Plant Biology ISSN 1435-8603

RESEARCH PAPER

Pollination and seed dispersal of Melocactus ernestii Vaupel

subsp. ernestii (Cactaceae) by lizards: an example of double
mutualism
V. G. N. Gomes1, Z. G. M. Quirino2 & I. C. Machado1

s–Graduacß~
1 Programa de Po ao em Biologia Vegetal, Departamento de Bot^anica, Centro de Ci^encias Biolo
gicas, Universidade Federal de Pernambuco, Recife,
Pernambuco, Brazil
2 Departamento de Engenharia e Meio Ambiente, Universidade Federal da Para
ıba, Rio Tinto, Para
ıba, Brazil

Keywords ABSTRACT
Caatinga; dry forest; inselberg; phenology;
saurochory; saurophily. Recent studies show that the mutualistic role of lizards as pollinators and seed dis-
persers has been underestimated, with several ecological factors promoting such
Correspondence plant–animal interactions, especially on oceanic islands. Our aim is to provide a quan-
I. C. Machado, Departamento de Bot^ anica, titative assessment of pollination and seed dispersal mutualisms with lizards in conti-
Centro de Ci^
gicas, Universidade
encias Biolo nental xeric habitats. We carried out focal observations of natural populations of
Federal de Pernambuco, CEP 50372–970, Melocactus ernestii (Cactaceae) in the Caatinga, a Brazilian semiarid ecosystem, in
Recife, Pernambuco, Brazil. order to record the frequency of visits, kind of resource searched and behaviour of vis-
E-mail: imachado@ufpe.br iting animals towards flowers and/or fruits. We made a new record of the lizard Tro-
pidurus semitaeniatus foraging on flowers and fruits of M. ernestii. During the search
Editor for nectar, T. semitaeniatus contacted the reproductive structures of the flowers and
A. Dafni transported pollen attached to its snout. Nectar production started at 14:00 h, with an
average volume of 24.4 ll and an average concentration of solutes of 33%. Approxi-
Received: 9 November 2012; Accepted: 20 mately 80% of the seeds of M. ernestii found in the faeces of T. semitaeniatus germi-
May 2013
nated under natural conditions. The roles of T. semitaeniatus as pollinator and seed
disperser for M. ernestii show a clear relationship of double mutualism between two
doi:10.1111/plb.12063
endemic species, which may result from the environmental conditions to which both
species are subject. Seasonality, low water availability and arthropod supply in the
environment, high local lizard densities, continuous nectar production by the flower
and fruits with juicy pulp may be influencing the visits and, consequently, pollination
and seed dispersal by lizards in this cactus.

Figueira et al. 1994; Valido & Nogales 1994; P
erez-Mellado &

INTRODUCTION
Angiosperms provide us with countless examples of visual sig-
nals to be perceived and acted upon by animals visiting the
flowers and fruits to obtain rewards and to simultaneously
provide pollination and seed dispersal services (Hansen 2006;
Valido et al. 2011; Cazetta et al. 2012; Newman et al. 2012).
These are two key processes in the reproductive ecology of
most plants (Hansen & Muller 2009).
Saurophily and saurochory are ecological processes where
reptiles mediate pollination and seed dispersal, respectively


(God
ınez-Alvarez 2004; Olesen & Valido 2004; Valido &
Olesen 2007). Lizards are the least common pollinators and
seed dispersers among vertebrates, since most are predators of
small animals and only rarely consume plants (Iverson 1982,
1985). However, for a large number of plant species, lizards are
keystone pollinators and seed dispersers, and play a central role
in the reproductive ecology of plants of different families,
mainly on islands (Olesen & Valido 2003, 2004; Valido &
Olesen 2007).
In their revision, Cooper & Vitt (2002) highlight that out of
ca. 4.000 lizard species, only ca. 1% are herbivores. However,
several analyses show that the mutualism between lizards and
plants has been underestimated (Whitaker 1987; Traveset 1993;
Casas 1997; Traveset & Sa
ez 1997; Olsson et al. 2000; P
erez-
Mellado et al. 2000; Nyhagen et al. 2001; Olesen & Valido
2003, 2004; Sazima et al. 2005; Rodr
ıguez-Rodr
ıguez & Valido
2008; Casado & Soriano 2010; Fonseca et al. 2012; Ortega-
Olivencia et al. 2012; Rodr
ıguez-Rodr
ıguez et al. 2013), and
that lizards may represent one of the oldest biotic pollen
vectors (Williams & Adam 2010). Most records of lizards visit-
ing flowers or eating fleshy fruits, however, remain restricted to
oceanic islands, which may be affected by factors such as
isolation, aridity and prey availability (Cooper & Vitt 2002;
Olesen & Valido 2003), and only a few records are known for
continents (Olesen & Valido 2003; Williams & Adam 2010).
The majority of interactions of mutual dependence between
animals and plants are generalist, frequently involving dozens
of species; highly specific, one-to-one, mutualistic interactions
are the exception (Jordano 1987, 2010). There are few cases of
a given animal species acting both as a pollinator and seed
disperser in a specialised mutualism, however, for most plants,
the pollinator and seed disperser fauna are different from each
other (Proctor et al. 1996). These cases of double mutualism
are more frequent in habitats poor in animal diversity, such as
deserts, islands, mountains and cliffs, in which plants may use
the few available resources, and consequently, evolve towards

