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Species that propagate by sexual reproduction actively guard against the fertilization of an egg by multiple sperm (polyspermy). Flowering plants rely on pollen tubes to transport their immotile sperm to fertilize the female gametophytes inside ovules. In Arabidopsis, pollen tubes are guided by cysteine-rich chemoattractants to target the female gametophyte.sup.1,2. The FERONIA receptor kinase has a dual role in ensuring sperm delivery and blocking polyspermy.sup.3. It has previously been reported that FERONIA generates a female gametophyte environment that is required for sperm release.sup.4. Here we show that FERONIA controls several functionally linked conditions to prevent the penetration of female gametophytes by multiple pollen tubes in Arabidopsis. We demonstrate that FERONIA is crucial for maintaining de-esterified pectin at the filiform apparatus, a region of the cell wall at the entrance to the female gametophyte. Pollen tube arrival at the ovule triggers the accumulation of nitric oxide at the filiform apparatus in a process that is dependent on FERONIA and mediated by de-esterified pectin. Nitric oxide nitrosates both precursor and mature forms of the chemoattractant LURE1.sup.1, respectively blocking its secretion and interaction with its receptor, to suppress pollen tube attraction. Our results elucidate a mechanism controlled by FERONIA in which the arrival of the first pollen tube alters ovular conditions to disengage pollen tube attraction and prevent the approach and penetration of the female gametophyte by late-arriving pollen tubes, thus averting polyspermy. FERONIA prevents polyspermy in Arabidopsis by enabling pectin-stimulated nitric oxide accumulation at the filiform apparatus after the first pollen tube arrives, which disengages LURE1 chemoattraction to prevent late-arriving pollen tubes from entering the ovule., Author(s): Qiaohong Duan [sup.1] [sup.2] , Ming-Che James Liu [sup.1] [sup.3] , Daniel Kita [sup.1] [sup.4] [sup.7] , Samuel S. Jordan [sup.1] , Fang-Ling Jessica Yeh [sup.1] , Robert Yvon [...]

Environmental stress in plants impacts many biological processes, including male gametogenesis, and affects several cytological mechanisms that are strongly interrelated. To understand the likely impact of rising temperature on reproductive fitness in the climate change regime, a study of tapetal mitosis and its accompanying meiosis over seasons was made to elucidate the influence of temperature change on the cytological events occurring during microsporogenesis. For this we used two species of an environmentally sensitive plant system, i.e., genus Cymbopogon Sprengel (Poaceae), namely Cymbopogon nardus (L.) Rendle var. confertiflorus (Steud.) Bor (2n = 20) and Cymbopogon jwaruncusha (Jones) Schult. (2n = 20). Both species flower profusely during extreme summer (48°C) and mild winter (15°C) but support low and high seed fertility, respectively, in the two seasons. We have shown that tapetal mitotic patterns over seasons entail differential behavior for tapetal mitosis. During the process of tapetum development there are episodes of endomitosis that form either (i) an endopolyploid genomically imbalanced uninucleate and multinucleate tapetum, and (or) (ii) an acytokinetic multinucleate genomically balanced tapetum, with the progression of meiosis in the accompanying sporogenous tissue. The relative frequency of occurrence of the two types of tapetum mitosis patterns is significantly different in the two seasons, and it is found to be correlated with the temperature conditions. Whereas, the former (genomically imbalanced tapetum) are prevalent during the hot summer, the latter (genomically balanced tapetum) are frequent under optimal conditions. Such a differential behaviour in tapetal mitosis vis-a-vis temperature change is also correspondingly accompanied by substantial disturbances or regularity in meiotic anaphase disjunction. Both species show similar patterns. The study underpins that tapetal mitotic behaviour per se could be a reasonable indicator to elucidate the effect of climate change on reproductive fitness. Key words: tapetum mitosis, tapetum division pattern, tapetum and meiosis, Cymbopogon, aromatic grass. Les stress environnementaux chez les plantes affectent plusieurs processus biologiques dont la gametogenese male et touchent plusieurs mecanismes cytologiques qui sont etroitement relies. Afin de comprendre l'impact vraisemblable des temperatures croissantes sur le succes reproductif dans le contexte des changements climatiques, les auteurs ont etudie la meiose dans le tapetum et la meiose qui l'accompagne au cours de plusieurs saisons afin de determiner l'influence des changements de temperature sur les evenements cytologiques survenant au cours de la microsporogenese. Pour ce faire, les auteurs ont employe deux especes composant un systeme vegetal sensible aux changements environnementaux, soit le genre Cymbopogon Sprengel (Poaceae) et particulierement le Cymbopogon nardus (L.) Rendle var. confertiflorus (Steud.) Bor (2n = 20) et le Cymbopogon jwaruncusha (Jones) Schult. (2n = 20). Ces deux especes fleurissent de maniere prolifique tant durant des canicules estivales (48°C) que des froids hivernaux moderes (15°C), mais cela se traduit respectivement par une fecondite faible et elevee au cours des deux saisons. Les auteurs montrent que les mitoses au sein du tapetum se deroulent de maniere differentielle au cours des saisons. Durant le developpement du tapetum, des episodes d'endomitose surviennent pour produire : (i) un tapetum endopolyploide uninuclee/multinuclee presentant un desequilibre genomique ou (ii) une mitose acytocinetique menant a un tapetum multinuclee mais equilibre en ce qui a trait a son genome et la poursuite de la meiose dans les tissus sporogenes voisins. La frequence relative des deux deroulements de la mitose differait significativement selon la saison et etait correlee avec la temperature. Tandis que le tapetum presentant un desequilibre genomique etait predominant durant les conditions estivales chaudes, le tapetum balance etait plus frequent sous des conditions optimales. Une telle difference de comportement dans la mitose au sein du tapetum en fonction de la temperature est egalement accompagnee de perturbations importantes ou de regularite de la disjonction lors de l'anaphase meiotique. Les deux especes presentent des changements similaires. Cette etude illustre que le comportement mitotique au sein du tapetum serait en soi un indicateur raisonnable pour determiner l'effet des changements climatiques sur la reproduction. [Traduit par la Redaction] Mots-cles : mitose au sein de tapetum, mode de division dans le tapetum, tapetum et meiose, Cymbopogon, graminee aromatique., Introduction The genus Cymbopogon Sprengel comprises a species complex of aromatic grasses that is a source of valuable essential oil used in perfumery and aromatherapy (Lavania 2000; Lavania et al. [...]

