PARASITES AND THE EVOLUTION OF SELF-FERTILIZATION

Abstract.— Assuming all else is equal, an allele for selfing should spread when rare in an outcrossing population and rapidly reach fixation. Such an allele will not spread, however, if self‐fertilization results in inbreeding depression so severe that the fitness of selfed offspring is less that half that of outcrossed offspring. Here we consider an ecological force that may also counter the spread of a selfing allele: coevolution with parasites. Computer simulations were conducted for four different genetic models governing the details of infection. Within each of these models, we varied both the level of selfing in the parasite and the level of male‐gamete discounting in the host (i.e., the reduction in outcrossing fitness through male function due to the selfing allele). We then sought the equilibrium level of host selfing under the different conditions. The results show that, over a wide range of conditions, parasites can select for host reproductive strategies in which both selfed and outcrossed progeny are produced (mixed mating). In addition, mixed mating, where it exits, tends to be biased toward selfing.     

Experimental and genetic analyses reveal that inbreeding depression declines with increased self‐fertilization among populations of a coastal dune plant

Theory predicts that inbreeding depression (ID) should decline via purging in self‐fertilizing populations. Yet, intraspecific comparisons between selfing and outcrossing populations are few and provide only mixed support for this key evolutionary process. We estimated ID for large‐flowered (LF), predominantly outcrossing vs. small‐flowered (SF), predominantly selfing populations of the dune endemic Camissoniopsis cheiranthifolia by comparing selfed and crossed progeny in glasshouse environments differing in soil moisture, and by comparing allozyme‐based estimates of the proportion of seeds selfed and inbreeding coefficient of mature plants. Based on lifetime measures of dry mass and flower production, ID was stronger in nine LF populations [mean δ = 1?(fitness of selfed seed/fitness of outcrossed seed) = 0.39] than 16 SF populations (mean δ = 0.03). However, predispersal ID during seed maturation was not stronger for LF populations, and ID was not more pronounced under simulated drought, a pervasive stress in sand dune habitat. Genetic estimates of δ were also higher for four LF (δ = 1.23) than five SF (δ = 0.66) populations; however, broad confidence intervals around these estimates overlapped. These results are consistent with purging, but selective interference among loci may be required to maintain strong ID in partially selfing LF populations, and trade‐offs between selfed and outcrossed fitness are likely required to maintain outcrossing in SF populations.     

OUTCROSSING RATE AND INBREEDING DEPRESSION IN TWO ANNUAL MONOECIOUS HERBS,BEGONIA HIRSUTA AND B. SEMIOVATA

Most models of mating-system evolution predict inbreeding depression to be low in inbred populations due to the purging of deleterious recessive alleles. This paper presents estimates of outcrossing rates and inbreeding depression for two highly selfing, monoecious annuals Begonia hirsuta and B. semiovata. Outcrossing rates were estimated using isozyme polymorphisms, and the magnitude of inbreeding depression was quantified by growing progeny in the greenhouse produced through controlled selfing and outcrossing. The estimated single-locus outcrossing rate was 0.03 ± 0.01 (SE) for B. hirsuta and 0.05 ± 0.02 for B. semiovata. In both species, the seed production of selfed flowers was on average 12% lower than that of outcrossed flowers (B. hirsuta P = 0.07, B. semiovata P < 0.05, mixed model ANOVAs). There was no significant effect of crosstype on germination rate or survival, but selfed offspring had a lower dry mass than outcrossed offspring 18 weeks after planting in both species (on average 18% lower in B. hirsuta and 31% lower in B. semiovata). Plants that were the products of selfing began flowering later than plants produced through outcrossing in B. semiovata, but not in B. hirsuta. The effects of crosstype on seed production (B. semiovata), days to first flower and offspring dry mass (both species) varied among maternal parents, as indicated by significant crosstype x maternal parent interactions for these characters. Both species showed significant inbreeding depression for total fitness (estimated as the product of seed production, germination rate, survival and dry mass at 18 weeks). In B. hirsuta, the average total inbreeding depression was 22% (range -57%-98%; N = 23 maternal parents), and in B. semiovata, it was 42% (-11%-84%; N = 21). This study demonstrates that highly selfing populations can harbor substantial inbreeding depression. Our findings are consistent with the hypothesis that a high mutation rate to mildly deleterious alleles contributes to the maintenance of inbreeding depression in selfing populations.     

