South African Journal of Botany 108 (2017) 346–351

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South African Journal of Botany

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Multiple shoots of Carapa surinamensis seeds: Characterization and

consequences in light of post-germination manipulation by rodents
D.N. de Souza Ferreira a,⁎, J.L.C. Camargo b, I.D.K. Ferraz a
a
Brazilian National Institute for Amazonian Research (INPA), Av. André Araújo, 2936, Manaus, AM, Brazil
b
Biological Dynamics of Forest Fragments Project (BDFFP–INPA/STRI), Av. André Araújo, 2936, Manaus, AM, Brazil

a r t i c l e i n f o a b s t r a c t

Article history: Post-germination manipulation of seeds by rodents arrests shoot development, yet seed tissues may remain alive
Received 3 May 2016 becoming an important food supply over several months. Seeds of Carapa surinamensis show polyembryony in
Received in revised form 1 August 2016 the Central Amazon, which could counter predatory seed manipulation. To test this concept, we characterized
Accepted 23 August 2016
the polyembryony of C. surinamensis seeds by assessing the number of shoots per seed over time and with respect
Available online 6 September 2016
to seed mass. We also simulated rodent manipulation over two post-germination stages, comparing re-sprouting
Edited by AK Cowan of C. surinamensis with Carapa guianensis, the latter usually producing only one shoot. Metabolic activity of the
seed reserves was tested by tetrazolium staining. We found multiple shoots in over 50% of C. surinamensis
Keywords: seeds and up to five shoots per seed; however, the number of shoots was not related to seed mass. The interval
Crabwood between successive shoot production by the same seed was up to 46 days. Some re-sprouting occurred after
Herbivory radicle pruning (45% of C. surinamensis and 57% of C. guianensis seeds). The removal of two meristems resulted
Carapa in “zombie seeds” (76% in C. surinamensis and 81% in C. guianensis). Polyembryony did not provide an advantage
Polyembryony in re-sprouting in comparison to monoembryonic seeds. Seed manipulation maintained the viability of seed
reserves under moist and hot (25 °C) conditions for 6 months.
© 2016 SAAB. Published by Elsevier B.V. All rights reserved.

1. Introduction
Plant–animal interactions, especially herbivory (consumption of
seeds and vegetative parts), contribute to the maintenance of tree
diversity in Neotropical forests, as they affect plant recruitment and dis-
tribution (DeMattia et al., 2004; Knight, 2004). Such interactions may
have positive or negative effects, depending on the species
involved, developmental stage, and the affected organs, as well as toler-
ance or resistance of the plants to damage (Davidson, 1993; Begon et al.,
2006).
Some evolutionary plant strategies have been proposed to minimize
loss caused by seed consumption and herbivory (Davidson, 1993). For
example, rapid germination escaping consumption and rapid depletion
of seed reserves by the seedling. Alternatively, delayed germination
until the next period of food abundance, when rodents have abandoned
old hoarding sites (Jansen and Forget, 2001). Several seeds in a single
dispersal unit may be another strategy because diaspores with multiple
seeds have more than one chance to germinate, even after intervention
by vertebrates and insects (Bradford and Smith, 1977; Garrisson and
Augspurger, 1983; McEuen and Steele, 2005). Similarly, polyembryony
⁎ Corresponding author.
E-mail addresses: denajara@gmail.com (D.N. de Souza Ferreira), camargo@inpa.gov.br
(J.L.C. Camargo), iferraz@inpa.gov.br (I.D.K. Ferraz).
(more than one embryo in a single seed) may provide the same advan-
tage as dispersal units with multiple seeds.
Polyembryony can result from the division of the zygote, but also by
adventitious embryogenesis, for example, of nucellus cells. Depending
on the origin, the embryos can be genetically different (Ganeshaiah
et al., 1991; Bewley et al., 2013). Little is known about the outcome of
polyembryony, which seems to increase the fitness of the mother
plant by increasing the chances of establishment of at least one of the
seedlings from the same seed (bet hedging) (Ganeshaiah et al., 1991;
Ladd and Cappuccino, 2005; Blanchard et al., 2010). Polyembryony
can also be beneficial, as more seedlings are produced by the same
investment of the mother plant (Uma-Shaankeer and Ganeshaiah,
1997). On the other hand, embryos in the same seed share reserves
(e.g., cotyledons, endosperm), which may cause competition during
germination and establishment, consequently reducing vigor. Competi-
tion may be caused by external factors because the descendants require
similar environmental conditions (Cheplick, 1992; Mendes-Rodrigues
et al., 2012).
Polyembryony was found in 115 of 348 plant families and adventi-
tious embryony in 52 families (see Carman, 1997). In Meliaceae, poly-
embryony appears to be an unusual phenomenon and was described
only for Melia L. and Azadirachta A. Juss. while multiple shoots or poly-
embryonic seeds were reported in Amoora rohituka (Roxb.) Wight &
Arn. (synonym Aphanamixis polystachya) and Carapa surinamensis