Plant species diversity in Eurasian wetlands and grasslands depends not only on productivity but also on the relative availability of nutrients, particularly of nitrogen and phosphorus (1-4). Here we show that the impacts of nitrogen:phosphorus stoichiometry on plant species richness can be explained by selected plant life-history traits, notably by plant investments in growth versus reproduction. In 599 Eurasian sites with herbaceous vegetation we examined the relationship between the local nutrient conditions and community-mean life-history traits. We found that compared with plants in nitrogen-limited communities, plants in phosphorus-limited communities invest little in sexual reproduction (for example, less investment in seed, shorter flowering period, longer lifespan) and have conservative leaf economy traits (that is, a low specific leaf area and a high leaf dry-matter content). Endangered species were more frequent in phosphorus-limited ecosystems and they too invested little in sexual reproduction. The results provide new insight into how plant adaptations to nutrient conditions can drive the distribution of plant species in natural ecosystems and can account for the vulnerability of endangered species., Species diversity is influenced both by overall nutrient availability and by nutrient stoichiometry--that is, how the ratio of available nutrients relates to their consumers' requirements (5). In terrestrial plant communities [...]

The growth of pollen tubes is one of the most characteristic events in angiosperm reproduction. This article describes an activity for visualizing the journey and guidance of pollen tubes in the reproductive structures of a flowering plant. The activity uses a semi-in vivo system with rapid-cycling Brassica rapa, also known as Fast Plants. Isolated ovules were used to attract pollen tubes that were triggered to grow through explants of female flower parts. The activity provides insight into the in vivo situation of plant reproduction, appealing visual results, and the development of science process skills. Key Words: Fast Plants; Brassica rapa; pollen germination; guidance; flower anatomy., Sexual reproduction is one of the most important features in the life cycle of higher plants. In contrast to animals, plant fertilization is based on the interplay between multicellular haploid [...]

Sexual reproduction in non-vascular plants requires unicellular free-motile sperm to travel from male to female reproductive structures across the terrestrial landscape (1). Recent data suggest that microarthropods can disperse sperm [...]

* Premise of the study: Species in the aquatic genus Nymphoides have inflorescences that appear to arise from the petioles of floating leaves. The inflorescence-floating leaf complex can produce vegetative propagules and/or additional inflorescences and leaves. We analyzed the morphology of N. aquatica to determine how this complex relates to whole plant architecture and whether whole plant growth is sympodial or monopodial. * Methods: We used dissections, measurements, and microscopic observations of field-collected plants and plants cultivated for 2 years in outdoor tanks in south Florida, USA. * Key results: Nymphoides aquatica had a submerged plagiotropic rhizome that produced floating leaves in an alternate/spiral phyllotaxy. Rhizomes were composed of successive sympodial units that varied in the number of leaves produced before the apex terminated. The basic sympodial unit had a prophyll that subtended a renewal-shoot bud, a short-petioled leaf (SPL) with floating lamina, and an inflorescence; the SPL axillary bud expanded as a vegetative propagule. Plants produced either successive basic sympodial units or expanded sympodia that intercalated long-petioled leaves between the prophyll and the SPL. * Conclusions: Nymphoides aquatica grows sympodially, forming a rhizome composed of successive basic sympodia and expanded sympodial units. Variations on these types of sympodial growth help explain the branching patterns and leaf morphologies described for other Nymphoides species. Monitoring how these two sympodial phases are affected by water depth provides an ecologically meaningful way to assess N. aquatica's responses to altered hydrology. Key words: aquatic plant; clonal propagation; Everglades; heterophylly; leaf dimorphism; prophyll; slough; sympodial; vegetative reproduction. doi: 10.3732/ajb.1000242

* Premise of the study: Pyrus callervana Decne., an ornamental tree species introduced from China, is a relatively new invasive that has only recently begun to spread across the United States after intraspecific hybridization between cultivars. The function of such hybridization in the evolution of invasiveness is still relatively understudied, especially with respect to the initial establishment and persistence of invasive genotypes. Multiple introductions of genetically divergent populations or cultivars may benefit from new genetic combinations created during hybridization events and/or release from Allee effects in founder populations. * Methods: We quantified the outcome of intraspecific hybridization between cultivars of P. calleryana in a common garden. Measures of the reproductive success and establishment ability of their early- and advanced-generation hybrid offspring were collected to assess the likelihood of particular cultivar genotypes to establish in invasive populations. These traits also were compared between cultivated and invasive parents to identify any generational differences in invasive potential. * Key results: Differences were detected in measures of reproductive ability, but no group emerged as consistently more fecund. Advanced-generation hybrids also had significantly less biomass, indicating a reduction in hybrid performance relative to that of the cultivated progeny. * Conclusion: Ultimately, this study indicates that increased spread of P. calleryana has been initiated by introduction of multiple cultivar types and subsequent widespread planting and is not the result of an inherent fitness advantage of hybrid progeny. Key words: cultivation; intraspecific hybridization; invasive; propagule pressure; Pyrus calleryana. doi: 10.3732/ajb.1000113