Evolution of the Selfing Rate and Resource Allocation Models

Abstract First, evolutionary theories of selfing of terrestrial plants are reviewed briefly. The evolution of the selfing rate is controlled mainly by (1) the benefit of enhanced genetic relatedness to seeds and (2) the cost of lowered fitness of selfed offspring (inbreeding depression), being modified by (3) fertility assurance under pollen limitation, (4) reduced performance as pollen donor, (5) reduced expenditure to male function, and (6) lowered genetic recombination. Models of the joint evolution of selfing and inbreeding depression predict either strong outcrossing or predominant selfing. Although wind-pollinated plants fit the prediction, some animal-pollinated species have intermediate selfing rates, refuting the theory.
Second, three resource allocation models are analyzed, in which an individual plant optimally allocates limited resources to outcrossed seeds, selfed seeds, and to energy reserves for the next year. The first model explains how the number of outcrossed and selfed offspring change with plant size when they differ in dispersal distance. The second model predicts that, in a disturbed habitat, the plant is likely to be annual and to produce both selfed and outcrossed seeds; in contrast, in a stable habitat, the plant tends to be perennial and to abort selfed seeds selectively. Hand pollination may increase seed production for perennials but not for annuals. The third model explains the observed difference between animal and wind pollinated plants in the out-crossing rate pattern by the difference in the way pollen acquisition increases with investment.     

The mixed mating system of Impatiens capensis and infection by a foliar rust pathogen: patterns of resistance and fitness consequences

Outcrossing by hosts may offer protection from natural enemies adapted to parental genotypes by creating diverse progeny that differ from their parents through genetic recombination. However, past experimental work addressing the relationship between mating system and disease in offspring has given conflicting results, suggesting that outcrossing might also cause the dissolution of resistant genotypes. To determine if selfed progeny are more susceptible to disease caused by the heteroecious rust, Puccinia recondita, or if selfing preserves existing resistant genotypes, we used a factorial design to compare levels of infection of selfed and outcrossed progeny of Impatiens capensis, a woodland annual with a mixed mating system. We compared the level of host infection when exposed to three pathogen sources in the field: the sympatric rust population, and two allopatric rust populations. Outcrossed progeny exposed to sympatric rust had higher infection scores than selfed progeny exposed to the same rust, suggesting that outcrossing breaks up resistant genotypes. In addition, there was a trend for the rust to be more infective on sympatric rather than allopatric hosts. We also examined whether rust infection differentially alters the fitness of selfed and outcrossed progeny. Outcrossed plants that escaped infection had higher fitness, as measured by fruit production, than selfed plants, but there was no difference in fitness between infected selfed and infected outcrossed plants. Thus, outcrossing was advantageous in the absence of disease, but there was no fitness difference between selfed and outcrossed progeny in the presence of disease. In sum, our results indicate that interactions with pathogens can eliminate or reverse the advantage of outcrossing.     

Sibling competition, size variation and frequency-dependent outcrossing advantage in Plantago coronopus

