High male reproductive success of hermaphrodites in the androdioecious Phillyrea angustifolia

Androdioecy, the coexistence of males and hermaphrodites within a population, is a rare breeding system, often considered as unlikely to evolve because of restrictive conditions for its maintenance. Phillyrea angustifolia, a wind-pollinated shrub, is one of the handful species reported to be androdioecious. Our previous studies have shown that natural populations of this species in southern France exhibit higher male frequencies (approximately 50%) than predicted on theoretical grounds. Thus, the male functionality of hermaphrodites is still debated. To assess the functional breeding system of this species in the wild, a paternity analysis was performed with two highly polymorphic microsatellite loci on 729 seeds collected on 10 maternal shrubs in a natural population of 24 mature individuals of P. angustifolia. A large proportion of seeds were found to have been sired by pollen from outside the population. Analysis of seeds sired by individuals within the study population revealed a high male fertility of hermaphrodites resulting in a low male advantage in fertility for male plants. Intermate distances were found to have a strong impact on male reproductive success, whereas sexual morph had no effect, with males and hermaphrodites performing equivalently. This study is the first to unequivocally document the occurrence of a male function of hermaphrodites in a natural population of an androdioecious species.     

Androdioecy in the entomophilous tree Fraxinus ornus (Oleaceae)

Androdioecy (the coexistence of two genders, cosexuals and males, in a single population) is a rare breeding system. In terms of functional gamete production, androdioecy has been reported in a small number of wind-pollinated and insect-pollinated species. In this study we examine the floral biology, stability of gender, and fruit and seed production after self-pollination and outcrossing pollination in a potentially androdioecious tree, Fraxinus ornus , in southern France. Individual trees have either hermaphrodite flowers or male flowers, the latter lacking a well-developed gynoecium. The two genders produce morphologically similar pollen and were present in 1:1 or slightly male-biased ratios. Only hermaphrodites set fruit during 2–4 yr of observation at four different sites in southern France. Experimental pollinations of eight trees in two different sites showed that hermaphrodites produce viable pollen in dehiscent anthers and viable seeds. Thus, in terms of functional gamete production, F. ornus is a new case of functional androdioecy. Seven hermaphrodites were self-compatible, and such self-compatibility of hermaphrodites might improve the colonizing ability of F. ornus in a region where this species is actively expanding its range.     

The differentiation and development of pistils of hermaphrodites and pistillodes of males in androdioecious Osmanthus fragrans L. and implications for the evolution to androdioecy

The evolutionary pathway between hermaphroditism and dioecy draws widespread interests, and androdioecy is rarely achieved as an intermediate state between the two breeding systems. Flower bud differentiations in the pistils of hermaphrodites and the pistillodes of males in androdioecious Osmanthus fragrans L. were investigated by paraffin sectioning to elucidate the evolution to androdioecy. Results showed that the regularity and rhythm in flower bud differentiation between males and hermaphrodites were almost consistent and included six main stages. However, the hermaphrodites always lagged behind the males at each stage. The apical floret in the same inflorescence developed earlier than did the lateral ones in both hermaphrodites and males. The most significant difference between males and hermaphrodites was observed at the carpel differentiation stage. Two carpel primordia appeared inside the stamens of both males and hermaphrodites at the initial stage. These two carpels gradually fused with each other in hermaphrodites and eventually developed into a normal pistil with a stigma, a style, and an ovary. However, a cavity grew conspicuously over time between two carpels as developed in males. The two carpels eventually developed into a pistillode with two independent bracteal tissues. However, from the whole development process, the male retained the developmental residue of the hermaphrodite. Thus, the pistillodes of males could be traced to the pistils of hermaphrodites. This finding shows that males may be derived from hermaphrodites in O. fragrans. On the basis of this finding and previous studies on Oleaceae, androdioecy could be regarded as a transition from hermaphroditism to dioecy in this family.     

Germination of Excised Fraxinus ornus Embryos With and Without Phleomycin

In excised embryos of Fraxinus ornus, the first macroscopic sign of germination is a curvature near the root apex. Histological studies show that this curvature is due to cell elongation on the convex side and is accompanied by cell division. Metabolic changes, manifested by the disintegration of protein bodies and tissue differentiation, also occur during this time. In embryos treated with phleomycin cell division is completely inhibited but elongation, root hair formation, disintegration of protein bodies, and differentiation are still detectable. Phleomycin also drastically inhibits the formation of chlorophylls and the fresh weight increase of embryos.     

