Selfing in epiphytic bromeliads compensates for the limited pollination services provided by nectarivorous bats in a neotropical montane forest.

Abstract. Plants with specialized pollination systems frequently exhibit adaptations for self-pollination, and this contradictory situation has been explained in terms of the reproductive assurance function of selfing. In the neotropics, several plant lineages rely on specialized vertebrate pollinators for sexual reproduction, including the highly diverse Bromeliaceae family, which also displays a propensity for selfing. Thus far, the scarce evidence on the role of selfing in bromeliads and in other neotropical plant groups is inconclusive. To provide insights into the evolution and persistence of self-fertilization in the breeding systems of Bromeliaceae, we studied four sympatric epiphytic species from the genus Werauhia (Tillandsioideae) in Costa Rica. We documented their floral biology, pollination ecology and breeding systems. We estimated the contribution of selfing by comparing the reproductive success between emasculated flowers requiring pollinator visits and un-manipulated flowers capable of selfing and exposed to open pollination across two flowering seasons. The studied species displayed specialized pollination by nectar-feeding bats as well as a high selfing ability (auto-fertility index values > 0.53), which was attained by a delayed selfing mechanism. Fruit set from natural cross-pollination was low (<26% in both years) and suggested limited pollinator visitation. In line with this, we found a very low bat visitation to flowers using video-camera recording, from 0 to 0.24 visits per plant per night. On the contrary, the contribution of selfing was comparatively significant since 54-80% of the fruit set from un-manipulated flowers can be attributed to autonomous self-pollination. We concluded that inadequate cross-pollination services diminished the reproductive success of the studied Werauhia, which was compensated for by a delayed selfing mechanism. The low negative effects of inbreeding on seed set and germination likely reinforce the persistence of selfing in this bromeliad group. These results suggest that selfing in bat-pollinated bromeliads may have evolved as a response to pollinator limitation.

Florivory can facilitate rain-assisted autogamy in a deceptive tropical orchid.

Florivores and rainfall generally have negative impacts on plant fecundity. However, in some cases, they can mediate fruit set. Some plants face severe pollen-limited fecundity and any additional fruit set, even if from self-pollination, can be advantageous. This is the case in some tropical deceptive orchids, such as the threatened Cyrtopodium hatschbachii. Here we test the hypothesis that florivory of the anther cap would facilitate rain-assisted autogamy in this species. In the field, we followed flowers in which the anther cap was removed by the orthopteran Stenopola sp. and found cases where pollinia self-deposited after rainfall and in one case this resulted in swelling of the column typical of fruit development. This event comprised 33% of all fruit set in the population in 2019. We then experimentally varied anther cap removal and rainfall in a factorial design and found increased fruit set in the group with cap removal (simulated florivory) followed by rain. The water absorption by pollinia makes them heavier, causing the stipe to bend. The droplet of water on the stigma then shrinks and pulls the pollinia back onto the stigma, causing self-pollination. Seeds from self-pollination have considerable viability and may allow population persistence, given that bee-mediated cross-pollination is uncertain and even absent in some years. Our study provides a unique example of how two unrelated factors (i.e., florivory and rain) that are detrimental alone may together promote fruit set.

Mates Matter: Gametophyte Kinship Recognition and Inbreeding in the Giant Kelp, Macrocystis pyrifera (Laminariales, Phaeophyceae).

Inbreeding, the mating between genetically related individuals, often results in reduced survival and fecundity of offspring, relative to outcrossing. Yet, high inbreeding rates are commonly observed in seaweeds, suggesting compensatory reproductive traits may affect the costs and benefits of the mating system. We experimentally manipulated inbreeding levels in controlled crossing experiments, using gametophytes from 19 populations of Macrocystis pyrifera along its Eastern Pacific coastal distribution (EPC). The objective was to investigate the effects of male-female kinship on female fecundity and fertility, to estimate inbreeding depression in the F1 progeny, and to assess the variability of these effects among different regions and habitats of the EPC. Results revealed that the presence and kinship of males had a significant effect on fecundity and fertility of female gametophytes. Females left alone or in the presence of sibling males express the highest gametophyte size, number, and size of oogonia, suggesting they were able to sense the presence and the identity of their mates before gamete contact. The opposite trend was observed for the production of embryos per female gametes, indicating higher costs of selfing and parthenogenesis than outcrossing on fertility. However, the increased fecundity compensated for the reduced fertility, leading to a stable overall reproductive output. Inbreeding also affected morphological traits of juvenile sporophytes, but not their heatwave tolerance. The male-female kinship effect was stronger in high-latitude populations, suggesting that females from low-latitude marginal populations might have evolved to mate with any male gamete to guarantee reproductive success.