Plant Biology © 2013 German Botanical Society and The Royal Botanical Society of the Netherlands 1
Pollination and seed dispersal by a lizard in a continental ecosystem Gomes, Quirino & Machado

double mutualism, i.e. to use the same animal as both pollina-
tor and seed disperser (Garc
ıa et al. 2012). Examples of double
mutualism by the same vertebrate or invertebrate animal spe-
cies highlight lizards, birds, flying foxes or ants acting as double
mutualists of plants (Ladley & Kelly 1996; Soriano & Ruiz
2002; Kelly et al. 2004; Nyhagen et al. 2005; Hansen & Muller
2009; Rodr
ıguez-Rodr
ıguez & Valido 2011; Garc
ıa et al. 2012).
Cases of double mutualism are extremely rare, with relatively
few examples of plants being pollinated and having their seeds
dispersed by the same animal species. A good example of this,
combining both lizard pollination and lizard seed dispersal, is
found on the island of Mauritius, where the gecko Phelsuma
cepediana is currently the sole pollinator and seed disperser of
the critically endangered endemic plant Roussea simplex (Rous-
seaceae) (Hansen & Muller 2009). Other examples are some
mistletoe species in New Zealand (by bellbirds; Kelly et al.
2004), three species of columnar cacti in South America (by
bats; Soriano & Ruiz 2002), one species of Ebenaceae (Diospy-
ros tessellaria) in Mauritius (by flying fox; Nyhagen et al. 2005)
and the dioecious herb Borderea chouardii (Dioscoreaceae) (by
ants; Garc
ıa et al. 2012).
In the present study, in addition to recording the first case of
pollination by lizards in Cactaceae, we report the first case of
double mutualism in the Caatinga, a Brazilian semiarid ecosys-
tem. We analyse a highly specialised system of pollination and
seed dispersal, and discuss which ecological conditions may
favour this rare type of animal–plant mutualism involving
double specificity.
MATERIAL AND METHODS
Study species and site
Fieldwork was carried out in the private reserve Almas Farm
onio Natural – RPPN – Fazenda
(Reserva Particular do Patrim^
Almas) located within the municipalities of Sum
e and S~ao Jos
e
dos Cordeiros (7°28′15″ S and 36°52′51″ W), state of Para
ıba,
in the semiarid region of northeast Brazil. The Almas Farm is
located in the region having the lowest rainfall record for the
Caatinga. The municipality of S~ao Jos
e dos Cordeiros has an
annual rainfall from 400 to 800 mm, with the rainy period
between December and January. The municipality of Sum
e has
an average annual rainfall of 560  230 mm, concentrated in
February, March and April. In these 3 months rainfall amounts
to ca. 330 mm; this corresponds to ca. 60% of the annual total
and results in a dry season that is frequently longer than
8 months. The climate of the region is characterised by extreme
conditions, with high radiation, low relative humidity and
cloudiness, and high average annual temperature varying from
26 to 33 °C (Prado 2003); it is classified as ‘warm semiarid’
(Bsh) with summer rains, according to the K€ oppen system.
In the study site, we carried out focal studies on one popula-
tion of M. ernestii on rocky outcrops, composed of four
patches of 15 individuals each, 10 m away from each other. We
also used a fourth patch composed of 50 individuals and
located 8 m away from the others, mainly to estimate the daily
production of flowers and fruits.
Melocactus ernestii Vaupel subsp. ernestii is a globose or
cylindrical cactus that occurs exclusively in eastern Brazil, with
disjunct populations in gneissic inselbergs and crystalline and
sandstone rocks surrounded by an arboreal Caatinga matrix,
represented predominantly by xerophilous vegetation (Taylor
& Zappi 2004). M. ernestii shows continuous flowering (Quiri-
no 2006), and there are records of its diurnal flowers being
visited by different species of hummingbirds and butterflies
(Rom~ao et al. 2007), as is common for the genus (see Table 1).
In earlier studies, Britton & Rose (1963), Taylor (1991) and
Rocha & Agra (2002) mentioned lizards and birds as potential
seed dispersers of representatives of the genus Melocactus.
Rom~ao et al. (2007), in their study with M. ernestii, found
lizards (Cnemidophorus sp. and Tropidurus sp.), birds and ants
acting as seed dispersers.