Lateral root emission in woody taproots of Fraxinus ornus L.

Abstract

We applied environmental stresses, namely dehydration, pruning and bending, to woody taproots of Fraxinus ornus L. in order to: (i) identify a method that could be applied in routine studies of lateral root development from a secondary structure; and (ii) carry out anatomical investigations to identify the tissue involved in the recruitment of lateral root mother cells (LRMC). We found that all three methods induce the formation of new lateral roots from a woody parental root. However, bending stress considerably reduced the zone of the woody parental root (the curvature) for analysis when studying the process of emission of a new lateral root. The trace left by a new lateral root in the taproot secondary xylem forms a V-shaped insertion zone that starts in contact with a growth ring and enlarges toward the periphery. This type of insertion zone suggests that the vascular cambium is the tissue-source of initials that produce the root primordium of a new lateral root. In the case of root bending, the emission of a new lateral root occurs also in the convex side of the curvature and is preceded by the formation, at the same site, of a small amount of reaction wood. Thus, reaction wood and lateral root emission are two aspects of the same response mechanism to bending. Consequently, anatomical and cytological studies of lateral root formation should focus on this part of the woody taproot. By peeling off the bark at this site, one has direct access to the underlying living tissues and can thus investigate lateral root formation also at a biochemical and molecular level.     

Communities of fungal endophytes in leaves of Fraxinus ornus are highly diverse

Communities of endophytic fungi in leaves of Manna ash (Fraxinus ornus) were examined to both the north and south of the Alps, i.e. within and beyond the native range of this tree species. Almost all leaves examined had been colonized by endophytic fungi. One hundred and two morphotypes were found, and 62 of them were identified to genus or species level using ITS sequencing and micromorphology. Venturia orni was most frequent and occurred in almost one third (32%) of the 1536 examined leaf segments. It was five times more abundant than Colletotrichum acutatum, the second most frequent endophyte. Other frequently isolated endophytes include Paraconiothyrium sp. 1, Mycosphaerella aurantia, Septoria cretae, Botryosphaeria dothidea and Boeremia exigua. The ash dieback pathogen was not isolated. The endophyte communities differed between the north and south of the Alps and the individual tree types had a distinct influence within sites.     

Similar gender dimorphism in the costs of reproduction across the geographic range of Fraxinus ornus

BACKGROUND AND AIMS: The reproductive costs for individuals with the female function have been hypothesized to be greater than for those with the male function because the allocation unit per female flower is very high due to the necessity to nurture the embryos until seed dispersal occurs, while the male reproductive allocation per flower is lower because it finishes once pollen is shed. Consequently, males may invest more resources in growth than females. This prediction was tested across a wide geographical range in a tree with a dimorphic breeding system (Fraxinus ornus) consisting of males and hermaphrodites functioning as females. The contrasting ecological conditions found across the geographical range allowed the evaluation of the hypothesis that the reproductive costs of sexual dimorphism varies with environmental stressors. METHODS: By using random-effects meta-analysis, the differences in the reproductive and vegetative investment of male and hermaphrodite trees of F. ornus were analysed in 10 populations from the northern (Slovakia), south-eastern (Greece) and south-western (Spain) limits of its European distribution. The variation in gender-dimorphism with environmental stress was analysed by running a meta-regression between these effect sizes and the two environmental stress indicators: one related to temperature (the frost-free period) and another related to water availability (moisture deficit). KEY RESULTS: Most of the effect sizes showed that males produced more flowers and grew more quickly than hermaphrodites. Gender differences in reproduction and growth were not minimized or maximized under adverse climatic conditions such as short frost-free periods or severe aridity. CONCLUSIONS: The lower costs of reproduction for F. ornus males allow them to grow more quickly than hermaphrodites, although such differences in sex-specific reproductive costs are not magnified under stressful conditions.     

