Pollen limitation and inbreeding depression in an 'old rare' bumblebee-pollinated grassland herb

Habitat fragmentation and reduction of population size have been found to negatively affect plant reproduction in 'new rare' species that were formerly common. This has been attributed primarily to effects of increased inbreeding but also to pollen limitation. In contrast, little is known about the reproduction of 'old rare' species that are naturally restricted to small and isolated habitats and thus may have developed strategies to cope with long-term isolation and small population size. Here we study the effects of pollen source and quantity on reproduction of the 'old rare' bumblebee-pollinated herb, Astragalus exscapus. In two populations of this species, we tested for pollen autodeposition, inbreeding depression and outbreeding depression. Caged plants were left unpollinated or were pollinated with pollen from the same plant, from the same population or from a distant population (50 km). Additionally, we tested for pollen limitation by pollen supplementation in four populations of different size and density. In the absence of pollinators, plants did not produce seed whereas self-pollinated plants did. This indicates a self-compatible breeding system dependent on insect pollination. Both self-pollination and, in one of the two populations, cross-pollination with pollen from plants from the distant population resulted in a lower number of seeds per flower than cross-pollination with pollen from plants from the resident population, indicating inbreeding and outbreeding depression. Pollen addition enhanced fruit set and number of seeds per flower in three of the four populations, indicating pollen limitation. The degree of pollen limitation was lowest in the smallest but densest population. Our results suggest that, similar to 'new rare' plant species, also 'old rare' species may be at risk of inbreeding depression and pollen limitation.     

Mating structure and inbreeding and outbreeding depression in the rare plant Gentianella germanica (Gentianaceae)

Isolation and small size of populations as a result of habitat destruction and fragmentation may negatively affect plant fitness through pollinator limitation and increased levels of inbreeding. To increase genetic variation in small populations of rare plants artificial gene flow has been suggested as a management tool. We investigated whether pollinator limitation and inbreeding depression could reduce fitness in Gentianella germanica, an endangered biennial of increasingly fragmented calcareous grasslands in Central Europe. We experimentally excluded pollinators and generated progenies by hand-pollinating flowers with pollen from different distances. G. germanica was highly selfing. Pollinator exclusion strongly reduced seed set, indicating that pollinator limitation could potentially reduce plant fitness. Germination rate as well as number of leaves and rosette size of progeny from 10-m crosses was higher than that of progeny from open pollinations, self-, 1-m, and interpopulation crosses. After 6 mo of growth differences in the number of surviving plants persisted, whereas differences in plant size did not. The results suggest that inbreeding depression may reduce plant performance in G. germanica. Outbreeding depression in the performance of progeny from interpopulation crosses indicates that caution is necessary in using artificial interpopulation gene flow as a management tool.     

The relative importance of reproductive assurance and automatic selection as hypotheses for the evolution of self-fertilization

Background

The field of plant mating-system evolution has long been interested in understanding why selfing evolves from outcrossing. Many possible mechanisms drive this evolutionary trend, but most research has focused upon the transmission advantage of selfing and its ability to provide reproductive assurance when cross-pollination is uncertain. We discuss the shared conceptual framework of these ideas and their empirical support that is emerging from tests of their predictions over the last 25 years.

Scope

These two hypotheses are derived from the same strategic framework. The transmission advantage hypothesis involves purely gene-level selection, with reproductive assurance involving an added component of individual-level selection. Support for both of these ideas has been garnered from population-genetic tests of their predictions. Studies in natural populations often show that selfing increases seed production, but it is not clear if this benefit is sufficient to favour the evolution of selfing, and the ecological agents limiting outcross pollen are often not identified. Pollen discounting appears to be highly variable and important in systems where selfing involves multiple floral adaptations, yet seed discounting has rarely been investigated. Although reproductive assurance appears likely as a leading factor facilitating the evolution of selfing, studies must account for both seed and pollen discounting to adequately test this hypothesis.

Conclusions

The transmission advantage and reproductive assurance ideas describe components of gene transmission that favour selfing. Future work should move beyond their dichotomous presentation and focus upon understanding whether selection through pollen, seed or both explains the spread of selfing-rate modifiers in plant populations.     