Many plants display limited seed dispersal, thereby creating an opportunity for sibling competition, i.e. fitness-determined interactions between related individuals. Here I investigated the consequences of intra-specific competition, by varying density and genetic composition of neighbors, on the performance of seedlings derived by selfing or outcrossing of the partially self-fertilizing plant Plantago coronopus (L.). Seedlings from eight plants, randomly selected from an area of about 50 m² in a natural population, were used in (i) a density series with either one, four or eight siblings of each cross type per pot and (ii) a replacement series with eight plants per pot where selfed and outcrossed siblings were grown intermixed in varying frequencies. Density had a pronounced effect on plant performance. But, except for singly grown individuals, no differences were detected between selfed and outcrossed progenies in vegetative and reproductive biomass. When grown intermixed, selfed offspring were always inferior to their outcrossed relatives. The magnitude of reduction in performance was dependent on the number of outcrossed relatives a selfed seedling had to compete with, giving rise to a frequency-dependent fitness advantage to outcrossed seedlings. The major result of this study is (i) that the relative fitness of inbred progeny is strongly affected by the type of competitors (inbred or outbred) and (ii) that inbreeding depression varies according to the density and frequency of outbred plants and could be considered as a density- and frequency-dependent phenomenon. It is argued that sibling competition, due to the small genetic neighborhood of P. coronopus, might be an important selective force in natural populations of this species.     

Different levels of inbreeding depression between outcrossing and selfing Serapias species

We quantified inbreeding depression for fruit production, embryo vitality and seed germination in three deceptive orchids, Serapias vomeracea, S. cordigera and S. parviflora, which do not provide any reward to their pollinators, and are predicted to experience high outcrossing. Of the three species examined only S. parviflora was autonomously selfing. Both S. vomeracea and S. cordigera showed highly significant differences in fitness between selfed and outcrossed progenies, resulting in high levels of inbreeding depression, which increased in magnitude from seed set to seed germination. Inbreeding depression may promote outcrossing in Serapias by acting as a post-pollination barrier to selfing. Cumulative inbreeding depression across three stages in S. parviflora was lower that in both outcrossing species. The large difference in germination between selfed and outcrossed seeds is an important issue in conservation biology.     

Individualized mating system estimation using genomic data

The estimation of outcrossing rates in hermaphroditic species has been a major focus in the evolutionary study of reproductive strategies, and is also essential for plant breeding and conservation. Surprisingly, genomics has thus far minimally influenced outcrossing rate studies. In this article, we generalize a Bayesian inference method (BORICE) to accommodate genomic data from multiple subpopulations of a species. As an empirical demonstration, BORICE is applied to 115 maternal families of Mimulus guttatus. The analysis shows that low‐level whole genome sequencing of parents and offspring is sufficient for individualized mating system estimation: 208 offspring (88.5%) were definitively called as outcrossed, 23 (9.8%) as selfed. After mating system parameters are established (each offspring as outcrossed or selfed and the inbreeding level of maternal plants), BORICE outputs posterior genotype probabilities for each SNP genomewide. Individual SNP calls are often burdened with considerable uncertainty and distilling information from closely linked sites (within genomic windows) can be a useful strategy. For the Mimulus data, principal components based on window statistics were sufficient to diagnose inversion polymorphisms and estimate their effects on spatial structure, phenotypic and fitness measures. More generally, mating system estimation with BORICE can set the stage for population and quantitative genomic analyses, particularly researchers collect phenotypic or fitness data from maternal individuals.     

THE RELATIVE COSTS OF SELF- AND CROSS-FERTILIZED SEEDS IN IMPATIENS CAPENSIS (BALSAMINACEAE)

By using a generally applicable technique that involves monitoring the development and survivorship of flowers and seed capsules, I estimated the material and energetic costs of producing self- and cross-fertilized seeds in Impatiens capensis. All flowers and fruits on six plants were censused intensively for the two-month period of reproduction. Cleistogamous (selfing) flowers ripened seed in about 24 days, compared to about 36 days for the chasmogamous (outcrossing) flowers. In terms of dry weight, selfed seeds cost about two-thirds as much as outcrossed seeds: 12.4 versus 18.4 mg dry weight per seed. When adjusted to the currency of calories, and including an independent estimate of pollen and nectar production in outcrossing flowers, I estimate the costs to be about 65 and 135 calories per selfed or outcrossed seed. Sources of error include the accuracy of the estimates of flower and fruit weight, and possible differences among the developmental stages in respiratory costs. The cost discrepancy implies that outcrossed seeds should possess a countervailing fitness advantage large enough to offset their greater energetic cost.     