Functional males in pair‐mating outcrossing hermaphrodites

In the mating system of simultaneously hermaphroditic animals, sexual allocation is predicted to vary as a function of the number of potential mates. According to the Hermaphrodite's Dilemma, sexual conflict over the preferred sexual role in hermaphroditic animals is resolved by reciprocity (i.e. by alternating sexual roles), accompanied by the animals' occasional cheating in the preferred role at a relatively low frequency. In a 350‐generation‐old laboratory strain of the pair‐mating outcrossing hermaphroditic polychaete worm Ophryotrocha diadema, we show that 9% of the individuals mated only in the male role over long periods, indicating a male‐role preference (temporary functional males). Furthermore, 2% of the individuals mated for their whole lifetime exclusively as males (permanent functional males). These findings indicate that the sex allocation of some individuals may vary from the predicted optimal sex allocation for the population. Morphologically, functional males exhibited a hermaphroditic phenotype (i.e. they matured a single batch of oocytes that they never laid and acted as functional males). We show that temporary functional males appeared in hermaphroditic populations under promiscuous mating regimes significantly more often than under monogamous ones. Indeed, under promiscuity, there are many mating opportunities and O. diadema hermaphrodites compete for mates, whereas, under monogamy, the two partners regularly take turns in laying cocoons and fertilizing their partner's cocoon. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100 , 451–456.     

Accidental X-Y recombination and the aetiology of XX males and true hermaphrodites

Accidental recombination between the differential segments of the X and Y chromosomes in man occasionally allows transfer of Y-linked sequences to the X chromosome leading to testis differentiation in so-called XX males. Loss of the same sequences by X-Y interchange allows female differentiation in a small proportion of individuals with XY gonadal dysgenesis. A candidate gene responsible for primary sex determination has recently been cloned from within this part of the Y chromosome by Page and his colleagues. The observation that a homologue of this gene is present on the short arm of the X chromosome and is subject to X-inactivation, raises the intriguing possibility that sex determination in man is a quantitative trait. Males have two active doses of the gonad determining gene, and females have one dose. This hypothesis has been tested in a series of XX males, XY females and XX true hermaphrodites by using a genomic probe, CMPXY1, obtained by probing a Y-specific DNA library with synthetic oligonucleotides based on the predicted amino-acid sequence of the sex-determining protein. The findings in most cases are consistent with the hypothesis of homologous gonad-determining genes, GDX and GDY, carried by the X and Y chromosomes respectively. It is postulated that in sporadic or familial XX true hermaphrodites one of the GDX loci escapes X-inactivation because of mutation or chromosomal rearrangement, resulting in mosaicism for testis and ovary-determining cell lines in somatic cells. Y-negative XX males belong to the same clinical spectrum as XX true hermaphrodites, and gonadal dysgenesis in some XY females may be due to sporadic or familial mutations of GDX.     

Visible foliar injury and physiological responses to ozone in Italian provenances of Fraxinus excelsior and F. ornus

We compared leaf visible injury and physiological responses (gas exchange and chlorophyll a fluorescence) to high O3 exposure (150 nmol mol(-1) h, 8 h day(-1), 35-40 days) of two woody species of the same genus with different ecological features: the mesophilic green ash (Fraxinus excelsior) and the xerotolerant manna ash (F. ornus). We also studied how provenances from northern (Piedmont) and central (Tuscany) Italy, within the two species, responded to O3 exposure. Onset and extent of visible foliar injury suggested that F. excelsior was more O3 sensitive than F. ornus. The higher stomatal conductance in F. ornus than in F. excelsior suggested a larger potential O3 uptake, in disagreement to lower visible foliar injury. The higher carbon assimilation in F. ornus suggested a higher potential of O3 detoxification and/or repair. Contrasting geographical variations of ash sensitivity to O3 were recorded, as Piedmont provenances reduced gas exchange less than Tuscan provenances in F. excelsior and more in F. ornus. Visible injury was earlier and more severe in F. excelsior from Piedmont than from Tuscany, while the provenance did not affect visible injury onset and extent in F. ornus.     