Floral adaptation and diversification under pollen limitation

Pollen limitation (PL) of seed production creates unique conditions for reproductive adaptation by angiosperms, in part because, unlike under ovule or resource limitation, floral interactions with pollen vectors can contribute to variation in female success. Although the ecological and conservation consequences of PL have received considerable attention in recent times, its evolutionary implications are poorly appreciated. To identify general influences of PL on reproductive adaptation compared with those under other seed-production limits and their implications for evolution in altered environments, we derive a model that incorporates pollination and post-pollination aspects of PL. Because PL always favours increased ovule fertilization, even when population dynamics are not seed limited, it should pervasively influence selection on reproductive traits. Significantly, under PL the intensity of inbreeding does not determine whether outcrossing or autonomous selfing can evolve, although it can affect which response is most likely. Because the causes of PL are multifaceted in both natural and anthropogenically altered environments, the possible outcrossing solutions are diverse and context dependent, which may contribute to the extensive variety of angiosperm reproductive characteristics. Finally, the increased adaptive options available under PL may be responsible for positive global associations between it and angiosperm diversity.     

Among-species differences in pollen quality and quantity limitation: implications for endemics in biodiverse hotspots

Background and Aims

Insufficient pollination is a function of quantity and quality of pollen receipt, and the relative contribution of each to pollen limitation may vary with intrinsic plant traits and extrinsic ecological properties. Community-level studies are essential to evaluate variation across species in quality limitation under common ecological conditions. This study examined whether endemic species are more limited by pollen quantity or quality than non-endemic co-flowering species in three endemic-rich plant communities located in biodiversity hotspots of different continents (Andalusia, California and Yucatan).

Methods

Natural variations in pollen receipt and pollen tube formation were analysed for 20 insect-pollinated plants. Endemic and non-endemic species that co-flowered were paired in order to estimate and compare the quantity and quality components of pre-zygotic pollination success, obtained through piecewise regression analysis of the relationship between pollen grains and pollen tubes of naturally pollinated wilted flowers.

Key Results

Pollen tubes did not frequently exceed the number of ovules per flower. Only the combination of abundant and good quality pollen and a low number of ovules per flower conferred relief from pre-zygotic pollen limitation in the three stochastic pollination environments studied. Quality of pollen receipt was found to be as variable as quantity among study species. The relative pollination success of endemic and non-endemic species, and its quantity and quality components, was community dependent.

Conclusions

Assessing both quality and quantity of pollen receipt is key to determining the ovule fertilization potential of both endemic and widespread plants in biodiverse hotspot regions. Large natural variation among flowers of the same species in the two components and pollen tube formation deserves further analysis in order to estimate the environmental, phenotypic and intraindividual sources of variation that may affect how plants evolve to overcome this limitation in different communities worldwide.     

Effects of pollen shortage and self-pollination on seed production of an endangered tree, Magnolia stellata

BACKGROUND AND AIMS: Pollen limitation is a significant determinant of seed production, and can result from both insufficient pollen quantity (pollen shortage) and quality (mainly relating to self-pollination). For animal-pollinated tree species with large floral displays, pollen limitation may be determined by a balance between increased pollen quantity due to increased attractiveness for pollinators, countered by increased self-pollination due to increased geitonogamy. The contributions of pollen shortage and self-pollination on seed production were quantitatively examined in the natural pollination of an insect-pollinated, dichogamous, endangered tree, Magnolia stellata, which has a large, showy floral display. METHODS: Manual self- and cross-pollinations were conducted to determine the effects of selfing on seed production. The outcrossing rate was measured using microsatellite analyses of open-pollinated seeds, and the embryo mortality rate caused by self-pollination was indirectly estimated. The frequency of ovule mortality due to pollen shortage was also inferred using the embryo mortality and ovule survival rates from natural pollination. KEY RESULTS: The average fruit set, seed set per fruit, and ovule survival rate per tree from hand cross-pollination were 1.37, 3.15, and 3.34 times higher than those from hand self-pollination, respectively, indicating that self-pollination causes inbreeding depression for fruit and seed set. The multilocus-outcrossing rate (t(m)) was intermediate, 0.632, and the primary selfing rate was 0.657. This indicates that frequent geitonogamous selfing occurs. The ovule mortality rate due to pollen shortage and the embryo mortality rate due to self-pollination were estimated to be 80.8 % and 45.9 %, respectively. CONCLUSIONS: It is concluded that seed production of M. stellata is strongly limited by both pollen shortage and self-pollination. Inefficient beetle-pollination and the automimicry system via asynchronous flowering might be responsible for the high level of pollen shortage and frequent geitonogamy. This is despite a large, showy floral display and the dichogamous system of the species.     