* Premise of the study: The evolution of asexual seed production (apomixis) from sexual relatives is a great enigma of plant biology. The genus Boechera is ideal for studying apomixis because of its close relation to Arabidopsis and the occurrence of sexual and apomictic species at low ploidy levels (diploid and triploid). Apomixis is characterized by three components: unreduced embryo-sac formation (apomeiosis), fertilization-independent embryogenesis (parthenogenesis), and functional endosperm formation (pseudogamy or autonomous endosperm formation). Understanding the variation in these traits within and between species has been hindered by the laborious histological analyses required to analyze large numbers of samples. * Methods: To quantify variability for the different components of apomictic seed development, we developed a high-throughput flow cytomelric seed screen technique to measure embryo: endosperm ploidy in over 22000 single seeds derived from 71 accessions of diploid and triploid Boechera. * Key results: Three interrelated features were identified within and among Boechera species: (1) variation for most traits associated with apomictic seed formation, (2) three levels of apomeiosis expression (low, high, obligate), and (3) correlations between apomeiosis and parthenogenesis/pseudogamy. * Conclusions: The data presented here provide a framework for choosing specific genotypes for correlations with large 'omics' data sets being collected for Boechera to study population structure, gene flow, and evolution of specific traits. We hypothesize that low levels of apomeiosis represent an ancestral condition of Boechera, whereas high apomeiosis levels may have been induced by global gene regulatory changes associated with hybridization. Key words: apomeiosis; apomixis; Boechera; embryo; endosperm; flow cytometry; parthenogenesis; pseudogamy. doi: 10.3732/ajb.1000188

* Premise of the study: Habitat fragmentation is often assumed to result in limited genetic diversity across impacted plant communities. Central Florida has undergone extensive anthropogenic changes, while also harboring large numbers of endemic species. In this study, we assessed genetic structure and dependence on clonality in a central Florida endemic, Illicium parviflorum (Illiciaceae), as well as evaluated genetic diversity of this species in horticultural stocks. * Methods: Six sites were sampled across the geographic range of I. parviflorum. A PCR-based assay using intersimple sequence repeats (ISSRs) was used to assess genetic structure. * Key results: Results, based on 26 ISSR loci, suggest that clonal structure plays a role in all populations, with PD values ranging from 0.25 to 0.50. Only two populations exhibited unique genotypes, while the remaining four populations shared genotypes. Horticultural samples all shared one genotype, which can be traced back to a single natural population. * Conclusions: Clonal reproduction is an important factor in the maintenance of natural populations of I. parviflorum, although the degree to which this is true varies by population. Horticultural samples likely represent a single or very few collection events, indicating the need for greater genetic diversity within horticultural stocks. Further analyses using microsatellites are planned to confirm these results. doi: 10.3732/ajb.0900291

* Premise of the study: Flowering plants that rely on pollinators for most of their reproduction may experience unpredictable and inconsistent availability of effective pollinators throughout their reproductive lifetime. We investigated the reproductive ecology of two subspecies of the tufted evening primrose, Oenothera cespitosa, which occupy geographically and edaphically distinct habitats in western North America: O. cespitosa subsp, navajoensis inhabits sandstone soils on open sites or rocky slopes in the Colorado Plateau and O. cespitosa subsp, cespitosa grows in clay soils on talus slopes and exposed rocky ridges in the western Great Plains and northern Rocky Mountains of the United States. * Methods: Pollen augmentation and selfing experiments, floral visitor observations, and single-visit effectiveness experiments were conducted over 4 years to examine the breeding system and spatiotemporal variation in pollinator behavior, assemblage, and abundance at different populations for each subspecies. * Key results: Both subspecies of O. cespitosa were self-incompatible and pollen-limited, suggesting that the relative abundance, effectiveness, and movement patterns of different insects as pollinators influenced the quality and quantity of seed production in these plants. Medium-sized vespertine hawkmoths (Hyles lineata, Sphinx vashti) were effective pollinators when present, as were large matinal bees (Anthophora affabilis, A. dammersi, Xylocopa tabaniformis androleuca), whereas small oligolectic Lasioglossum bees primarily functioned as pollen thieves in the evening and morning. * Conclusions: These findings highlight the importance of variability of pollinator composition and abundance in the evolution of plant breeding systems and reproductive success at varying spatial and temporal scales. Key words: bees; breeding system; hawkmoths; Oenothera cespitosa; Onagraceae; plant reproductive success; pollination; spatiotemporal variation. doi: 10.3732/ajb.1000086