Fire frequency effects on cleistogamy expression and progeny performance in Cologania broussonetii.

Increased fire frequency usually erodes microenvironmental conditions, causing a drastic limitation of edaphic resources. Thus, the production of permanently closed-small flowers (cleistogamous, CL) should increase in sites with high fire frequency as this implies a less expensive reproductive assurance strategy. However, because open, insect-pollinated flowers (chasmogamous, CH) have the potential capacity to outcross via pollinators, CH progeny produced at any site should outperform selfed CL progeny. We evaluate the effect of fire frequency on the relative production of CL/CH flowers and fruits, and their seed set, along with several progeny performance parameters in Cologania broussonetii (Fabaceae), a resprouting herb with dimorphic cleistogamy native to the Chaco Serrano. Fire frequency increased cleistogamy expression, reaching extreme levels in high fire frequency sites. Seed set was similarly high for both CH and CL flowers in the unburned condition, while in burned sites the few developed CH flowers set more seeds than CL flowers. However, progeny performance was similar between CH and CL progeny at each and across all fire frequency conditions. Cleistogamy expression in C. broussonetii is maximized in abiotically degraded frequently burned habitats, although the selfed CL progeny is as successful as potentially outcrossed CH progeny. Fire frequency may decreased floral size and abundance, selecting for autogamous reproduction, which restricts not only the genetic potential of plant populations but also the resources offered to pollinators. At the community level, increased cleistogamy expression may potentially have negative implications for non-cleistogamous, more outcrossing species surviving in frequently burned environments.

Pollinator-independent orchid attracts biotic pollinators due the production of lipoidal substances.

Flowering plants often depend on the attraction of biotic pollinators for sexual reproduction. Consequently, the emergence and maintenance of selected floral attributes related to pollinator attraction and rewarding are driven by pollinator pressure. In this paper we explore the effect of pollinators, rainfall, temperature and air humidity on the reproduction of a Brazilian terrestrial orchid, Cranichis candida based on data of phenology, flower resources, olfactory and visual attraction cues, pollinators and breeding system. The flowers of C. candida are strongly protandrous and pollinated by workers of the social native bee Tetragonisca angustula. The bees collect labellar lipoidal substances (wax scales), which are transported to the nest. The lipoidal substance is composed of sterols, hydrocarbons and terpenes. The last presumably protects the bees and their nests against pathogens and other arthropods. C. candida sets fruits through biotic self- and cross-pollination, and spontaneously due the action of raindrops on flowers. Our results indicate that in C. candida, although rain-mediated spontaneous self-pollination happens, fructification mediated by biotic pollinations also occurs, which may result in fruit set by cross-pollination. A mixed pollination system must result in higher genetic variability when compared to species whose fruits are produced entirely by self-pollination. On the other hand, autogamy is a form of reproductive assurance, and has commonly evolved where pollination services are rare or absent.

Flower-level developmental plasticity to nutrient availability in Datura stramonium: implications for the mating system.

Background and Aims: Studies of phenotypic plasticity in plants have mainly focused on (1) the effect of environmental variation on whole-plant traits related to the number of modules rather than on (2) the phenotypic consequences of environmental variation in traits of individual modules. Since environmental and developmental factors can produce changes in traits related to the mating system, this study used the second approach to investigate whether within-individual variation in herkogamy-related traits is affected by the environment during plant development in two populations of Datura stramonium , an annual herb with a hypothesized persistent mixed mating system, and to determine which morphological traits may promote self-fertilization. Methods: Full-sib families of two Mexican populations of D. stramonium , with contrasting ecological histories, were grown under low, mid and high nutrient availability to investigate the effects of genetic, environmental and within-plant flower position on flower size, corolla, stamen and pistil lengths, and herkogamy. Key Results: Populations showed differences in familial variation, plasticity and familial differences in plasticity in most floral traits analysed. In one population (Ticumán), the effect of flower position on trait variation varied among families, whereas in the other (Pedregal) the effect of flower position interacted with the nutrient environment. Flower size varied with the position of flowers, but in the opposite direction between populations in low nutrients; a systematic within-plant trend of reduction in flower size, pistil length and herkogamy with flower position increased the probability of self-fertilization in the Pedregal population. Conclusions: Besides genetic variation in floral traits between and within populations, environmental variation affects phenotypic floral trait values at the whole-plant level, as well as among flower positions. The interaction between flower position and nutrient environment can affect the plant's mating system, and this differs between populations. Thus, reductions in herkogamy with flower positions may be expected in environments with either low pollinator abundance or low nutrients.