Experimental analysis of mating patterns in a clonal plant reveals contrasting modes of self‐pollination

Hermaphrodite plants commonly practice self‐fertilization (selfing), but the mechanisms responsible vary depending on the mode of self‐pollination, pollinator behavior, and degree of clonality. Whether selfing occurs within (autogamy) or between flowers (geitonogamy) is of evolutionary significance because their fitness consequences differ. We used floral manipulations and genetic markers to determine the relative contribution of autogamy and within‐ versus between‐ramet geitonogamy to the selfing rate of the bumblebee‐pollinated, clonal herb Aconitum kusnezoffii. Data on flowering phenology and bumblebee foraging were also collected to determine opportunities for different modes of self‐pollination. Autogamy accounted for only 12% of the selfing rate with the remainder resulting from geitonogamy. Whole‐ramet emasculation of clones with multiple ramets reduced selfing by 78%, indicating that within‐ramet geitonogamy contributed significantly (68%) to total selfing. Selfing of single‐ramet plants was 44% less than multiple‐ramet plants, indicating that the contribution of between‐ramet geitonogamy was substantially less (20%) than within‐ramet geitonogamy, probably because of bumblebee foraging behavior. Our results demonstrate for the first time in a clonal plant that within‐ramet geitonogamy is substantially greater than between‐ramet geitonogamy and highlight the importance of considering the influence of clonal architecture and pollinator foraging on modes of self‐pollination.     

INBREEDING DEPRESSION IN PARTIALLY SELF-FERTILIZING DECODON VERTICILLATUS (LYTHRACEAE): POPULATION-GENETIC AND EXPERIMENTAL ANALYSES

Inbreeding depression is a major selective force favoring outcrossing in flowering plants. Some self-fertilization, however, should weaken the harmful effects of inbreeding by exposing genetic load to selection. This study examines the maintenance of inbreeding depression in partially self-fertilizing populations of the long-lived, herbaceous wetland plant, Decodon verticillatus (L.) Ell. (Lythraceae). Estimates from ten populations indicate that 30% of offspring are produced through self-fertilization. Population-genetic estimates of inbreeding depression (δ = 1 – relative mean fitness of selfed progeny) based on changes in the inbreeding coefficient for the same ten populations were uniformly high, ranging from 0.49 to 1.79 and averaging 1.11 ± 0.29 SE. Although confidence intervals of individual population estimates were large, estimates were significantly greater than 0 in six populations and greater than 0.5 in four. Inbreeding depression was also estimated by comparing growth, survival, and flowering of experimentally selfed and outcrossed offspring from two of these populations in a 1-yr glasshouse experiment involving three density regimes; after which offspring were transplanted into garden arrays and two field sites and monitored for two consecutive growing seasons. Overall for survival averaged 0.27 ± 0.01 in the glasshouse, 0.33 ± 0.04 in the garden, and 0.46 ± 0.04 in the field. The glasshouse experiment also revealed strong inbreeding depression for growth variables, especially above-soil dry weight ( = 0.42 ± 0.03). The fitness consequences of inbreeding depression for these growth variables approximately doubles if survival to maturity is determined by severe truncation selection. Despite substantial selfing, inbreeding depression appears to be a major selective force favoring the maintenance of outcrossing in D. verticillatus.     

No inbreeding depression in an outcrossing alpine species: The breeding system of Campanula thyrsoides

Plants that live in fragmented landscapes, where populations are isolated from each other and in which long-distance dispersal is essential for colonization of empty sites, reproduction should be favoured by self-compatibility (Baker's law). Nevertheless, outcrossing mechanisms, such as self-incompatibility and dichogamy, are common in many species and are often maintained by inbreeding depression in the fitness of selfed progeny. Here, we studied the breeding system and the consequences of selfing and sister mating in Campanula thyrsoides, a short-lived perennial monocarp, which is found in the naturally fragmented landscape of the Alps. An experiment with controlled pollinations was set up in the common garden with plants grown from seeds originating from 14 seed families, collected in the siliceous Central Alps, where this plant is found on isolated carbonate bearing outcrops.Our results indicate that C. thyrsoides has a strong self-incompatibility system (SI) with no or low seed set in selfed flowers compared to outcrossed and sister-crossed flowers. Moreover, the SI system in C. thyrsoides did not break down with flower age as in some other Campanula species. Surprisingly, there was no significant difference in seed set, seed weight, germination percentage, seedling survival and size between outcrossed and sister-crossed offspring, which indicates no inbreeding depression.We suggest that the absence of inbreeding depression in this outcrossing species might be a result of frequent bottlenecks during colonization of the isolated habitats in the alpine landscape.     