http://dx.doi.org/10.1016/j.sajb.2016.08.015
0254-6299/© 2016 SAAB. Published by Elsevier B.V. All rights reserved.
 D.N. de Souza Ferreira et al. / South African Journal of Botany 108 (2017) 346–351
Miq. (formerly Carapa procera D. C.) (Gosh, 1972; Fisch et al., 1995;
Wilde, 2007; Rai, 2014; Hiwale, 2015).
Carapa is a geographically widespread genus occurring in tropical
forests of the Americas, as well as Africa and India (Pennington et al.,
1981). Since 1878, several revisions had been published for Carapa
(De Candolle, 1878; Harms, 1940; Staner, 1941; Noamesi, 1958;
Pennington et al., 1981; Styles, 1981; Kenfack, 2011; Scotti-
Saintagne et al., 2013), variously reducing or increasing the number
of species in the genus, but a resolved taxonomy of the genus is still
under debate.
Animals that scatter hoard seeds for latter consumption are found
in several ecosystems and continents (Fox, 1982; McEuen and Steele,
2005; Jansen et al., 2006; Cao et al., 2011; Rusch et al., 2014; and
Zhang et al., 2014). Some scatter-hoarding animals manipulate seeds
prior to hoarding, which in many cases does increase the longevity of
the food source. A post-germination manipulation, to reduce or prevent
germination, was observed in tropical rainforest of French Guiana,
where red agoutis (Myoprocta exilis, Dasyproctidae) pruned the emerg-
ing sprouts of C. procera (Meliaceae). The seeds did not re-sprout after
the manipulation; however, the tissues remained alive and the reserves
did not decay, creating “zombie seeds” as a long-term food source for
the animals (Jansen et al., 2006).
Likewise, animals perform seed manipulation before germination in
white oak (Quercus alba L. Fagaceae), where the seeds are bitten in the
embryonic axis region by the gray squirrel (Sciurus carolinensis) in
North America (Fox, 1982). A similar behavior was observed with squir-
rels and seeds of Quercus variabilis Blume and Castanea henryi (Skan)
Rehder & E.H. Wilson (Fagaceae) in China (Xiao et al., 2009). Four
other rodents (Niviventer confucianus, N. fulvescens, Rattus flavipectus,
and Maxomys surifer) intervened in the germination process of
Pittosporopsis kerrii Craib (Icacinaceae) (Cao et al., 2011).
In the Manaus region of the central Amazon, two species of Carapa
occur, C. guianensis Aubl. and C. surinamensis (previously identified as
C. procera, see Kenfack, 2011; Scotti-Saintagne et al., 2013). An impor-
tant and common characteristic of the genus is that a single reserve
tissue is formed by the fused cotyledons, in which a tiny embryonic
axis is inserted (Harshberger, 1902). Carapa guianensis seeds have one
embryonic axis; however, C. surinamensis may have several embryonic
axes inserted in the reserve tissue and may produce multiple shoots,
each connected by two petioles to the same seed reserve (Fisch et al.,
1995; Amoêdo, 2015). Multiple shoots are not known for any other
species of the genus.
Fig. 1. Seed collection sites in Central Amazon, Brazil.
347
The objective of our research was to explore the possible advantages
and disadvantages of multiple-shooted seeds of C. surinamensis with
the monoembryonic seeds of C. guianensis, in light of rodent post-
germination seed manipulation, survivorship, time period between ger-
mination events, the relationship to seed mass and of individual shoots
from the same seed.
2. Materials and methods
2.1. Tree species and seed characteristics
The two species of this study are naturally found along seasonally
flooded forests (locally named várzea and igapó) as well as in terra
firme forests. C. guianensis, has a wider geographical distribution, ex-
tending from Central America through Chocó, Colombia, and including
the entire Amazon Basin. By contrast, C. surinamensis is geographically
restricted to the equatorial region (Scotti-Saintagne et al., 2013).
Based on voucher specimens deposited at the herbarium of the National
Brazilian Institute for Amazonian Research (INPA) and the description
provided by Kenfack (2011), we assume that the correct name for
C. procera seeds collected near Manaus as cited in earlier studies (Fisch
et al., 1995; Ferraz et al., 2002) is C. surinamensis Miq. and we refer to
the species under this name hereafter. The fruits of both species are
large capsules containing up to 20 oil-rich seeds. Seed size varies from
1 to 40 g in C. surinamensis and 1 to 70 g in C. guianensis (Ferraz et al.,
2002). Both have hypogeal and cryptocotylar germination (Fisch et al.,
1995).
2.2. Collection and processing
Seeds used for research were derived from four collection sites in the
state of Amazonas, Brazil: RFAD: Adolpho Ducke Forest Reserve (02°55′
S, 59°59′W); EEST: Experimental Station for Tropical Silviculture
(02°35′55.5″S, 60°02′14.8″W), both areas managed by INPA; Ranch 1:
a private rural property (02°45′19″S, 59°55′58″W) in the municipality