* Premise of the study: Current reproduction in polycarpic plants may be affected by a wide variety of factors, including carryover or 'historical' effects derived from environments experienced early in life and from previous investments in organ preformation or other life functions, such as growth or reproduction. Historical effects as determinants of plant reproductive success in a specific season have received considerably less attention than events during the current reproductive episode, especially for short-lived plants. * Methods: We used structural equation modeling to assess direct and indirect relations between past reproduction and both subsequent reproduction and growth in Helianthemum squamatum, a short-lived polycarpic plant. Additionally, we explored the effects of current reproduction on future survival (binomial variable) by using logistic regression. * Key results: Historical effects derived from previous growth positively affected current reproduction, suggesting the existence of a reproductive hierarchy in the population, in which some individuals are consistently better growers and seeders. The lack of effects of current reproduction on future reproduction, together with the existence of a negative effect on future survival, suggests that to invest maximally in reproduction at the expense of a short life span may be the optimal strategy for H. squamatum, a species inhabiting very unpredictable ecosystems. * Conclusions: Historical effects derived from past investments in growth or reproduction may have important consequences for current plant performance and may play an essential role in shaping life histories. Key words: Cistaceae; Helianthemum squamatum; Mediterranean; reproductive hierarchy; reproductive lifetime; semiarid; structural equation modeling. doi: 10.3732/ajb.1000072

Androgenesis can be defined as the set of biological processes leading to an individual genetically coming exclusively from a male nucleus. Androgenesis was traditionally considered as the spontaneous, in vivo development of a male-derived haploid embryo from a fertilized egg where the female nucleus is eliminated. However, at present it is also possible to generate androgenic haploids/doubled haploids through in vitro microspore embryogenesis and by in vitro meiocyte-derived callogenesis. These three androgenic alternatives clearly differ in the inducible stage, but lead to the same final haploid or doubled haploid product. Whereas microspore embryogenesis is widely studied and applied, the other two routes are much less known. In this paper, the evidence accounting for the existence of these three alternative pathways is revised, as well as the mechanisms potentially involved in their induction. Their differences and similarities are discussed from a biological perspective, leading to the notion that they might represent an ancient survival mechanism triggered by similar factors. La androgenesis se define como el conjunto de vias biologicas que dan lugar a un individuo cuyo fondo genetico proviene exclusivamente de un nucleo de origen masculino. Tradicionalmente, el concepto de androgenesis estaba restringido al desarrollo espontaneo, in vivo, de un embrion haploide o doble haploide a partir de una celula huevo fecundada en la que el material gendtico de origen femenino era inactivado o eliminado. Sin embargo, hoy en dia sabemos que existen otras vias para conseguir haploides o doble haploides androgenicos, mediante embriogenesis de microsporas y mediante la formacion de callos derivados de meiocitos. Estas tres alternativas androgdnicas difieren claramente en la etapa en la que es posible la induccion, pero dan lugar al mismo producto haploide o doble haploide final. Mientras que la embriogenesis de microsporas es un fenomeno ampliamente estudiado y de clara aplicacion practica, las otras dos rutas son mucho menos conocidas. En este trabajo se revisan las evidencias existentes al respecto de estas tres alternativas androgenicas, asi como los mecanismos potencialmente implicados en su induccion. Tambien se discuten sus diferencias y semejanzas, desde un punto de vista biologico, para llegar a la hipotesis de que estas rutas podrian representar un mecanismo atfivico de supervivencia activado por factores similares. Keywords Microspore Embryogenesis * Meiocyte-Derived Callogenesis * Male-Specific Parthenogenesis * Plant Reproduction * Haploid * Doubled Haploid, Introduction During evolution, plant cells have retained the capacity to express virtually any part of their coding genome. This remarkable feature, called totipotency, allows for a differentiated cell to dedifferentiate [...]

Premise of the study: The study of geographic variation in ecologically important traits within and among taxa is a first step toward understanding the environmental factors that contribute to population differentiation and species divergence. This study examines variation in mean sex allocation per flower (androecium mass/gynoecium mass) among 49 wild populations representing 12 Pedicularis species across an elevation gradient on the eastern Tibetan Plateau. Methods: We used population means to evaluate sources of variation in per-flower sex allocation within and across species. In particular, we evaluate the relative influence of intrinsic (i.e., plant size, estimated as aboveground stem biomass) vs. extrinsic factors affecting mean sex allocation among populations. Key results: Mean sex allocation per flower (the relative investment in male floral organs) is negatively correlated with mean plant size; populations of large plants produce relatively female-biased flowers. This relationship between mean plant size and mean sex allocation is not statistically significant, however, when the effect of elevation is controlled statistically. Among populations within and across species, mean sex allocation increases with elevation. This relationship persists even when the effect of mean plant size is controlled statistically. Factors associated with increasing elevation appear to favor genotypes and/ or taxa with male-biased flowers. Conclusion: Extrinsic environmental conditions may be more important than intrinsic resource status in determining patterns of geographic variation in mean sex allocation among populations or species of Pedicularis. We cannot conclude whether the effect of elevation on mean sex allocation is the result of environmentally induced plasticity, genetically based adaptation, or species sorting, but it is only partly mediated by mean plant size. Key words: elevation; geographic variation; Orobanchaceae; Pedicularis; population differentiation; sex allocation. doi: 10.3732/ajb.0900301