High selfing capability and low pollinator visitation in the hummingbird-pollinated epiphyte Pitcairnia heterophylla (Bromeliaceae) at a Costa Rican mountain forest / Alta capacidad de autofecundación y baja visitación de polinizadores en la epífita polinizada por colibríes Pitcairnia heterophylla (Bromeliaceae) en un bosque montano en Costa Rica

AbstractPitcairnioideae is the second most diverse subfamily of bromeliads (Bromeliaceae), a group exclusive to tropical regions of the New World. Pitcairnioid bromeliads have floral traits assumed to promote outcrossing through biotic pollination systems; however, the reproductive biology of most of the species of this group has not been documented. Pitcairnia heterophylla is an epiphytic (seldom saxicolous) bromeliad occurring from Southern Mexico, into the Northern Andes. We studied the pollination and breeding system of P. heterophylla in an epiphytic population at a mountain forest in Costa Rica from January to April 2013. We performed hand pollination experiments (agamospermy, autonomous self-pollination, hand self-pollination and hand cross-pollination) on 89 flowers from 23 individuals (3-6 flowers per individual) in 2013 flowering season. Nectar production was measured on 18 unvisited flowers of six individuals with a hand-held refractometer. Simultaneously, floral visitors were recorded on eight individuals with trail cameras for a total of 918 hours (115 ± 52 hours per individual, mean ± SE). Under natural conditions, seed set (540.4 ± 55.2) was similar to manually selfed flowers (516.3 ± 41.5) and autonomously selfed flowers (521.1 ± 29.0), but lower to manually outcrossed flowers (670.2 ± 31.3). The flowers of P. heterophylla are self-compatible, capable of autonomous pollination, and non-agamospermous. Intrafloral self-pollination is facilitated by adichogamy and lack of floral herkogamy. The scentless red flowers of P. heterophylla with tubular corollas and nectar production suggested ornithophilic pollination which was confirmed by video recording of 46 hummingbird visits. The most common floral visitor was the short-billed hummingbird Lampornis calolaemus which accounted for 78 % of the visits. However, the visitation rate during the flowering season was low (0.6 visits per day per plant). Selfing in P. heterophylla might be explained as a mechanism of reproductive assurance and to reduce interspecific pollen flow with taxonomically unrelated plants.

ResumenLa subfamilia Pitcairnioideae es la segunda más diversa de las bromelias (Bromeliaceae), un grupo exclusivo de la región Neotropical. Las bromelias pitcairnioideas poseen rasgos florales que se asume promueven el exo-cruzamiento a través de sistemas de polinización biótica; sin embargo, la biología reproductiva de la mayoría de especies de este grupo no se ha documentado. Pitcairnia heterophylla es una bromelia epífita (raramente saxícola) que se encuentra en el sur de México, América Central y el norte de los Andes. Se estudió la polinización y el sistema de apareamiento de una población P. heterophylla en un bosque montano en Costa Rica entre enero y abril 2013. Se realizaron cuatro experimentos de polinización (agamospermia, auto-polinización espontánea, auto-polinización manual, exo-polinización manual) en 89 flores de 23 individuos (3-6 flores por individuo). Se cuantificó la producción de néctar de 18 flores en seis individuos con un refractómetro. Simultáneamente, se registraron los visitantes florales de ocho individuos en el campo con ayuda de cámaras trampa por un total de 918 horas (115 ± 52 horas por individuo, promedio ± ES). Bajo condiciones naturales la producción de semillas fue similar (540.4 ± 55.2) a la producción de las flores auto-polinizadas manualmente (516.3 ± 41.5) y las flores auto-polinizadas espontáneamente (521.1 ± 29.0), pero fue menor a las flores exo-cruzadas (670.2 ± 31.3). Las flores de P. heterophylla son auto-compatibles, con alta capacidad de polinización espontánea y sin capacidad de agamospermia. La auto-polinización intra-floral es facilitada por la ausencia de dicogamia y hercogamia floral. Las flores rojas, sin aroma, con corola tubular y con producción de néctar sugirieron polinización por ornitofilia, lo que se confirmó con el registro de 46 visitas por colibríes (Apodiformes: Trochilidae). El visitante floral más común fue Lampornis calolaemus (78 % de las visitas). A pesar de esto, la tasa de visitas durante el periodo de estudio fue baja (0.6 visitas por día por planta). La capacidad de autofecundación de P. heterophylla puede ser explicada como un mecanismo de aseguramiento reproductivo, así como para reducir el flujo de polen inter-específico de plantas taxonómicamente no relacionadas.

Reproductive biology and pollination ecology of Triplaris gardneriana (Polygonaceae): a case of ambophily in the Brazilian Chaco.