of Manaus; and Ranch 2: a second private rural property (02°47′32.2″
S, 58°24′21.9″W) in the municipality of Silves (Fig. 1).
We collected recently dropped fruits from below parent trees, and
because of the seeds recalcitrant nature (Connor et al., 1998), we
moved them immediately to the laboratory in plastic bags. Seed extrac-
tion followed manual removal of the fruit valves. Until starting the
experimental procedures, seeds were stored in plastic bags at 15 °C,
348 D.N. de Souza Ferreira et al. / South African Journal of Botany 108 (2017) 346–351
Fig. 2. Frequency (%) of polyembryony in Carapa surinamensis assessed by number of
shoots per seed. Mean and standard deviation (n = 249 seeds).
up to a maximum of 7 days. Before germination trials, the seeds were
submerged in water for 24 h to drown potential larvae of Hypsipyla
sp., a lepidopteran known as “seed borer” (Ferraz et al., 2002).
2.3. Characterization of multiple-shooted seeds in C. surinamensis
Weight was determined for each seed prior to germination trials.
Seeds were then sown in plastic boxes (55 × 25 × 15 cm) at 2 cm
depth in moistened vermiculite of medium granulation (Brasil
Minérios®), employing three replicates of 100 seeds each. The boxes
were placed in a greenhouse, roofed with transparent polyethylene
plates. During the experiment, the diurnal temperature varied between
20 and 35 °C, the average relative humidity was 80%, and light incidence
varied between 0.12 and 3.53 × 10 μmol·m−2·s−1. We assessed germi-
nation daily and recorded the date of emergence of each seedling. After
60 days, the viability of ungerminated seeds was verified by a cutting
test (Poulsen et al., 1998), and multiple shoots were separated from
each other and transferred to individual planting pots.
2.4. Simulated seed manipulation by rodents
Pruning of C. surinamensis and C. guianensis at two stages of seedling
development was compared with non-manipulated seeds. For the con-
trol and each manipulation level, we used replicates of 100 seeds. Seed
Fig. 3. Multiple shoots of Carapa surinamensis. (A) Seed with two germinated shoots, (B) Seed with four germinated shoots and (C) Complete individual shoots separated from seed. It is
possible to see that every shoot had complete leaves, an elongated epicotyl and a primary root.
germination was assessed under constant temperature (25 ± 2 °C) and
photoperiod of 12 h (white fluorescent lamps and light intensity
between 0.14 and 0.22 × 10 μmol·m−2·s−1). Seeds were sown above
moistened vermiculite (quality as mentioned above) in plastic trays
(30 × 22 × 7 cm) covered with thin transparent plastic bags to avoid
desiccation. The trial was evaluated daily, and seeds were pruned with
a knife and replaced in the same environmental conditions when they
achieved the desired stage of development: (i) after the protrusion of
the radicle with a length of approximately 5 mm (removal of one
meristem) and (ii) after epicotyl elongation with a height of approxi-
mately 5 cm, for which we pruned the entire shoot (removal of two
meristems). When the seeds resprouted and the epicotyl had reached
a height of at least 5 cm, they were transferred to larger trays in the
greenhouse, as described above and assessed for another 15 days.
Germination was considered recovered if all structures for normal es-
tablishment and at least two photosynthetically active leaves were
developed.
2.5. Long-term seed reserve availability in “zombie seeds”
Seeds with no re-sprouting after pruning remained for 180 days
under controlled conditions as described above. The metabolic activity
of the seed tissues was assessed by staining with 2,3,5-triphenyl
tetrazolium chloride (tetrazolium), where metabolically active tis-
sues were colored from intense pink to red, and non-viable tissues
remained uncolored (Leist and Krämer, 2011). After a longitudinal
cut, the seeds were submerged for 3 h in a tetrazolium solution
(0.1%), at 30 ± 2 °C in the dark, based on a protocol developed for
these species (Amoêdo, 2015). Seeds were considered adequate for
animal consumption when at least 50% of the storage tissues were
pigmented.
2.6. Statistical analysis
We assessed polyembryony using a descriptive analysis of the re-
sults, based on percentage, mean, and standard deviation of the number
of shoots developed by seeds. We assessed the relationship between
seed mass and number of shoots with a one-way ANOVA. For polyem-
bryonic seeds, we calculated the period of time between germination
events of the same seed and the frequency of time intervals between
the first and last germination event of the same seed. In the experiment
of simulated seed manipulation by rodents, a Fisher exact test was
applied to compare non-manipulated seeds with manipulated seeds
 D.N. de Souza Ferreira et al. / South African Journal of Botany 108 (2017) 346–351 349