* Premise of the study: Embryonic inbreeding depression is a key influence on mating system evolution and can be difficult to estimate in self-incompatible species. A pollen chase experiment was used to estimate the magnitude of embryonic inbreeding depression in Costa Rican Witheringia solanacea, a species polymorphic for self-incompatibility (SI). In a pollen chase experiment, bud self-pollinations are followed after anthesis by outcross pollinations, with a comparable pair of outcross pollinations used as a control. Lowered seed set for the self-precedence treatment indicates embryonic inbreeding depression. * Methods: Embryonic inbreeding depression was assayed for self-compatible (SC) individuals and for SI plants from two populations that differ quantitatively in the onset and enzymatic activity of their SI response. Microsatellite markers were used to assay the selfing rate of a sample of surviving progeny from the prior self-pollination treatment. * Key results: SC individuals showed no evidence of embryonic inbreeding depression. In SI plants, prior self- pollination reduced seed number by 28-70%, depending on population. Microsatellite genotyping revealed that embryonic inbreeding depression was even more severe than estimated by the phenotypic data: for mature fruits resulting from self- pollination precedence, the majority of the progeny were the result of outcross fertilization. * Conclusions: Lineage-specific purging of recessive lethals has accompanied the evolution of SC in this species. SI populations show contrasting levels of embryonic inbreeding depression, with nearly complete embryonic lethality upon selfing in the Monteverde population. In the lace of high embryonic inbreeding depression, an increase in selling rate can evidently occur only under severe pollen limitation. Key words: Costa Rica: embryonic inbreeding depression; evolution; mating system: self-compatibility: self- incompatibility; Solanaceae; Witheringia solanacea. doi: 10.3732/ajb.1000020

* Premise of the study: A mixture of outcrossing and selfing is often observed in plant populations. Although mixed mating is ubiquitous, it has several potential evolutionary explanations. Mixed mating may be actively maintained by selection, passively determined by the pollination environment, or a transitional stage during the evolution of self-fertilization. * Methods: We studied patterns of self-compatibility and selfing rates in a population of Leavenworthia alubamica that recently lost self-incompatibility. We also experimentally tested whether natural selection against selfing at the pre-or postzygotic stage is sufficient to explain mixed mating in this population. * Key results: Visualizing pollen tube growth following self-pollination, we found that nearly all plants were fully self-compatible. Progeny array analysis revealed that the average selfing rate of the population was s = 0.523. The inbreeding coefficient in the parents (F = 0.539) exceeded the amount expected if the selfing rate (s) were constant [[F.sub.cq] = s/(2 - s)l, indicating either population subdivision or higher selfing rates in the past. Inference of family-level selfing rates revealed substantial variation. Experiments found that self and outcross pollen fertilized nearly equal numbers of ovules in competition. Comparison of seed production following self-or cross-pollination failed to implicate early acting inbreeding depression as a factor maintaining mixed mating. * Conclusions: The results of our experiments suggest that mixed mating is not maintained by selection against self- pollen or zygotes in this population. Mixed mating is most likely a byproduct of the pollination process but may also be a transitional stage during the evolution of higher selfing rates. Key words: bet hedging; Brassicaceae; fluctuating environments; inbreeding depression; Leavenworthia alubamica; mating system; pollen competition; reproductive assurance. doi: 10.3732/ajb.1000032

The reproductive system is one of the key mechanisms that determine genetic diversity at different biological levels. However, few attempts have been made to assess the consequences of asexual reproduction by comparing genetic structure and fecundity of seed families in asexually and sexually reproducing individuals. We have examined two similar hawthorn species, Crataegus crusgalli and C. punctata, that differ in ploidy level and breeding system. Fecundity (per-fruit seed set) and microsatellite genotypes for five loci were determined in 18 and 26 seed families of C. crus-galli and C. punctata (totals of 83 and 118 embryos), respectively. Compared with the sexual diploid C. punctata, tetraploid C. crus-galli shows similar fecundity, but lower genotypic diversity within and between seed families. Reproduction in the tetraploid, while predominantly apomictic, is nevertheless accompanied by outcrossing and selfing. We conclude that in C. crus-galli pollen flow between conspecific individuals is limited, and the combination of pollen fertility, self-compatibility, and pseudogamous apomixis provides reproductive assurance in these tetraploids. Reproductive assurance, in turn, may explain the derived floral architecture seen in most North American tetraploid hawthorns. We also discuss analytical approaches for inferring mating-system parameters in tetraploids and for comparing microsatellite variation across ploidy levels. Key words: Apomixis; family genetic structure; fecundity; embryo genotypes; microsatellite; outcrossing; pollen flow; polyploidy; progeny array; Rosaceae. doi: 10.3732/ajb.0900091

Viability of long-distance pollen links ecological models to the genetic structure of forest tree populations, determining how forests will adapt to climate change and how far genes flow from genetically modified (GM) pine plantations. Addressing this landscape-scale inquiry is feasible when the pollen source, the delivery system, and the receiver field can be made explicit. To this end, I measured long-distance pollen germination along a 160-km transect along the North Carolina coastline, including 45 000 ha of mature Pinus taeda plantations and barrier islands. Using this system, I tested three hypotheses: (1) pine pollen germinates 'after dispersal on meso-scale distances, (2) sodium chloride exposure reduces germination of pollen captured over open saltwater, and (3) viable pine pollen is present at high altitudes before local peak pollen shed. The experimental findings are as follows: pine pollen had germination rates of 2 to 57% after dispersal at distances from 3 to 41 kin, sodium chloride solutions mildly reduced P taeda pollen germination, and viable pine pollen grains were captured at an altitude of 610 m. GM pine plantings thus have a potential to disperse viable pollen at least 41 km from the source. Wind and rainfall, as integral parts of regional atmospheric systems, together exert a powerful influence on the genetic structure of forest tree populations. Key words: aerosols; climate change; conifer reproductive biology; genetically modified (GM) forest trees; heterospory; long-distance dispersal; male gametophyte; North Carolina; Pinaceae; Pinus taeda; U.S. Forest Health Initiative. doi: 10.3732/ajb.0900255