Table 1. Species of Melocactus in different sites around the world, and their recorded pollinators and seed dispersers (hb, nd).

species study site nectar pollinator seed disperser reference

M. andinus Andean xeric 5.3 ll, 22.8% hb, ant, bee + n.d. Nassar et al. (2007)
patches, Venezuela small beetle
M. conoideus Caatinga, Brazil n.d. n.d. ants Brito-Kateivas & Corr^ea (2012)
M. curvispinus Xeric areas, Venezuela 163.1 ll (daily flower 1), hb + bee n.d. Nassar & Ram
ırez (2004)
29.1 and 30.2%
M. ernestii Caatinga, Brazil 32.2 ll* (day 1) hb, butterfly + birds, Rom~ao et al. (2007);*
lizard* ants+ lizards
M. glaucescens Caatinga + Cerrado, Brazil 30.4  3.7 ll, hb + butterfly lizards + ants Colacßo et al. (2006); Fonseca et al. (2012)
29  3%
M. intortus Subtropical dry forest, n.d. hb, ant + bee lizards + ants Fagua & Ackerman (2011)
Puerto Rico
M. paucispinus Caatinga + Cerrado, Brazil 33.6  4.1 ll, hb + butterfly birds + lizards Colacßo et al. (2006); Fonseca et al. (2012)
29  3%
M. salvadorensis Caatinga, Brazil n.d. hb n.d. Raw 1996
M. schatzlii Andean xeric patches, 4.4 ll, 23.3% hb, ant, bee + birds + lizards Nassar et al. (2007); Casado & Soriano (2010)
Venezuela small beetle
M. violaceus Restinga + Caatinga, Brazil n.d. hb lizards Figueira et al. (1994)
M. zehntneri Caatinga, Brazil 35–41  1.18 ll, hb n.d. Locatelli & Machado (1999)
27  0.57%

hb = hummingbird; n.d. = not determined.*This study only.