Triplaris gardneriana (Polygonaceae) is a dioecious pioneer tree reported as insect-pollinated, despite possessing traits related to anemophily. Here, we analyse the possible roles of insects and wind on the pollination of this species to establish whether the species is ambophilous. We carried out observations of floral biology, as well as on the frequency and behaviour of pollinators visiting flowers in a population of T. gardneriana in the Chaco vegetation of Brazil. We conducted experimental pollinations to determine the maternal fertility of female plants and whether they were pollen-limited, and we also conducted aerobiological experiments to provide evidence of how environmental factors influence atmospheric pollen dispersal. The population comprised an area of approximately 152.000 m2 and was composed of 603 female and 426 male plants (sex ratio = 0.59:0.41). We observed 48 species of insects visiting flowers of T. gardneriana, of which the bees Scaptotrigona depilis and Apis mellifera scutellata were the most effective pollinators. We recorded pollen grains dispersed by wind on 74% of the glass slides placed on females, located at different distances (1-10 m) from male plants. Airborne pollen concentration was negatively correlated with relative humidity and positively correlated with temperature. Our observations and experimental results provide the first evidence that T. gardneriana is an ambophilous species, with pollen dispersal resulting from both animal and wind pollination. This mixed pollination strategy may be adaptive in T. gardneriana providing reproductive assurance during colonisation of sites with different biotic and abiotic conditions.

Evolutionary consequences of ecological factors: pollinator reliability predicts mating-system traits of a perennial plant.

The reproductive-assurance hypothesis predicts that mating-system traits will evolve towards increased autonomous self-pollination in plant populations experiencing unreliable pollinator service. We tested this long-standing hypothesis by assessing geographic covariation among pollinator reliability, outcrossing rates, heterozygosity and relevant floral traits across populations of Dalechampia scandens in Costa Rica. Mean outcrossing rates ranged from 0.16 to 0.49 across four populations, and covaried with the average rates of pollen arrival on stigmas, a measure of pollinator reliability. Across populations, genetically based differences in herkogamy (anther-stigma distance) were associated with variation in stigmatic pollen loads, outcrossing rates and heterozygosity. These observations are consistent with the hypothesis that, when pollinators are unreliable, floral traits promoting autonomous selfing evolve as a mechanism of reproductive assurance. Extensive covariation between floral traits and mating system among closely related populations further suggests that floral traits influencing mating systems track variation in adaptive optima generated by variation in pollinator reliability.

Variation in pollen limitation and floral parasitism across a mating system transition in a Pacific coastal dune plant: evolutionary causes or ecological consequences?

BACKGROUND AND AIMS: Evolutionary transitions from outcrossing to self-fertilization are thought to occur because selfing provides reproductive assurance when pollinators or mates are scarce, but they could also occur via selection to reduce floral vulnerability to herbivores. This study investigated geographic covariation between floral morphology, fruit set, pollen limitation and florivory across the geographic range of Camissoniopsis cheiranthifolia, a Pacific coastal dune endemic that varies strikingly in flower size and mating system. METHODS: Fruit set was quantified in 75 populations, and in 41 of these floral herbivory by larvae of a specialized moth (Mompha sp.) that consumes anthers in developing buds was also quantified. Experimental pollen supplementation was performed to quantify pollen limitation in three large-flowered, outcrossing and two small-flowered, selfing populations. These parameters were also compared between large- and small-flowered phenotypes within three mixed populations. KEY RESULTS: Fruit set was much lower in large-flowered populations, and also much lower among large- than small-flowered plants within populations. Pollen supplementation increased per flower seed production in large-flowered but not small-flowered populations, but fruit set was not pollen limited. Hence inadequate pollination cannot account for the low fruit set of large-flowered plants. Floral herbivory was much more frequent in large-flowered populations and correlated negatively with fruit set. However, florivores did not preferentially attack large-flowered plants in three large-flowered populations or in two of three mixed populations. CONCLUSIONS: Selfing alleviated pollen limitation of seeds per fruit, but florivory better explains the marked variation in fruit set. Although florivory was more frequent in large-flowered populations, large-flowered individuals were not generally more vulnerable within populations. Rather than a causative selective factor, reduced florivory in small-flowered, selfing populations is probably an ecological consequence of mating system differentiation, with potentially significant effects on population demography and biotic interactions.