Flowering sex ratios of d.ioecious plants are commonly male-biased but rarely female-biased. While greater costs of reproduction from females have been repeatedly demonstrated and explain male biases, male reproductive costs almost never exceed female costs, making the origins of female biases enigmatic. I investigated the seagrasses Phyllospadix scouleri and P. serrulatus (surfgrasses), which have some of the most extreme female-biased sex ratios documented (>90% female), to identify the mechanisms driving sex ratio bias. I developed sex-linked amplified fragment length polymorphism (AFLP) markers and applied them to three P. scouleri life stages at four sites to determine when during the life cycle sex ratio bias arises. Sex ratios were even among seedlings but became more female-biased at later life stages, indicating that sex ratios were driven by male-biased mortality. To identify when during the life cycle sex ratio bias developed, I examined sex differences in survival among seedlings and three aspects of reproductive costs that could potentially generate biased sex ratios under field conditions. No differences in seedling survival between the sexes were detected, and there was no evidence of substantial sex differences in costs of reproduction. I found no support for a trade-off between current and future reproduction or between reproductive investment and growth. Thus, costs of reproduction appear unlikely to drive sex ratio bias in surfgrass. Instead, small sex differences in growth and survival spread across the life cycle appear to be responsible for female-biased sex ratios and suggest that life history trade-offs other than reproductive costs drive sex ratio bias. Key words: cost of reproduction; dioecy; marine angiosperm; Olympic Peninsula, Washington, USA; Phyllospadix scouleri; Phyllospadix serrulatus; seagrass; seedling survivorship; sex-linked genetic marker; sex ratio.

Reproductive barriers play a major role in the origin and maintenance of biodiversity by restricting gene flow between species. Although both pre- and postzygotic barriers often isolate species, prezygotic barriers are thought to contribute more to reproductive isolation. We investigated possible reproductive barriers between Leavenworthia alabamica and L. crassa, parapatric species with high morphological and ecological similarity and the ability to hybridize. Using greenhouse and field experiments, we tested for habitat isolation and genetic incompatibilities. From controlled crosses, we identified unilateral incompatibility (a partial prezygotic barrier associated with the self-incompatibility system), but no evidence of other genetic incompatibilities. We found a small reduction in pollen viability of F1 hybrids and early germination of [F.sub.1], [F.sub.2], and BC hybrids relative to L. alabamica and L. crassa in a common garden experiment, but the effect on fitness was not tested. Field studies of hybrid pollen viability and germination are needed to determine if they contribute to reproductive isolation. In a reciprocal transplant, we found no evidence of habitat isolation or reduced hybrid survival (from seedling to adult stage) or reproduction. These data suggest unilateral incompatibility partially reproductively isolates L. alabamica and L. crassa, but no other reproductive barriers could be detected. Key words: Brassicaceae; genetic incompatibilities; hybrid fitness; habitat isolation; Leavenworthia alabamica; Leavenworthia crassa; reproductive barriers; unilateral incompatibility. doi: 10.3732/ajb.0900184

Theory suggests that iteroparity may confer greater fitness than semelparity in situations in which temporal environmental variation is high and unpredictable. Variable age-specific mortality, density dependence, and other factors may also favor iteroparity over semelparity. Here, we empirically test the adaptive benefits of greater numbers of reproductive years in a study of reproductive schedules in an experimental population of a short-lived polycarpic perennial, Plantago lanceolata. A large experimental population was established that included four cohorts with similar genetic structure. Individuals were censused for mortality, size, and reproduction for seven years. Plants experienced variable numbers of reproductive years, but one or two years were most common (~46.7% of the population reproduced only once). The probability of flowering at least once prior to death was determined strongly by extrinsic, environmental or intrinsic but environmentally influenced variables, including early-life size, cohort, and block, but also varied with a number of interactions involving paternal lineage. Maternal effects explained small but significant components of the variance in the number of reproductive years among individuals in each cohort, while paternal effects were significant in only two cohorts. Number of reproductive years contributed significantly to fitness in this system, more so than all other variables tested, although most of the variation in relative fitness may be attributed ultimately to environmental influences. We suggest that the high proportion of each cohort composed of plants reproducing only once may be due to environmental constraints on either growth or size. Such environmental influences, particularly on early life size, may result in small but important indirect effects on fitness. Key words: inflorescence; iteroparity; Plantago lanceolata; reproductive schedule; reproductive timing; semelparity; Virginia, USA.