2 Plant Biology © 2013 German Botanical Society and The Royal Botanical Society of the Netherlands
Gomes, Quirino & Machado
Tropidurus semitaeniatus Spix 1825 (Squamata, Tropiduri-
dae) is a lizard species that is endemic to the Caatinga, a dry
forest of northeast Brazil; it is broadly distributed in rocky
habitats (Passos et al. 2011). It is a medium-size lizard (maxi-
mum snout–vent length = 72.4 mm; Brandt & Navas 2011),
with a generalised diet including invertebrates, leaves, fruits
and seeds (Cooper & Vitt 2002; Ribeiro et al. 2008; Fonseca
et al. 2012). Observations carried out for three species of Tro-
pidurus show these lizards are carnivorous and/or omnivorous,
which suggests geographic variation in their diet and temporal
variation in the consumption of plants (Cooper & Vitt 2002).
Reproductive phenology and floral biology
We carried out monthly phenological monitoring of M. ernestii
from March 2009 to September 2011, by directly counting
flowers and fruits of 15 individuals. We also observed colour,
odour, period of anthesis, concentration and volume of nectar
of the flowers. We measured the size and depth of the corolla
and reproductive structures using a stereomicroscope for 15
flowers of different individuals fixed in 70% alcohol. We
observed other 15 flowers of M. ernestii to record the dynamics
of anthesis, i.e. the time of appearance of flowers (buds) on the
surface of the cephalium, flower opening and end of anthesis.
We measured the volume and concentration of nectar in
recently opened flowers of different individuals (N = 15) at
intervals of 1–4 h (14:00–18:00 h) using microsyringes (50 ll)
and a pocket refractometer (Atago 23–64%). The cephalium
was covered with nylon bags on the previous day to avoid
contact between flowers and animals. Values of total sugar were
transformed into milligram, using a constant table provided in
Galetto & Bernardello (2005).
Voucher specimens of M. ernestii subsp. ernestii were depos-
ited in the Herbarium Lauro Pires Xavier at the Department of
Botany, Federal University of Para
ıba (JPB – 45095).
Visiting animals
Focal observations were made on 45 individuals of M. ernestii
during eight non-consecutive days. Each session of focal obser-
vations lasted ca. 1 h, interpolated with 30-min intervals,
summing up to 78 h of focal observation, being 40 h on the
fruits and 38 h on the flowers. We recorded in the field the
number of visitors, frequency of visits, time of permanence on
the cephalium, resource searched, behaviour towards flowers
or fruits, and the site of contact with reproductive structures.
The occurrence of agonistic behaviour was also documented.
The behaviour of lizards during the visits was photographed and
monitored from 08:00 to 18:00 h, corresponding to the whole
period when flowers and fruits were available. Since M. ernestii
shows continuous flowering and fruit set (Quirino 2006), we
concentrated the sampling of visiting animals in the months of
flowering (September) and fruit set peaks (November).
Exclusion pollination experiments
We used short-term exclusion experiments to investigate the
potential role of Tropidurus semitaeniatus as a pollinator of
M. ernestii. The exclusion experiments were set up on sunny
days with no rain, during the flowering period of M. ernestii
in September 2011, consisting of three treatments: (i) control
Plant Biology © 2013 German Botanical Society and The Royal Botanical Society of the Netherlands
Pollination and seed dispersal by a lizard in a continental ecosystem
– all pollinators having access; (ii) no visitors – self-pollina-
tion; and (iii) only lizards having access. We randomly
selected 20 flowers of ten different plants (two flowers per
individual) for each treatment, prior to flower opening. The
treatments were conducted to compare the fruit set. In the
control (treatment 1), flowers in pre-anthesis were marked
and left free to allow access to all visitors. The flowers in
spontaneous self-pollination (treatment 2) were bagged at
pre-anthesis. To exclude flying pollinators (hummingbirds
and butterflies), but allowing lizard visits, the Melocactus
individuals were covered with a cube of plastic mesh with
2 9 2 mm gauge. After visits of lizards, the cephalium was
covered with nylon bags, excluding both kind of visitor
(flying animals and lizards). After 15 days, we proceeded to
evaluation of fruiting in all three treatments. The chi-square
test was used to compare fruit set, fruit size and number of
seeds per fruit between the treatments.
Seed germination tests
We investigated experimentally the effect of passage of seeds
through the intestine of lizards. The experiment was composed
of two germination treatments: (i) with seeds of the control
group removed directly from fruits; and (ii) with seeds
dispersed by lizards. The seeds of the control group (N = 100)
were obtained from fruits of three different plants chosen at
random, so that each Petri dish had seeds of three plants. To
obtain dispersed seeds, we collected faecal samples of different
lizard individuals (N = 8 scats) at distances of up to 5 m from
the closest reproductive adult cactus. The seeds found
(N = 102) were submitted to germination tests under natural
conditions in a greenhouse (average temperature = 28 °C). We
repeated each treatment four times, using 25 seeds placed on a
Petri dish, covered with two layers of filter paper moistened
with distilled water. The dishes were sealed with transparent
masking tape and no contamination by fungi was observed.
We counted the seeds that germinated and removed them from
the Petri dish each day for 30 days; germination was defined as
the time when the radicle tip emerged ≥1 mm from the seed
coat (Meiado 2012). For each seed germination treatment, we
calculated the germinability. Differences in germination
percentage of dispersed seeds with the germination percentage
of seeds collected directly from fruits (control) were tested for
statistical significance using a Mann–Witney U-test in the
program STATISTICA 10.0 (P < 0.05; Statistica Inc., Tulsa,
OK, USA).
RESULTS
Reproductive phenology and floral biology