Effects of reproductive compensation, gamete discounting and reproductive assurance on mating-system diversity in hermaphrodites

Hermaphroditism allows considerable scope for contributing genes to subsequent generations through various mixtures of selfed and outcrossed offspring. The fitness consequences of different family compositions determine the evolutionarily stable mating strategy and depend on the interplay of genetic features, the nature of mating, and factors that govern offspring development. This theoretical article considers the relative contributions of these influences and their interacting effects on mating-system evolution, given a fixed genetic load within a population. Strong inbreeding depression after offspring gain independence selects for exclusive outcrossing, regardless of the intensity of predispersal inbreeding depression, unless insufficient mating limits offspring production. The extent to which selfing evolves under weak postdispersal inbreeding depression depends on predispersal inbreeding depression and the opportunity for resource limitation of offspring production. Mixed selfing and outcrossing is an evolutionarily stable strategy (ESS) if selfed zygotes survive poorly, but selfed offspring survive well, and maternal individuals produce enough "extra" eggs that deaths of unviable outcrossed embryos do not impact offspring production (reproductive compensation). Mixed mating can also be an ESS, despite weak lifetime inbreeding depression, if self-mating reduces the number of male gametes available for outcrossing (male-gamete discounting). Reproductive compensation and male-gamete discounting act largely independently on mating-system evolution. ESS mating systems always involve either complete fertilization or fertilization of enough eggs to induce resource competition among embryos, so although reproductive assurance is adaptive with insufficient mating, it is never an ESS. Our results illustrate the theoretical importance of different constraints on offspring production (availability of male gametes, egg production, and maternal resources) for both the course and outcome of mating-system evolution, whereas unequal competition between selfed and outcrossed embryos has limited effect. These results also underscore the significance of heterogeneity in the nature and intensity of inbreeding depression during the life cycle for the evolution of hermaphrodite mating systems.     

The inbreeding depression cost of selfing: Importance of flower size and population size in Collinsia parviflora (Veronicaceae)

Inbreeding depression should evolve with selfing rate when frequent inbreeding results in exposure of and selection against deleterious alleles. The selfing rate may be modified by plant traits such as flower size, or by population characteristics such as census size that can affect the probability of biparental inbreeding. Here we quantify inbreeding depression (δ) among different population sizes of Collinsia parviflora, a wildflower with interpopulation variation in flower size, by comparing fitness components and multiplicative fitness of experimentally produced selfed and outcrossed offspring. Selfed offspring had reduced multiplicative fitness compared to outcrossed offspring, but inbreeding depression was low in all combinations of population size and flower size (δ ≤ 0.05) except in large populations of large-flowered plants (δ = 0.45). The decrement to multiplicative fitness with inbreeding was not affected by population size nested within flower size, but differed between small- and large-flowered plants: small-flowered populations had lower overall inbreeding depression (δ = 0.04) compared to large-flowered populations (δ = 0.25). The difference in load with flower size suggests that either selection has removed deleterious recessive alleles or these alleles have become fixed in small-flowered, potentially more selfing populations, but that purging has not occurred to the same extent in presumably outcrossing large-flowered populations.     