after pruning at two stages of seedling development and to compare the

two species of Carapa.

3. Results

3.1. Characterization of multiple-shooted seeds in C. surinamensis

C. surinamensis seeds showed 83% (n = 249) germination, and up to

five shoots were developed per seed. Among germinated seeds, 33%
showed two, 13% three, 3% four, and 1% five shoots; therefore, approxi-
mately 50% of the seeds developed multiple shoots (Fig. 2). The number
probably underestimates the true degree of polyembryony since some
of the embryos may have failed to emerge. The shoots were complete,
as all structures necessary for normal development were present (Fig.
3); however, some shoots did not produce chlorophyll. The average
seed mass was 16.7 g (standard deviation = 6.1 g). The smallest germi-
nated seed weighted 1.65 g and had two shoots whereas the largest
seed weighed 39.5 g and had only one shoot. There was no relationship
between seed mass and the number of developed shoots (F = 0.0244,
CV% = 36.77; Fig. 4). In polyembryonic seeds, the first and last germina-
tion from the same seed occurred from 0 to 46 days. Most of the seeds
(72%), produced first and last germination event within the first
10 days after sowing (Fig. 5). This time interval for seeds with just two
shoots was 0 to 34 days, in seeds with three shoots 1 to 46 days, and
in seeds with four shoots 12 to 39 days (Fig. 6). After transplant,
all shoots survived for more than 6 months and showed normal
development.
3.2. Simulated seed manipulation by rodents on C. surinamensis and
C. guianensis
Among the non-manipulated seeds (control), normal shoots were
obtained in 88% of C. surinamensis and 93% of C. guianensis without
any statistical difference between results for the two species (p = 0.33).
After pruning the radicle, re-sprouting was 45% in C. surinamensis
and 57% in C. guianensis. In both species, no re-sprouting was observed
when both root and apical meristem were removed (Fig. 7). Pruning of
the radicle increased the number of dead seeds 1.5 times and doubled it
after pruning of two meristems in C. surinamensis. In C. guianensis, the
consequence of radicle pruning was more pronounced, as seed mortal-
ity increased three to four times (Fig. 7). Only after pruning did seeds
turn into “zombie seeds”. These non-re-sprouting seeds did not deteri-
orate even 6 months after manipulation, keeping reserve tissues alive
(Fig. 8). After only radicle pruning “zombie seeds” were produced in
36% of C. surinamensis and 16% of C. guianensis seeds, increasing to 76%
in C. surinamensis and 81% in C. guianensis after pruning of both radical
and apical meristem (Fig. 7). For both species, the pruning significantly
Fig. 5. Frequency (n) of the time interval (days) between first and last germination of the
same seed in Carapa surinamensis (n = 127 seeds).
reduced shoot production in comparison to the control (p b 2.2e−16).
Re-sprouting after radicle pruning was higher in C. guianensis (p =
0.0049); however, if two meristems were removed, there was no
difference between the species. The development of scar tissue (callus)
was observed in both species at the cutting site (Fig. 8).
4. Discussion
Polyembryony, or seeds producing multiple shoots, were document-
ed for C. surinamensis (Fisch et al., 1995; Amoêdo, 2015); however, they
were never quantified or described in detail, nor mentioned in the tax-
onomic revisions of Carapa. The number of embryos in C. surinamensis
can be high, as we obtained two to five shoots in more than 50% of the
seeds assessed. Each shoot had a primary root and was connected by
two cotyledon petioles to the seed reserve. Eriotheca pubescens (Mart.
& Zucc.) Schott & Endl. (Malvaceae) can produce up to three embryos
(Mendes-Rodrigues et al., 2011), Vincetoxicum rossicum (Kleopov)
Barbar. (Asclepiadaceae) up to four (Ladd and Cappuccino, 2005), and
up to eight embryos were found in seeds of Handroanthus serratifolius
(Vahl) S.O. Grose and H. ochraceus (Cham.) Mattos (Bignoniaceae)
(Mendes-Rodrigues et al., 2012). As multiple embryos compete for
seed reserves (Hotchkiss et al., 2008; Mendes-Rodrigues et al., 2011,
2012) we expected heavier seeds, suggesting a larger amount of re-
serve, to be correlated with higher numbers of shoots. However, we
found no relationship between seed mass and number of seedlings
per seed, suggesting a conservative resource allocation. In fact, the
lightest seed produced more than one seedling whereas the heaviest
seed produced only one. Seedling survival for more than 6 months