Melocactus ernestii produced flowers and ripe fruits in
all months of observation (Fig. 1); the flowering peak occurred
in February and September, and April and November had
the highest fruit-set. The analysed flowers were tubular
(12.5  2.5 mm long 9 2.0  1.5 mm wide), lilac, with
20  25 tepals (4.0  2.0 mm long 9 1.0  1.0 mm wide)
and had stamens (ca. 200) of different sizes (2.0  1.0 mm
long), which surrounded the style (8.0  1.5 mm long), the
lobed stigma (4–5 lobules) and the nectariferous chamber
located at the base of the flower.
3
Pollination and seed dispersal by a lizard in a continental ecosystem
Fig. 1. Flowering (□) and fruit set (■) of Melocactus ernestii subsp. ernestii
(N = 15) and rainfall in a rocky outcrop area of the private reserve Almas
Farm (Reserva Particular do Patrimo ^nio Natural RPPN Fazenda Almas), state
of Para
ıba, northeast Brazil, from March 2009 to September 2011.
Flower buds remained inside the cephalium and appeared
gradually on its surface, at around 11:00 h on the day of anthe-
sis, which began at 14:00 h and lasted until 18:00 h. The pollen
was already available ca. 2 h before the beginning of anthesis,
whereas nectar started to be produced at the beginning of
anthesis, with an average volume of 10.1  2.08 ll (N = 15) in
the first hour of observation; there was a peak in production,
doubling the volume of nectar from 16:00 h onward. The vol-
ume of accumulated nectar was 32.2 ll (N = 11) and the
average concentration of solutes was 33  5.5%, corresponding
to an average of 9.39  2.17 mg sugar per flower. The fruits
were conical and bright lilac in colour, with seed heads
of 21.32 9 6.78 mm, average 56  5 black seeds, each of
1.04 9 1.01 mm immersed in the aqueous transparent pulp
of the seed head.
Floral and fruit visits
We monitored visits to flowers (N = 29 records in 38 h;
0.76 visit
h 1) and fruit consumption (N = 41 records in
40 h; 1.02 visit
h 1) of M. ernestii by the lizard T. semitaenia-
tus. Each day, the lizards performed an average of 4  2
visits to flowers and 3  1 visits to fruits; or a whole day,
the number of visits of T. semitaeniatus ranged between one
to six and one to four, respectively, to flowers and fruits. On
the flowers, we also observed visiting hummingbirds and
butterflies, but at a lower frequency (N = 6 and 5 records,
respectively). The visits of T. semitaeniatus to the flowers
started at 14:00 h (Fig. 2), coinciding with the beginning of
anthesis, and reached a higher frequency of visits around
15:30 h, a time of day with very high temperatures (around
33 °C). Visits of hummingbirds also occurred during the
whole period of anthesis, and the last visit was recorded at
17:30 h. Lizards and hummingbirds took turns in visits to
flowers of M. ernestii, without an exclusion of one group by
the other. However, we observed agonistic interactions
between adult and juvenile individuals of T. semitaeniatus,
and between individuals of T. semitaeniatus and humming-
birds. Lizards stretched their body and acquired an erect
position on the cephalium, not allowing other lizards and
4 Plant Biology © 2013 German Botanical Society and The Royal Botanical Society of the Netherlands
Gomes, Quirino & Machado
Fig. 2. Temporal distribution of visits of Tropidurus semitaeniatus to fruits
and flowers of Melocactus ernestii subsp. ernestii during the day in the pri-
vate reserve Almas Farm (RPPN Fazenda Almas), state of Para
ıba, northeast
Brazil. Total: 78 h of observations; 42 h from 08:00 to 13:00 h and 36 h
from 13:00 to 18:00 h.
hummingbirds to come close to the same individual of
Melocactus.
In 38 h of focal observations we recorded visits of lizards to
29 flowers, and each visit lasted 12  5 s. When a T. semitaeni-
atus lizard identified an open flower, it climbed the cephalium,
inserted the head repeatedly into the flower (Fig. 3A and B),
and then moved to other flowers of the same or another cepha-
lium, visiting one to two flowers on the same cephalium.
During the visits, T. semitaeniatus contacted the anthers and
stigma; we observed pollen grains attached to its snout after
each visit. At the beginning of anthesis, the ends of the inner
tepals moved apart, forming a small space that progressively
increased in diameter until the complete opening of the flower
(Fig. 3C), reaching 3.5 mm when the flowers were completely
open. At that moment, the contact with reproductive struc-
tures occurred more frequently, as it was easier for the lizards
to access the stigma that is located beyond the anthers.
Considering that each patch of M. ernestii studied in insel-
bergs was composed of 15 reproductive individuals, with an
average production of three flowers a day (from one to five
flowers per individual), there were on average 45 flowers avail-
able for visitors each day, which would represent a total of
422.55 mg sugar
day 1 available for pollinators. Out of these
45 daily available flowers, we recorded a maximum of six
flowers visited by lizards during one full day, which equates to
consumption of 56.34 mg sugar
day 1; other pollinators
(hummingbirds and butterflies) consumed the remaining
available nectar.
In parallel, we also recorded 12 events of florivory (Fig. 3D),
in which lizards consumed the succulent petals. We observed
florivory events after the visits of lizards to consume nectar.
Possible damage caused by the consumption of petals, in terms
of loss of flower attractiveness, did not compromise the repro-
ductive success of this species of Melocactus, because lizards
first drank the nectar, and in consecutive visits consumed the
petals. Moreover, as flowers remained buried in the cephalium,
the ovary and ovules were protected from predation even after
the removal of petals and, consequently, the flowers from
which petals were consumed still produce fruits normally
(N = 12).
The visits of lizards to fruits started at 09:00 h, with the
removal of diaspores exposed on the cephalium (more than
50% of fruits on each cephalium), which were easier to remove.
Gomes, Quirino & Machado Pollination and seed dispersal by a lizard in a continental ecosystem