Nonrandom mating in Clarkia gracilis (Onagraceae): a case of cryptic self-incompatibility

In plants capable of both self-fertilization and outcrossing, the selfing rate depends on the proportion of self pollen in pollen loads and on the relative postpollination success of self pollen in siring offspring. While the composition of pollen loads is subject to unpredictable variation, paternity success of self vs. outcross pollen following pollen deposition may be controlled by maternal plants. This study examined postpollination paternity success in Clarkia gracilis ssp. sonomensis, in which deposition of self pollen is common. Pure loads of self and outcross pollen produced similar numbers of mature seeds, but equal mixtures of self and outcross pollen yielded more than three times as many outcrossed offspring as selfed offspring. The finding that the paternity success of self pollen depends on whether it is in competition with outcross pollen helps to explain an earlier finding that the selfing rate in experimental populations was highest when pollinator activity was lowest. Cryptic self-incompatibility allows paternity by self pollen when outcross pollen is unavailable.     

The impact of extensive clonal growth on fine-scale mating patterns: a full paternity analysis of a lily-of-the-valley population (Convallaria majalis)

Background and Aims

The combination of clonality and a mating system promoting outcrossing is considered advantageous because outcrossing avoids the fitness costs of selfing within clones (geitonogamy) while clonality assures local persistence and increases floral display. The spatial spread of genetically identical plants (ramets) may, however, also decrease paternal diversity (the number of sires fertilizing a given dam) and fertility, particularly towards the centre of large clumped clones. This study aimed to quantify the impact of extensive clonal growth on fine-scale paternity patterns in a population of the allogamous Convallaria majalis.

Methods

A full analysis of paternity was performed by genotyping all flowering individuals and all viable seeds produced during a single season using AFLP. Mating patterns were examined and the spatial position of ramets was related to the extent of multiple paternity, fruiting success and seed production.

Key Results

The overall outcrossing rate was high (91 %) and pollen flow into the population was considerable (27 %). Despite extensive clonal growth, multiple paternity was relatively common (the fraction of siblings sharing the same father was 0·53 within ramets). The diversity of offspring collected from reproductive ramets surrounded by genetically identical inflorescences was as high as among offspring collected from ramets surrounded by distinct genets. There was no significant relationship between the similarity of the pollen load received by two ramets and the distance between them. Neither the distance of ramets with respect to distinct genets nor the distance to the genet centre significantly affected fruiting success or seed production.

Conclusions

Random mating and considerable pollen inflow most probably implied that pollen dispersal distances were sufficiently high to mitigate local mate scarcity despite extensive clonal spread. The data provide no evidence for the intrusion of clonal growth on fine-scale plant mating patterns.     

Influence of clonal growth on selfing rate in Vaccinium myrtillus L

Clonal growth, which allows the multiplication of flowering shoots of the same genet, can lead to a large floral display and may thus increase the rate of selfing through geitonogamy as a consequence of an increase in the number of successively visited flowers. The aim of the present research was to analyse the combined effect of the diversity and intermingling of clones on the rate of selfing in Vaccinium myrtillus. Four mother plants were selected within patches characterised by contrasting clonal structure (low versus high number and intermingling of clones). The selfing rate was significantly lower for plants situated within patches characterised by a high number of intermingled clones (3%) than for plants situated in patches with a low number of clones (50%). Therefore, for this species suffering from inbreeding depression, an increase in the number or the intermingling of the clones could reduce the rate and the cost of geitonogamy and allow a large floral display to attract pollinators. We also found that the main pollinators, bumblebee queens, presented a foraging behaviour favouring geitonogamy, as their successive visits to flowers were quite short (89% of flights were 40 cm or less).     

