithsonian Envi-
ronment Research Center and Dennis Whigham dur-
ing the incipient phase of development. We greatly
appreciate critical comments from W. M. Whitten, N.
W. Williams and two anonymous reviewers. We also
thank all our colleagues who have contributed unpub-
lished data in the many areas of orchid biology.
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