This paper explores the relative impacts of reduction and polymerization on the evolution of reproductive structures in the small but morphologically diverse family Centrolepidaceae. Centrolepidaceae are closely related to Restionaceae and belong to the large order Poales, which also includes the grasses. In the largest genus of Centrolepidaceae, Centrolepis, the reproductive structures are viewed either as highly unusual aggregations of reduced flowers (the pseudanthial interpretation) or as unique flowers evolved through extreme reduction in the androecium, usually accompanied by a drastic increase in carpel number and elaboration of the entire gynoecium. Comparative data are here presented on gynoecia of all three genera of Centrolepidaceae; these data strongly support the latter (euanthial) interpretation. The combined phenomenon of carpel multiplication and decrease in stamen number is unexpected in a predominantly wind-pollinated lineage. Applying a pseudanthial interpretation would create a considerable morphological gap with reproductive structures of other Poales, whereas accepting a euanthial concept allows an almost continuous morphological series with related taxa. Key words: anatomy; Centrolepidaceae; development; flower; gynoecium; inflorescence; morphology; pseudanthium; Poales. doi: 10.3732/ajb.0900074

The cost of sexual reproduction is incurred when the current reproductive episode contributes to a a decline in future plant performance. To test the hypotheses that a trade-off exists between current sexual reproduction and subsequent clonal regeneration and that resources limit reproduction and regeneration, plants of the widespread moss Pterygoneurum ovatum were subjected to induced sporophytic abortion, upper leaf removal, and nutrient amendment treatments. Sexually reproducing plants were slower or less likely to produce regenerative structures (protonemata or shoots) and produced fewer regenerative tissue areas or structures. The ability and the timeline to reproduce sexually and regenerate clonally were unaffected by an inorganic nutrient amendment. However, when leaves subtending the sporophyte were removed, the sporophytes were less likely to mature, tended to take a longer time to mature, and were smaller compared to sporophytes from shoots with a full complement of upper leaves. Our findings indicate that plants investing in sexual reproduction suffer a cost of decreased clonal regeneration and indicate that sporophyte maturation is resource-limited, with upper leaves contributing to the nutrition of the sporophyte. This study represents only the second explicit experimental demonstration of a trade-off between sexual and asexual reproduction in bryophytes. Key words: abortion; bryophyte; Pottiaceae; protonema; Pterygoneurum ovatum; regeneration; sexual reproduction; sporophyte; trade-offs. doi: 10.3732/ajb.0900084

To clarify the budding pattern of Wickerhamomyces pijperi, the vegetative cells were observed by scanning electron microscopy. The cells grew by bipolar budding, but cells that budded from the shoulder of a mother cell were occasionally observed. We examined the cell morphology and phylogeny of five strains of Wickerhamomyces sp. isolated in Thailand as well as seven W. pijperi and three Wickerhamomyces sp. strains that were preserved in culture collections. Phylogenetic analysis based on three different nucleotide sequences (D1/D2 domain of 26S rDNA, the actin gene ACT1 and the elongation factor 2 gene EF2) indicated that all the strains belonged to the genus Wickerhamomyces and were neighbours of the type strain W. pijperi NBRC [1290.sup.T]. The strains fell into two groups in this analysis. The budding patterns of the strains were carefully observed by staining the bud scars, and these patterns were categorized into three groups: types I-III. Type I included cells that grew by bipolar budding and formed multiple scars, type III included cells that grew by multilateral budding and formed a single scar, and type II included cells that exhibited a mixture of type I and type III patterns. Among the 15 strains, 12 strains, including W. pijperi NBRC [1290.sup.T], mainly exhibited type I or type II budding patterns; these strains belonged to group 1 of the phylogenetic analysis. The remaining three strains, which belonged to group 2, exhibited either type II or type III patterns. Thus the phylogenetic relationship and budding patterns are related. Moreover, some cells also exhibited budding characteristics that were intermediate between bipolar and multilateral budding.

Studies of mycoheterotrophs, defined as plants that obtain carbon resources from associated mycorrhizal fungi, have fundamentally contributed to our understanding of the importance and complexity of symbiotic ecological interactions. However, to date, the reproductive ecology of these organisms remains empirically understudied, with existing literature presenting hypotheses about traits including a generalist pollination syndrome and autogamous self-pollination. To address this gap in our knowledge of the reproductive ecology of mycoheterotrophic plants, we comparatively analyzed three species of two monotropoid genera, Monotropa and Monotropsis. During three consecutive years of field observations and manipulations of four populations of Monotropa uniflora, seven of M. hypopitys (both red and yellow color forms), and two of Monotropsis odorata, we investigated flowering phenology, pollination ecology, breeding system, floral herbivory, and reproductive effort and output. Contrary to previous predictions, our results revealed that taxa are largely outcross-pollinated and specialized toward Bombus pollinators. Additionally, species differ in breeding system, timing and duration of reproductive development, fluctuations in reproductive effort and output, and fitness impacts of herbivory. This study is the first thorough investigation of the reproductive ecology of mycoheterotrophic species and provides insight into possible limitations in reproductive traits imposed by a mycoheterotrophic life history. Key words: breeding system; Ericaceae; Monotropa; Monotropsis; mycoheterotroph; phenology; pollination; reproductive ecology. doi: 10.3732/ajb.0800319

Development of theory has outpaced experimental tests for most maintenance of diversity mechanisms. Here we demonstrate how data from biodiversity--ecosystem functioning experiments can be used to determine the mechanisms that maintain plant species diversity. We hypothesized that grassland plant diversity is maintained by two classes of mechanisms: (1) equalizing mechanisms, which reduce asymmetric competition by reducing differences in monoculture biomass production among species in mixture, and (2) species interaction mechanisms, which increase overyielding by increasing niche partitioning and facilitation among species in mixture. Specifically, equalizing mechanisms reduce the coefficient of variation in monoculture biomass production among species in mixture. Species interaction mechanisms increase species overyielding in mixture, especially for low-biomass species. We tested these predictions with a seven-year data set from an experiment that varied grassland plant species evenness and richness. We used path analysis to model effects of these mechanisms on annual and multiyear changes in diversity. We found that diversity was frequently maintained by species interaction mechanisms and was infrequently maintained by equalizing mechanisms. Species interaction mechanisms maintained diversity by allowing the species that produced the least biomass in monoculture to benefit the most from species interactions in mixture. Equalizing mechanisms infrequently maintained diversity because asymmetric competition infrequently resulted in competitive exclusion. We propose that this mechanistic framework be used to better understand the specific processes that influence diversity. Key words: biodiversity; coexistence; complementarity; ecosystem function; net biodiversity effect; productivity.