Inbreeding and outcrossing in Yucca whipplei: consequences for the reproductive success of plant and pollinator

Unlike most pollinators, yucca moths are active pollinators of their host plants. Females lay their eggs in the flowers they pollinate, and their larvae feed solely on the resulting seeds. Previous evidence suggests that the yucca moth Tegeticula maculata avoids self-pollinating their host Yucca whipplei . Other yucca moths may self-pollinate more frequently. When pollinating, yucca moths are also reported to fly large distances between plants, bypassing neighbouring plants in the process. We experimentally verify the suggestion of Pellmyr et al . that yucca is more likely to retain fruits from self-pollination if overall fruit set is low. Thus, selection on moths to avoid self-pollinating should be density dependent. We found no evidence that mating with close neighbours resulted in inbreeding depression, thus the moth's long-distance flights between plants are yet to be explained.     

Breeding system in a population of Trigonella balansae (Leguminosae)

BACKGROUND AND AIMS: Although some taxonomic studies in the genus Trigonella have been conducted, there has been no concerted effort to study the breeding system. This paper examines the floral structure and pollination system in a population of T. balansae, an annual pasture legume. METHODS: Floral morphology, hand and vector pollination, stigma receptivity, pollen tube growth, using scanning electron and fluorescence microscopy, were conducted. KEY RESULTS: Measurements of floral structure from before to after anthesis indicates an inability for T. balansae to self-pollinate and a requirement for an external vector to effectively transfer pollen from the anthers onto the stigmas of this species. Seed set can be obtained by hand or honeybee manipulation of T. balansae flowers. CONCLUSIONS: Trigonella balansae is a self-compatible species, but which requires vectors such as honeybees to bring about pollination.     

Taxonomy and breeding system in a new species of Scrophularia L. (Scrophulariaceae) from Morocco

A new species of the genus Scrophularia, S. fontqueri Ortega-Olivencia & Devesa, is described for the Western Rif mountains in Morocco. Similarities and differences with the most closely related species (S. auriculata Loefl. ex L. and S. viciosoi Ortega-Olivencia & Devesa) are indicated, and ecological and distribution data are provided. The new species displays a high degree of polyploidy (2n = 56), and has palynological characteristics similar to those of other species in this genus. The new species shows protogynous dichogamy and produces considerable amounts of nectar in both sexual phases. Nectar secretion was maximum during the early hours of the morning. Mean longevity of the flowers was 2.66 days. Both nectar secretion and floral longevity correlated strongly with mean daytime air temperature. Pollination experiments showed that interaction with pollinators is necessary for reproduction, and gametophytic homomorphic self-incompatibility was documented. This feature was confirmed by observation of pollen tubes after hand self-pollination. The seed/ovule ratio was very low.     

The structure and function of orchid pollinaria

Cohesive masses of pollen known as pollinia have evolved independently in two plant families — Orchidaceae and Asclepiadaceae. Yet, the bilateral symmetry of orchids has allowed a greater degree of specialization in pollination systems and a much greater diversity in the morphology of pollinaria — units comprising the pollinia(um) together with accessory structures for attachment to the pollinator. Pollinaria differ in the degree of cohesion of pollen in the pollinium, which may be soft, sectile (comprised of sub-units known as massulae) or hard. A single hard pollinium may contain more than a million pollen grains, yet pollen:ovule ratios in orchids are several orders of magnitude lower than in plants with powdery pollen due to the lack of wastage during transport to the stigma. Attachment of pollinia to the pollinator is usually achieved by means of a viscidium that adheres most effectively to smooth surfaces, such as the eyes and mouthparts of insects and beaks of birds. The stalk connecting a pollinium to the viscidium may be comprised of a caudicle (sporogenous in origin) and/or a stipe (derived from vegetative tissue), or be lacking altogether. Caudicles and stipes may undergo a gradual bending movement 20 s to several hours after withdrawal from the flower, the main function of which appears to be to reduce the possibility of geitonogamous pollination. Other mechanisms that promote outcrossing and pollen export in orchids include pollen carryover (achieved by sectile or soft pollinia), temporary retention of the anther cap, protandry and self-incompatibility (rare among orchids). Pollinaria ensure that large pollen loads are deposited on the stigma, thus enabling the fertilization of the large numbers of ovules in the flowers of Orchidaceae. Pollinaria also ensure efficient removal of pollen from the anther, minimal pollen wastage during transit, and a high probability of deposition on conspecific stigmas.