A C

B E

Fig. 3. Tropidurus semitaeniatus on the cephalium of

Melocactus ernestii (A) and with the head inside a flower
(B). Flower of M. ernestii at the beginning of the anthe-
sis, with the corolla intact (C) and with petals eaten (D)
by the lizard. T. semitaeniatus ingesting a fruit of M. ern-
estii (E), highlighting the amount of water in the pulp of
the fruit. All pictures were taken in an area of rocky out-
crops in the private reserve Almas Farm (RPPN Fazenda
Almas), state of Para
ıba, northeast Brazil.

This procedure was very rapid, lasting about 20 s, time enough Table 2. Reproductive success of Melocactus ernestii subsp. ernestii (Cacta-
for the lizard to climb on the cephalium and remove the fruit. ceae), fruit size and number of seeds per fruit in the three treatments (con-
Next, the fruit was completely consumed while the lizard was trol, spontaneous self- and lizard pollination) in the private reserve Almas
still on the cephalium or just after it climbed down the plant. Farm (RPPN Fazenda Almas), state of Para
ıba, northeast Brazil.
The lizards turned the fruit to facilitate ingestion and avoid reproductive
losing the pulp. In parallel, we collected seeds of M. ernestii success% number of
from the dung of T. semitaeniatus (N = 102 seeds), confirming treatment (fruits/flowers) fruit size (mm) seeds/fruit
the consumption of fruits by these animals. We observed that
juvenile lizards performed most visits to the flowers (N = 17 natural pollination 95 (19/20)a 20.4  1.0 9 4.2  2.1a 52  6a
records), while adults made only 12 visits. Similarly, we also (control)
observed that, of 41 visits to fruits, 25 fruits (60%) were taken spontaneous 60 (12/20)b 19. 3  0.5 9 4.1  2.2a 49  6a
by juvenile lizards. self-pollination
lizard pollination 80 (16/20)a 20.1  0.8 9 5.0  1.5a 51  5a

Exclusion pollination experiments Different letters indicate significant differences (P < 0.05).

In all pollination treatments there was fruit formation

(Table 2), however, the flowers had different percentages of
fruit-set. There were significant differences in the percentage of
fruits produced between natural pollination and spontaneous
self-pollination (v² = 8.654, P = 0.003). The flowers exposed
only to lizards showed an increment of 20% on fruit set, and
there were no significant differences between this treatment
and natural pollination (v² = 2.3263, P = 0.1272), when both
pollen vectors associated with the rate of spontaneous self–
pollination were acting. The fruits formed from the three polli-
nation treatments varied in size and number of seeds, but there
were no significant differences in fruit size and number of seeds
per fruit among the pollination treatments (Table 2).
Germination tests
Tropidurus semitaeniatus lizards (Fig. 3E) had a high quantity
of viable seeds of M. ernestii in their faeces, presenting 12
seeds
scat 1, on average. In the germination test, ca. 80%
(N = 81) of seeds of M. ernestii found in lizard faeces germi-
nated under natural conditions. Germination rates differed
significantly: seeds that passed through the digestive tract of
lizards (N = 20  1 seeds germinated) had a significantly
higher germination rate than seeds of the control group
(N = 4  2 seeds germinated; P < 0.0001).
DISCUSSION
In the interaction system described here (involving a cactus
and a lizard), Tropidurus semitaeniatus acts as a double mutual-
ist, pollinating and dispersing seeds of M. ernestii, and contrib-
uting in two ways to the reproductive success of this cactus.
This is one of the very few records of lizards as double mutual-
ists. Due to its continuous pattern of flowering and fruit set