The number of ovules per flower varies over several orders of magnitude among angiosperms. Here we consider evidence that stochastic uncertainty in pollen receipt and ovule fertilization has been a selective factor in the evolution of ovule number per flower. We hypothesize that stochastic variation in floral mating success creates an advantage to producing many ovules per flower because a plant will often gain more fitness from occasional abundant seed production in randomly successful flowers than it loses in resource commitment to less successful flowers. Greater statistical dispersion in pollination and fertilization among flowers increases the frequency of windfall success, which should increase the strength of selection for greater ovule number per flower. We therefore looked for evidence of a positive relationship between ovule number per flower and the statistical dispersion of pollen receipt or seed number per flower in a comparative analysis involving 187 angiosperm species. We found strong evidence of such a relationship. Our results support the hypothesis that unpredictable variation in mating success at the floral level has been a factor in the evolution of ovule packaging in angiosperms. Key words: bet hedging; ovule; pollination; reproductive evolution; seed set; stigmatic pollen load. doi: 10.3732/ajb.0800183

Opuntia robusta has hermaphroditic, dioecious, and trioecious populations. To enhance our understanding of this breeding system diversity, we compared the reproductive output of males, females, and hermaphrodites in a trioecious population using field evaluations, controlled crosses, and progeny tests. Unisexuals were fully sterile in one sex function. Hermaphrodites were fully fertile for both functions. Consistent with the sex-allocation theory, unisexuality increased the quality and quantity (in males) of the gametes of the functional sex, relative to those of hermaphrodites, probably explained by maternal and paternal effects. The increase was higher in males than in females, suggesting a more expensive female function. Theoretically, this disproportional increase is required for unisexuals to invade a hermaphroditic population with prior selfing, negligible pollen discounting, and undetectable inbreeding depression, features found in O. robusta, therefore helping to explain dioecious populations. However, in the study population, the actual seed output of females was lower and had a higher variance than that of hermaphrodites, which also reproduce through pollen. Unisexuals are unlikely to be maintained by their actual reproductive output in this pollen-limited environment. Hermaphrodites may persist in this population by producing their seeds autonomously and by reducing interspecific fertilization by prior selfing and ovule discounting. Key words: cactus; dioecy; hermaphroditism; inbreeding depression; maternal effects; ovule discounting; pollen discounting; pollen limitation: pollen--ovule ratio; prior selfing; selfing; sex allocation. doi: 10.3732/ajb.0800301

Multiple field populations of two pairs of diploid sister taxa with contrasting mating systems in the genus Clarkia (Onagraceae) were surveyed to test predictions concerning the effects of resource status, estimated as plant size, on pollen and ovule production and on the pollen:ovule (P:O) ratio of flowers. Most theoretical models of size-dependent sex allocation predict that, in outcrossing populations, larger plants should allocate more resources to female function. Lower P:O ratios in larger plants compared to smaller plants have been interpreted as supporting this prediction. In contrast, we predicted that P:O ratio should not vary with plant size in predominantly selfing plants, in which each flower contributes to reproductive success equally through male and female function. We found that, in all four taxa, both ovule and pollen production per flower usually increased significantly with plant size and that the shape of this relationship was decelerating. However, ovule production either decelerated more rapidly than or at the same rate as pollen production with plant size. Consequently,the P:O ratio increased or had no relationship with plant size. This relationship was population-specifc (not taxon-specific) and independent of the mating system. Possible explanations for the increasing maleness with plant size are discussed. Key words: Clarkia: gametophyte production; mating system; Onagraceae; pollen to ovule ratio; size-dependent sex allocation.

A remarkably diverse Lower Cretaceous (Valanginian-Hauterivian) flora at Apple Bay, Vancouver Island, preserves seed plants at an important time of floristic evolutionary transition, about the same time as the earliest flowering plant megafossils. The fossils are permineralized in carbonate concretions and include tetrahedral seeds within cupule- or carpel-like structures. These enclosing structures, composed of elongate sclerenchyma cells with spiral thickenings that grade externally to a few layers of parenchyma, are vascularized by one collateral vascular bundle and lack trichomes. They apparently broke open to release the tightly enclosed seeds by valves. Seeds are similar to those of the Triassic seed fern Petriellaea, but are about 100 million years younger and differ in size, vascularization, integumentary anatomy, seed attachment, and number of seeds/cupule. These new seeds are described as Doylea tetrahedrasperma gen. et sp. nov., tentatively assigned to Corystospermales. Inverted cupules are reminiscent of an outer angiosperm integument rather than a carpel. Like fruits, cupules opened to release seeds at maturity, thereby foretelling several aspects of angiospermy. They show that nearly total ovule enclosure, a level of organization approaching angiospermy, was achieved by advanced seed ferns during the Mesozoic. Key words: angiospermy; corystosperms; Cretaceous; cupule; Petriellaea; seed ferns.