The evolution from females to hermaphrodites results in a sexual conflict over mating in androdioecious nematode worms and clam shrimp

The nematode worm Caenorhabditis elegans and the clam shrimp Eulimnadia texana are two well‐studied androdioecious species consisting mostly of self‐fertilizing hermaphrodites and few males. To understand how androdioecy can evolve, a simple two‐step mathematical model of the evolutionary pathway from a male–female species to a selfing‐hermaphrodite species is constructed. First, the frequency of mutant females capable of facultative self‐fertilization increases if the benefits of reproductive assurance exceed the cost. Second, hermaphrodites become obligate self‐fertilizers if the fitness of selfed offspring exceeds one‐half the fitness of outcrossed offspring. Genetic considerations specific to C. elegans and E. texana show that males may endure as descendants of the ancestral male–female species. These models combined with an extensive literature review suggest a sexual conflict over mating in these androdioecious species: selection favours hermaphrodites that self and males that outcross. The strength of selection on hermaphrodites and males differs, however. Males that fail to outcross suffer a genetic death. Hermaphrodites may never encounter a rare male, and those that do and outcross only bear less fecund offspring. This asymmetric sexual conflict results in an evolutionary stand‐off: rare, but persistent males occasionally fertilize common, but reluctant hermaphrodites. A consequence of this stand‐off may be an increase in the longevity of the androdioecious mating system.     

When males and hermaphrodites coexist: a review of androdioecy in animals

Androdioecy (populations consisting of males and hermaphrodites)is a rare mating system in plants and animals: up to 50 plantsand only 36 animals have been described as being androdioecious,with most of the latter being crustaceans. To date, a thoroughcomparative analysis of androdioecy in animals has not beenundertaken. Herein we present such an analysis. Androdioecyhas only been extensively surveyed in 2 animal taxa: the nematodeCaenorhabditis and the clam shrimp Eulimnadia. The other majortaxon having androdioecious species is the Cirripedia (barnacles),but there are only limited studies on androdioecy in this group.In animals, androdioecy is found either in species that havemorphologically and ecologically distinct sexes (that is, hermaphroditesand small, "complemental" males) that are derived from hermaphroditicancestors (that is, the barnacles) or in species that have similarly-sizedmales and hermaphrodites that have been derived from dioeciousancestors (the remaining androdioecious species). We suggestthat the barnacles have evolved a sexual specialization in theform of these complemental males that can more efficiently usethe constrained habitats that these barnacles often experience.For the remaining species, we suggest that androdioecy has evolvedas a response to reproductive assurance in species that experienceepisodic low densities. Additionally, we hypothesize that thedevelopment of mechanisms allowing reproductive assurance inspecies with a number of sexually differentiated traits is mostlikely to result in androdioecy rather than gynodioecy (mixturesof females and hermaphrodites), and that these species may bedevelopmentally constrained to stay androdioecious rather thanbeing capable of evolving into populations solely consistingof efficient, self-compatible hermaphrodites. We conclude bysuggesting several areas in need of further study to understandmore completely the evolution and distribution of this interestingmating system in animals.     

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.     

Density-dependent self-fertilization and male versus hermaphrodite siring success in an androdioecious plant

Models of mating-system evolution emphasize the importance of frequency-dependent interactions among mating partners. It is also known that outcross siring success and the selfing rate in self-compatible hermaphrodites can be density dependent. Here, we use array experiments to show that the mating system (i.e., the outcrossing rate) and the siring success of morphs with divergent sex allocation strategies are both density dependent and frequency dependent in androdioecious populations of the wind-pollinated, annual plant Mercurialis annua. In particular, the outcrossing rate is a decreasing function of the mean interplant distance, regulated by a negative exponential pollen fall-off curve. Our results indicate that pollen dispersed from a male inflorescence are over 60% more likely to sire outcrossed progeny than equivalent pollen dispersed from hermaphrodites, likely due to the fact that males, but not hermaphrodites, disperse their pollen from erect inflorescence stalks. Because of this difference, and because males of M. annua produce much more pollen than hermaphrodites, the presence of males in the experimental arrays reduced both the selfing rate and the outcross siring success of hermaphrodites. We use our results to infer a density threshold below which males are unable to persist with hermaphrodites but above which they can invade hermaphroditic populations. We discuss our findings in the context of a metapopulation model, in which males can only persist in well-established populations but are excluded from small, sparse populations, for example, in the early stages of colonization.     