Ecologia da interação entre as pequenas flores de quilha de Stylosanthes viscosa Sw. (Faboideae) e as grandes abelhas Xylocopa (Neoxylocopa) cearensis Ducke, 1910 (Apoidea, Hymenoptera), em duna tropical / Ecological interaction between the tiny keel flowers of Stylosanthes viscosa Sw. (Faboideae) and the large bee Xylocopa (Neoxylocopa) cearensis Ducke, 1910 (Apoidea, Hymenoptera), in tropical sand dune

Stylosanthes viscosa is a common 'herb' in the tropical sand dunes of the Brazilian coast with clumped distribution in urban sand dunes of Salvador, BA. Its tiny keel flower with a very small amount of nectar hidden inside the corolla tube, a single open flower/inflorescence/day and the low floral density altogether should not be attractive to large bees. This should be true mainly upon the high-demanded energy for thermoregulation within the tropical sand dunes. Contrary to this expectation, most of the bee species sampled on S. viscosa flowers were medium to large sized bees and harvested nectar in legitimate visits by activating the keel's pollination mechanism of flowers. However, the keel was always inactivated after being visited by the enormous and frequent (78 percent) Xylocopa bees. All the flowers visited by these bees (including those with inactivated keels), and those experimentally protected from bee visiting produced fruits (100 percent). The specialized embolus mechanism of the anthers assures self-pollination. Because of this autonomous self-pollination and self-compatibility it was not possible to discriminate the relative importance of both facilitated self-pollination and cross-pollination by bees. Nevertheless, the large Xylocopa bees didn't limit self-pollination. Xylocopa cearensis was the most abundant bee (63 percent) and it visited several flowers (13.0 ± 4.27) in each flight route in the same clump, in spite of the low floral density therein (5 to 20 flowers.m-2). Flight distances between successive flowers varied between one and two meters (49 percent), and so they were higher than would be possible (15 to 70 cm) given the floral density in the clumps. The flight routes were directional, and most of shifting direction between successive flowers was below 90° with high frequency of small angles (<30°). This general pattern is explained by foraging optimization with adjustments to critical resource level thresholds. Paradoxically, floral biology and flowering pattern of S. viscosa should discourage the largest dune dweller bees, but this strategy works just temporarily. In this melittophilous species, the autonomous self-pollination (assured by the embolus) can be better explained by the automatic selection hypothesis rather than by reproductive assurance hypothesis, considering the availability of large bee pollinators in the tropical dunes.

Stylosanthes viscosa é uma herbácea comum nas dunas tropicais da costa brasileira, com distribuição agregada nas dunas urbanas de Salvador, BA. Suas flores de quilha diminutas, com uma pequena quantidade de néctar escondido no tubo da corola, a abertura de uma única flor/inflorescência/dia e a baixa densidade floral, juntos, não deveriam ser atrativos para as abelhas de grande porte. Isso deve ser verdadeiro, principalmente, sob a alta demanda energética para termorregulação em dunas tropicais. Ao contrário do esperado, a grande maioria das espécies de abelhas amostradas nas flores de S. viscosa tem tamanho médio a grande e coletou néctar em visitas legítimas, acionando o mecanismo de polinização da quilha. Entretanto, a quilha sempre ficava inutilizada após as visitas das grandes e frequentes (78 por cento) abelhas Xylocopa. Todas as flores visitadas por essas abelhas (inclusive aquelas com a quilha inutilizada) e as protegidas experimentalmente das visitas das abelhas produziram frutos regularmente (100 por cento). O mecanismo de êmbolo das anteras assegura a autopolinização. Como há autopolinização autônoma e o sistema é autocompatível ainda não foi possível discriminar a importância relativa da autopolinização facilitada e da polinização cruzada por abelhas. Entretanto, as grandes abelhas Xylocopa não limitam a autopolinização. Xylocopa cearensis foi a abelha mais abundante (63 por cento) e visitou várias flores (13,0 ± 4,27) em cada rota de vôo, numa mesma mancha, apesar da baixa densidade floral (5 a 20 flores.m-2). A distância de vôo entre flores sucessivas variou entre um e dois metros (49 por cento), acima do que seria possível (15 a 70 cm) dada a densidade floral nas manchas. As rotas de vôo foram direcionais, isto é, a maioria dos ângulos de mudança de direção entre flores sucessivas foram abaixo de 90°, com alta frequência de ângulos pequenos (<30°). Este padrão geral é explicado com a otimização de forrageio, com ajuste a limiares críticos de oferta de recursos. Paradoxalmente, a biologia floral e o padrão de florescimento de S. viscosa deveriam desestimular a visita dessas grandes abelhas, mas esta estratégia funciona apenas temporariamente. Nesta espécie melitófila, a autopolinização autônoma (assegurada pelo êmbolo) pode ser melhor explicada pela hipótese de seleção automática do que pela hipótese de segurança, considerando a disponibilidade de abelhas grandes como polinizadores nas dunas tropicais.