FITNESS CONSEQUENCES OF OUTCROSSING IN IMPATIENS CAPENSIS: TESTS OF THE FREQUENCY-DEPENDENT AND SIB-COMPETITION MODELS

Using field and greenhouse experiments, we tested two hypotheses that could account for the maintenance of outcrossing in Impatiens capensis. Seedlings derived from cleistogamous (CL) and chasmogamous (CH) flowers were grown under competitive conditions while flanked by neighbors that were either related or unrelated. In both experiments, CH progeny sometimes expressed more phenotypic variability than CL progeny. In the greenhouse experiment, CH progeny attained the same weight as CL progeny, and the relatedness of neighboring plants did not affect the growth of either type. In the field experiment, CL and CH progeny performed similarly when grown with related competitors. However, CH progeny were somewhat larger when planted with nonsibs, while CL progeny were somewhat smaller under those conditions. Thus, there is no evidence that either frequency-dependent selection or the avoidance of competition among siblings favors the maintenance of outcrossing in this species. We also modeled the relative variability of selfed and outcrossed progeny under several reproductive systems. When females mate with one male (progeny are full sibs), selfed progeny are often more variable than outcrossed progeny. When females engage in both selfing and outcrossing, variation among progeny is frequently maximized at an intermediate selfing rate. The sib-competition mechanism, under a range of genetic models, is not apt to promote outcrossing, since selfed progeny are commonly more variable than outcrossed progeny.     

BREEDING SYSTEMS OF CENTRAL AMERICAN APHELANDRA (ACANTHACEAE)

Controlled pollinations, followed by germination studies of seeds resulting from self- and cross-pollination, were carried out using plants from 12 populations of nine species of Aphelandra (Acanthaceae), a genus of neotropical shrubs and herbs. These results are combined with data from field studies of flower morphology, phenology, and pollinator relationships to estimate breeding system of each species. All species have floral morphological traits that prevent autogamy. Plants from five populations of five species experience low levels of geitonogamous pollen transfer; they produce few flowers daily and are pollinated by traplining hummingbirds. Excepting A. storkii, these plants are fully self-compatible (SC), and seeds from selfing are as viable as crossed seeds. Aphelandra storkii is partially self-incompatible (SI) and produces seeds from selfing that tend to germinate less successfully than crossed seeds. Plants from the remaining populations are profusely flowering shrubs, and even those pollinated by traplining hummingbirds should experience higher levels of geitonogamy. Aphelandra deppeana is pollinated by territorial hummingbirds, which should further increase the incidence of geitonogamy. All shrub species are partially SI, and two species (A. leonardii and A. sinclairiana) show significant reduced germination of selfed vs. crossed seeds. The breeding system of these species is thus modified by postpollination factors that favor the formation and maturation of outcrossed seeds. It is suggested that Aphelandra species, like other herbs and shrubs of tropical forest understory, possess a combination of breeding system traits that promote outcrossing but do not exclude geitonogamy.     

Effects of cleistogamy and pollen source on seed production and offspring performance in three endangered violets

In rare plants that often occur in small or isolated populations the probability of selfing between close relatives is increased as a consequence of demographic stochasticity. The mode of pollination (selfing, outcrossing) may have considerable effects on seed traits and offspring performance and hence potential viability. Since current efforts aiming at the restoration of floodplain grasslands through the transfer of plant material from species-rich source stands may lead to the establishment of initially small populations consisting of founders from different populations, the present paper experimentally investigated the effects of pollen source and floral types (i.e. chasmogamous (CH) and cleistogamous (CL) flowers) on seed traits and offspring performance in three highly endangered violet species (Viola elatior, V. pumila, V. stagnina) of these grasslands. We estimated inbreeding depression and tested the performance of selfed and outcrossed offspring in two microbial environments, i.e. in soil inoculated with (i) non-sterile substrate from the same species (‘home’-conditions) and (ii) sterilised substrate.Plants produced more CL capsules than CH flowers. Pollinator exclusion had only small effects on CH seed production. CL seeds had a significantly lower mass per seed than CH seeds. This may be related to constraints in allocation or environmental conditions. Seedling growth was reduced in plants grown under ‘home’-conditions as compared to control soils. Under ‘home’-conditions, relative fitness of selfed seedlings of V. stagnina was significantly higher than that of crossed progeny. Our results suggest that high genetic differentiation among populations as a consequence of isolation may result in outbreeding depression, e.g., through biochemical or physiological incompatibilities between genes or the breaking of coadapted gene complexes. In V. stagnina, offspring fitness differed considerably between environments, but in general we found no indications for inbreeding depression in these rare species.