植物生活史繁殖对策与干扰关系的研究

植物生活史繁殖对策研究是涉及植物的适应或进化、生态系统退化与恢复过程、生物多样性保护等多方面理论生态学和应用生态学研究内容的生态学研究领域。按Grime的植物生活史繁殖对策分类、植物营养繁殖与干扰适应、种子形态学与干扰适应、土壤种子库与干扰适应、植物繁殖体传播和萌发与干扰适应论述了当今极受关注的植物生活史繁殖对策与干扰关系,简述了我国干旱区干扰与植物生活史繁殖对策关系研究。     

Males outcompete hermaphrodites for seed siring success in controlled crosses in the polygamous Fraxinus excelsior (Oleaceae)

Polygamy (including trioecy and subdioecy), the co-occurrence of males, hermaphrodites, and females in natural populations, is a rare and poorly studied breeding system expressed in Fraxinus excelsior L. (Oleaceae), a wind-pollinated tree. Here we investigate siring ability of pollen from male vs. hermaphrodite individuals to better understand this sex polymorphism. We conducted single-donor and two-donor pollination experiments and compared both fruit set and seed siring success, assessed with polymorphic microsatellite markers, of male and hermaphrodite individuals. Single pollen donor crosses allowed us to verify the male function of hermaphrodites. However, pollen from hermaphrodites was much less proficient than male pollen, with males siring 10 times as many fruits in single donor pollination treatments. This result was strengthened by the surprisingly low reproductive success of hermaphrodites in pollen competition conditions: of the 110 seedlings analyzed three were selfed and only one was sired by the hermaphrodite donor. The remaining 106 were sired by the male pollen donor. These results raise the question of the maintenance of male fertility in hermaphrodites in Fraxinus excelsior. Male function of hermaphrodites in this species now needs to be assessed under field conditions.     

Selection in plant populations of effectively infinite size: IV. The maintenance of males among hermaphrodites for a biallelic model

Conditions for the maintenance of males in androdioecious populations (populations with both male and hermaphrodite individuals) have been derived for four different one-locus two-allele models of inheritance of androdioecy. The results are not in general accordance with those already known: depending on the mode of inheritance, males can be maintained irrespective of their fertilities. If males are sufficiently fertile, it may happen that they are maintained only for intermediate selfing rates of the hermaphrodites. A result already found for gynodioecy is confirmed for androdioecy, namely, that a 11 sex ratio is immediately established among zygotes if hermaphrodites appear as heterozygotes only.     

Genetic analysis of 38XX males with genital ambiguities and true hermaphrodites in pigs

In pig, the frequency of intersexuality ranges from 0.1 to O.6%, depending on the breed. In a closed pig herd at INRA an intersex condition was observed in 0.75% of ‘females’. The present study describes 11 animals with a 38XX karyotype and the presence of testicular tissue. Phenotypically, all presented with abnormal external or/and internal genitalia. Southern blot analysis with Y-specific probes (SRY and ZFY) revealed the absence of Y material in all animals tested. By polymerase chain reaction (PCR) amplification, 10 of 11 intersex pigs lacked the SRY gene in gonad DNA. These data are compatible with an autosomally (or pseudoautosomally) determined mechanism. Moreover, analysis of familial cases seemed to indicate that 38XX male pseudohermaprodites and 38XX true hermaphrodites may represent alternative manifestations of the same genetic defect.     

Allee effect and self-fertilization in hermaphrodites: reproductive assurance in a structured metapopulation

Reproductive assurance through selfing during colonization events or when population densities are low has often been put forward as a mechanism selecting for the evolution of self-fertilization. Such arguments emphasize on the role of both local demography and metapopulation processes. We developed a model for the evolution of self-fertilization in a structured metapopulation in which local densities are not steady because of population growth. Reproduction by selfing is density-independent (reproductive assurance) but selfed seeds endure inbreeding depression, whereas reproduction by outcrossing is density-dependent (Allee effect). First, we derived an analytical criterion for metapopulation viability as a function of the selfing rate and metapopulation parameters. We show that outcrossers can develop a viable metapopulation when they produce a high amount of dispersal seeds that counterbalances their incapacity to found new populations from low densities. Second, the model shows there is a positive feedback between demography and outcrossing rates, leading to either complete outcrossing or selfing. Specifically, we illustrate that inbreeding depression can paradoxically favor the evolution of selfing because of its negative effect on density. Also, complete outcrossing can be selected despite pollen limitation, although it does not provide a full seed set. This model underlines the influence of the mating system both on demography and gene dynamics in a metapopulation context.