Plant Biology © 2013 German Botanical Society and The Royal Botanical Society of the Netherlands 5
Pollination and seed dispersal by a lizard in a continental ecosystem
(Quirino 2006; Rom~ao et al. 2007), M. ernestii may be consid-
ered a key resource in the studied Caatinga area. This is espe-
cially true in periods of severe drought and food scarcity for
pollinators and dispersers, since nectar, pollen and fruits are
used by several groups of animals, including lizards, to meet
their energy requirements, as observed in the present study.
Moreover, fleshy fruits and floral parts of these cacti may be
not only a source of carbohydrates, but also of water for these
animals (Figueira et al. 1994; Fonseca et al. 2012). The results
obtained from the pollinator exclusion experiment reinforce
the role of T. semitaeniatus as pollinator of M. ernestii. The
higher fruit production (95%) in the natural pollination treat-
ment, and spontaneous self- + lizard pollination (80%), when
compared to spontaneous self-pollination (60%), shows that
the biotic pollen vectors (including lizards, hummingbirds and
butterflies) contribute to increase fruit production. The polli-
nation by lizards increased fruit set by 20%; however, when we
consider all potential pollinators, total fruit set increased by
35%, with hummingbirds contributing 20% and butterflies
15%.
The role of hummingbirds as effective pollinators in the
reproduction of globose cacti is well known among the Melo-
cactus species already studied (e.g. Locatelli & Machado 1999;
Nassar & Ram
ırez 2004; Colacßo et al. 2006; Rom~ao et al. 2007;
Nassar et al. 2007; Fagua & Ackerman 2011; Table 1). However,
the frequency of visits of T. semitaeniatus and the contact with
reproductive structures of M. ernestii flowers make this lizard
species an effective pollinator of this cactus, helping in its
sexual reproduction success. The presence of pollen grains
attached to the snout corroborates the discovery of this lizard
as a pollen vector of the studied cactus. Pollen grains adhere
and are easily transported on the body and scales, mainly on
the ventral surface, snout and throat of this lizard (Williams &
Adam 2010). Indeed, for several plant species lizards are
keystone pollinators and seed dispersers, and play a central role
in the reproductive ecology of plants of different families,
particularly on islands (Olesen & Valido 2003, 2004; Valido &
Olesen 2007).
The antagonistic behaviour observed between adult and
juvenile lizards may interfere with pollen flow among individ-
ual plants, and favour increased geitonogamy in the popula-
tion, since M. ernestii is self-compatible, similar to
M. paucispinus (Colacßo et al. 2006). This antagonistic behav-
iour creates a cephalium monopoly, where the lizard visits
all the flowers in the same cephalium, thus favouring geito-
nogamy. On the other hand, these animals also promote
xenogamy, as they can visit various cephalia because these
small cacti grow close to the ground, which facilitates access
of the lizards to many plant individuals. Moreover, the
opening of the ends of the inner tepals increases the diame-
ter of the flower, which facilitates initial contact of the lizard
with the stigma, which is located beyond the anthers, also
favouring xenogamy. Hence, the territorial behaviour of liz-
ards would tend to keep pollen flow at short distances, most
likely favouring mating events among neighbouring cacti,
especially as these cacti live in vegetation islands known as
inselbergs. Furthermore, when the lizard is over the cephali-
um, it can favour cross-pollination mediated by humming-
birds, since the presence of the lizard scares off these birds,
which visit only one flower and then move to another ceph-
alium.
6 Plant Biology © 2013 German Botanical Society and The Royal Botanical Society of the Netherlands
Gomes, Quirino & Machado
The ornithophilous characteristics (Faegri & Pijl 1979) of
M. ernestii flowers are similar to those of other congeners
with records of pollination by hummingbirds (e.g. Locatelli
& Machado 1999; Nassar & Ram
ırez 2004; Colacßo et al.
2006; Nassar et al. 2007; Fagua & Ackerman 2011; Table 1).
However, the colour of the flowers, which is the same as
that of the fruits, and their location close to the ground
(Fig. 3C) makes them attractive to lizards, as they are easily
identified and accessed by these animals. In fact, we
observed that lizards and hummingbirds are both pollinators
of M. ernestii. These attributes of M. ernestii associated with
the semiarid environment may induce a mixed pollination
system that combines ornithophily and saurophily. A mixed
pollination system was also observed recently in Scrophularia
calliantha (Scrophulariaceae), in which birds, bees and juve-
nile lizards of the species Gallotia stehlini (Lacertidae) share
the pollination services of S. calliantha (previously consid-
ered as exclusively ornithophilous; Ortega-Olivencia et al.
2012) as well as Isoplexis canariensis (Rodr
ıguez-Rodr
ıguez
et al. 2013). Although the flowers of M. ernestii are classified
as typically ornithophilous, with records for other popula-
tions of visits exclusively by hummingbirds (Rom~ao et al.
2007), environmental factors may be inducing visits from
different animal groups, such as lizards. This mixed system,
involving lizards as pollinators, may also occur in other
globular cacti in Caatinga areas, highlighting the need for
further studies on the reproductive system of cacti, as pro-
posed by Guerrero et al. (2011).
Availability of nectar at high concentrations in M. ernestii
may also be attractive to lizards. Scogin (1985), who analysed
nectar variations in 43 species of cacti, observed that among
flowers with different pollinators (hummingbirds, moths and
bats) there are no significant differences in the concentration
of solutes. Nectar concentration of M. ernestii (28–38%) is
higher than recorded in the literature for species pollinated by
hummingbirds, where concentrations vary from 20 to 26%
(Pike & Waser 1981; Stiles & Freeman 1993; Proctor et al.
1996). Although a high nectar concentration is common in
Melocactus, at least among the already studied species
(Table 1), the previous concentrations observed have always
been lower than that found in M. ernestii. Although the role of
reptiles as pollinators of Melocactus or other cacti not yet fully
understood, as lizards have a high energy requirement (Vitt
1995), the high nectar concentration of M. ernestii could
explain why lizards are attracted to its flowers. The flowers of
Melocactus represent a reliable source of food and water for
lizards throughout the year; for the plants, the presence of an
alternative pollinator may contribute to reproductive success.
Hence, the conservation of these two taxa, both endemic to the
Brazilian Caatinga, needs to consider plant–lizard mutualistic
interactions.
Plant–lizard interactions seem to be more broadly distrib-
uted than only on islands, as was frequently assumed (Olesen
& Valido 2003). According to these authors, saurophily and
saurochory predominate in insular ecosystems because lizard
populations reach high densities in such sites, and expand their
diets to include nectar and fruit as an escape from predation
risk. We suggest that this interaction may also occur in other
Melocactus species, or even in other cactus genera and families,
mainly those of very dry environments, such as rocky outcrops
(inselbergs) having scarce resources. The degree of aridity of
Gomes, Quirino & Machado Pollination and seed dispersal by a lizard in a continental ecosystem