Fig. 4. Lack of relationship between seed mass (g) and number of shoots per seed of Carapa Fig. 6. Number of days between first and last germination of polyembryonic seeds
surinamensis. Average and standard deviation (F = 0.0244, CV% = 36.77; n = 127 seeds). according to the number of shoots per seed.
350 D.N. de Souza Ferreira et al. / South African Journal of Botany 108 (2017) 346–351
Fig. 7. Shoot development, seed tissue viability (tetrazolium staining) and dead seeds
6 months after seed manipulation of Carapa surinamensis and C. guianensis by pruning at
two developmental stages after germination: (i) after radicle protrusion with a length of
≥5 mm (pruning of one meristem) (ii) after shoot development with an epicotyl length
of ≥5 cm (pruning of two meristems) in comparison to (o) unpruned seeds (control).
after transplanting indicated that multiple shoots of C. surinamensis
have the potential to develop into an adult individual. This is not true
for all species with polyembryony or multiple shoots. In Amoora
rohituka, only the first and second germinated seedlings can survive
and the others (up to five) die within 30 days (Rai, 2014). In polyembry-
onic seeds of Handroanthus sp., seedling growth was reduced and
mortality increased in a few days after germination, probably due to
competition for seed resources (Mendes-Rodrigues et al., 2012).
Polyembryony may be stimulated by biotic and/or abiotic factors
(Batygina and Vinogradova, 2007). The proportion of mono- and poly-
embryonic seeds may vary within the same population of a species.
For example, in Eriotheca pubescens, a Brazilian tree of savannah-like
forest, the seeds are usually polyembryonic, although some individuals
produce exclusively monoembryonic seeds (Mendes-Rodrigues et al.,
2005). Therefore, it is possible that the populations of C. surinamensis
in Central Amazon vary in embryo number and survival characteristics.
The time between the first and last germination event of the same
seed was up to 46 days, longer than the time span found, for example,
in Vincetoxicum rossicum, which varied between 3 and 20 days (Ladd
and Cappuccino, 2005; Blanchard et al., 2010). We hypothesized that a
long time span may allow a second germination after pruning of the
first seedling and may increase the fitness of the mother plant.
The advantage to more than one embryo germinating from a single
dispersal unit was observed in several species. In Quercus alba, rodent
Fig. 8. “Zombie seeds” 6 months after pruning of Carapa. surinamensis. The growth of a callus is visible at the spot where the radicle or the shoot was cut off (pruned). External view (A).
After transversal cut, reserve tissues with metabolic activity were stained by tetrazolium from intense pink to red (B), dead tissues remained uncolored (C).
handling affected germination and only atypical fruits with multiple
seeds could successfully form seedlings (McEuen and Steele, 2005). Sev-
eral seeds in a single dispersal unit were also observed in Scheelea sp.
(Arecaceae), and the probability of seedling establishment after damage
by rodents or bruchid beetles was proportional to the number of seeds
per fruit (Bradford and Smith, 1977). By contrast, our results suggest
that polyembryony in C. surinamensis confers no advantage over the
monoembryonic seeds of C. guianensis following post-germination ma-
nipulation. The formation of a callus on the spot of pruning, mentioned
for C. procera by Jansen et al. (2006), was confirmed in this study for
both species. The callus may have inhibited the second embryo from
achieving root protrusion in C. surinamensis, as we have observed
successful germination of additional embryos after the death of one
shoot, in non-pruned seeds (Ferreira, 2015).
In Q. mongolica Fisch. ex Ledeb., the capacity to tolerate damage was
attributed to the length of the embryonic axis, which is longer in this
species than in others of the genus; the apical meristem being further
inside the seed is well protected and suffers less damage from handling
(Zhang et al., 2014). The embryonic axes in C. surinamensis are inserted
in the cotyledon tissue and some are deeper inside the seed than others
(Amoêdo, 2015); even so, we observed no recovery from damage.
In temperate and subtropical climates, seeds are buried in autumn
and low winter temperatures inhibit or delay germination and seedling
development. However, in the humid tropics, where ambient tempera-
ture is always favorable to plant growth, non-germinated, and non-
dormant seeds can lose viability in a short time as their organic matter
decomposes rapidly in the tropics. Even so, some animals may manipu-
late seeds before burying to prevent or delay germination. In Aesculus
californica (Spach) Nutt. (Sapindaceae), Quercus alba, C. henryi, and
Q. variabilis (Fagaceae) seed, manipulation by rodents (or by simula-
tion) increased seed death in relation to intact seeds (Fox, 1982;
Mendonza and Dirzo, 2009; Xiao et al., 2009). Simulation of the behav-
ior of four rodent species on seeds of P. kerrii (Icacinaceae) reduced root
formation and seedling establishment; however, 39% of the seedlings
still established via the regeneration capacity of the cotyledons (Cao
et al., 2011).
Seed manipulation after germination was recorded for the first time
in the tropical rainforest of French Guiana (Jansen et al., 2006). The au-
thors simulated animal manipulation and noted re-sprouting in 60% of
the seeds after radicle pruning, whereas pruning at a more advanced
stage reduced recovery to only 20%. Seed manipulation in the present
study resulted in a partial recovery if the radicle was removed in both
species (45% C. surinamensis and 57% C. guianensis); however, no re-
sprouting was seen after eliminating both root and shoot meristems.
In seeds without re-sprouting, Jansen et al. (2006) noticed what they
called “zombie seeds” (i.e., alive tissues, incapable of germination).
Recalcitrant seeds do not form a seed bank, and depletion of reserves
is rapid following germination. When seeds are not maintained under
adequate conditions, they deteriorate quite fast. The period of time
 D.N. de Souza Ferreira et al. / South African Journal of Botany 108 (2017) 346–351
that manipulated seeds remain preserved is known for a few species:
three months in Quercus alba (Fox, 1982) and 5 months for C. henryi
and Q. variabilis (Xiao et al., 2009) or just 5 weeks under tropical condi-
tions for P. kerrii (Cao et al., 2011). In our study, the “zombie seeds” of
both species of Carapa, maintained the viability of seed reserves under
moist and hot (25 °C) conditions over at least 6 months (76% in
C. surinamensis; 81% in C. guianensis), a remarkable phenomenon in
the tropics. The benefit is mutual, the animals maintain oil-rich seeds
that are edible for long periods and the plant, if some of the seeds can
re-sprout, may extend its dispersal area and the time for establishment.
The plant's capacity to form “zombie seeds” may explain part of the
behavior of the rodents in hoarding these seeds instead of immediate
consumption (Jansen et al., 2006). Thus, animal manipulation may be
an effective strategy for food supply even in the Amazonian tropical
rainforest.
Multiple shoots are common in C. surinamensis seeds of the Manaus
region, occurring in over 50% of tested seeds, with up to five shoots per
seed. The number of shoots is independent of seed size. The time inter-
val for germination events of a single seed is up to 46 days, which would
allow for subsequent germination after manipulation. Hence re-
sprouting was only possible after radicle pruning of the first germination
event (45% of C. surinamensis and 57% of C. guianensis seeds). Manipula-
tion at a more advanced stage, by removing two meristems resulted in
“zombie seeds” (76% in C. surinamensis and 81% in C. guianensis). We
conclude that polyembryony does not provide an advantage in re-
sprouting in comparison to monoembryonic seeds in this genus, but
seed manipulation maintains the viability of seed reserves and conse-
quently seed resources under moist and hot (25 °C) conditions at least
for 6 months.
Acknowledgments
This work was funded by FAPEAM (Fundação de Amparo à Pesquisa
do Estado do Amazonas—004/2009 REDEBIO and received additional sup-
port from 015/2014 PAPAC). This work is part of D.N.S.F. MSc thesis in
Ecology Program at INPA, supported by CAPES (Coordenação de
Aperfeiçoamento de Pessoal de Nível Superior). I.D.K.F. is a research
fellow of CNPq.
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