the environment and the consequent water deficit are ecologi-
cal factors that may lead to the consumption of plants by
lizards that forage for water and nutrients, increasing the prob-
ability of saurophily and saurochory in the driest areas of the
Caatinga. This has been observed in visits of lizards (Euprepis
atlanticus), in search of water and sugar, to flowers of Erythrina
velutina, an ornithophilous Leguminosae of an oceanic island
in northeast Brazil having a high water deficit (Sazima et al.
2005).
Although this is the first record of double mutualism for
the Caatinga, there may be other cases of double mutualism
in continental areas with extreme environments in the semi-
arid region of northeast Brazil. We suggest that the biogeo-
graphic conditions to which both species are subject (low
water availability and low arthropod supply in the environ-
ment, high local lizard densities), the continuous nectar
production and fleshy (succulent) fruits of this cactus species
may influence both pollination and seed dispersal by lizards.
This highly seasonal ecosystem, with an extreme dry season,
might harbour highly specialised mutualistic interactions, as
reported in the present study. Conditions favouring extreme
cases of double specificity may be very rare in other habitat
types where abiotic conditions do not limit generalised
mutualisms. Hence, it is also possible that over larger areas
and in other localities with milder environmental conditions,
there may be broader geographic mosaics of interactions (e.g.
Thompson 2005) involving this Melocactus species and its
pollinators and dispersers, where either (or both) of the
partners could interact with multiple species. We suggest,
however, that extreme abiotic conditions may favour the
appearance of highly specialised mutualisms, involving strong
dependence on a single vector species for both pollination
and seed dispersal.
ACKNOWLEDGEMENTS
The management of Almas Farm and Joel Queiroz provided
fieldwork logistics. Dr. Nigel Taylor, Dr. Marcos Vin
ıcius
Meiado, and Washington Luiz Vieira provided general infor-
mation on the species. Dr. Pedro Jordano and Dr. Alfredo Va-
lido read the manuscript and made invaluable suggestions. The
Brazilian Research Council (CNPq) granted an undergraduate
scholarship through the project ‘PELD/Caatinga: Estrutura e
Funcionamento’, and later Capes granted VGNG a Master’s
scholarship; CNPq for also provided a research productivity
fellowship to ICM.

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