Spatial structure of nuclear factors involved in sex determination in the gynodioecious Thymus vulgaris L.

Theoretical models have shown that the maintenance of high frequencies of females in gynodioecious species can be explained by the spatial structure of cytoplasmic and nuclear genes involved in sex determination. Whereas spatial structure of cytoplasmic factors which cause male-sterility has been studied in several taxa, that of nuclear factors that restore male-fertility (restoration factors) has received little attention. In this paper, we estimate spatial variation in the frequency of restoration factors associated with different cytoplasmic male-sterilities in the gynodioecious Thymus vulgaris. Clonal replicates of five female plants bearing at least four different male-sterilities and originating from five different populations (i.e., five cytogenotypes), were reciprocally transplanted into the original populations. Following open pollination at each site, seeds were harvested, germinated and grown to flowering. The frequency of hermaphrodites in each progeny was used to estimate the frequency of restoration factors in each population. For all cytogenotypes, there was marked variation in the rate of restoration among populations, indicating that spatial structure in nuclear restoration factors may influence the variation in female frequency in T. vulgaris. For three out of the five cytogenotypes, higher restoration rates were obtained for females introduced into their original population, which agrees with the theoretical prediction, under some hypotheses, that restoration factors are selected for in populations that contain the associated male-sterility (Gouyon and Couvet, 1985). The very low restoration rate in the progeny of one female clone in three populations indicates that stochastic events, such as founder effects, could lead to a local absence of restoration factors and thus the high female frequency (up to 90%) observed in some populations. Differences among cytogenotypes suggest that some male-sterilities are associated with rarer restoration factors and are thus more likely to cause patches of females to develop in young populations.     

Effective population size and tests of neutrality at cytoplasmic genes in Arabidopsis.

Cytoplasmic genomes typically lack recombination, implying that genetic hitch-hiking could be a predominant force structuring nucleotide polymorphism in the chloroplast and mitochondria. We test this hypothesis by analysing nucleotide polymorphism data at 28 loci across the chloroplast and mitochondria of the outcrossing plant Arabidopsis lyrata, and compare patterns with multiple nuclear loci, and the highly selfing Arabidopsis thaliana. The maximum likelihood estimate of the ratio of effective population size at cytoplasmic relative to nuclear genes in A. lyrata does not depart from the neutral expectation of 0.5. Similarly, the ratio of effective size in A. thaliana is close to unity, the neutral expectation for a highly selfing species. The results are thus consistent with neutral organelle polymorphism in these species or with comparable effects of hitch-hiking in both cytoplasmic and nuclear genes, in contrast to the results of recent studies on gynodioecious taxa. The four-gamete test and composite likelihood estimation provide evidence for very low levels of recombination in the organelles of A. lyrata, although permutation tests do not suggest that adjacent polymorphic sites are more closely linked than more distant sites across the two genomes, suggesting that mutation hotspots or very low rates of gene conversion could explain the data.     

Testing for sex differences in biparental inbreeding and its consequences in a gynodioecious species

In many gynodioecious species cross-pollinated seeds from females outperform those from hermaphrodites. Using the gynodioecious alpine perennial Silene acaulis, I investigated whether this was the result of greater biparental inbreeding among hermaphrodites leading to greater biparental inbreeding depression. I also determined the influence of relatedness on progeny fitness. Experiments were performed using individuals from a site whose population structure and coefficient of inbreeding was known. In the first experiment, crosses were made on plants in the field to determine the effect of seven different crossing distances, plus selfing, on germination and early seedling survival and growth. Although selfed seeds died more often and grew slower than crossed seeds, the effect of crossing distance was negligible for all measured fitness traits, refuting the biparental inbreeding hypothesis as a mechanism to explain why seeds from hermaphrodites die more often than those from females. Nonetheless, cross-pollinated seeds from hermaphrodites did die more, indicating that another mechanism must be responsible. In the second experiment, the effect of different levels of inbreeding on germination and seedling survival was determined by growing seeds from experimental matings varying in relatedness. Inbreeding depression for a multiplicative fitness estimate was significant for all levels of inbreeding, suggesting that inbred individuals are unlikely to become established in the population and providing insight into the results of the first experiment. Alternative hypotheses are discussed to explain why seeds from hermaphrodites die more often, which together with the results of this study, suggest that the restoration of male function in hermaphrodites comes with a correlated cost to seedling survival.     

The role of the Y-chromosome in sex determination.

The basic plan of gonadal development in both sexes is female unless testes are induced by factor(s) of the Y chromosome, known as testis determining factor(s) (TDF). It is not clearly established whether the Y chromosome control is autonomous or under the control of a gene on the X chromosome or autosomes. A gene for the H-Y antigen (Histocompatibility-Y antigen) has been postulated to be the factor determining testicular differentiation. Recent studies have demonstrated that the gene for testis determination and the H-Y determinant are two separate entities. Although earlier cytogenetic observations localized TDF on the pericentric region of the short arm of the Y chromosome, subsequent findings by high-resolution chromosome banding and molecular analysis localise TDF to the distal part of the short arm of the Y chromosome, adjacent to the pseudoautosomal region. A candidate for TDF, the ZFY, was localised within the 140 kb interval where the position of TDF was defined, and considered as the TDF gene. However, a smaller gene sequence of 35 kb, the SRY, situated outside the 140 kb ZFY region, has recently been isolated and proved to be the only and the smallest part of the Y chromosome necessary for male sex determination.     

The nearness of you: the effect of population structure on siring success in a gynodioecious species

Theoretically, both balancing selection and genetic drift can contribute to the maintenance of gender polymorphism within and/or among populations. However, if strong differences exist among genotypes in the quantity of viable gametes they produce, then it is expected that these differences will play an important role in determining the relative frequency of the genotypes and contribute to whether or not such polymorphism is maintained. In this issue, De Cauwer et al. (2010) describe an investigation of gynodioecious wild sea beet, which in addition to containing females, contain two types of hermaphrodites: restored hermaphrodites carrying a cytoplasm that causes pollen sterility and a nuclear gene that restores pollen fertility, and hermaphrodites without the sterilizing cytoplasm. The results show that restored hermaphrodites, who have relatively low pollen viability, achieve disproportionately high siring success simply because of where they are located in a patchy population ( Fig. 1 ). Notably, these individuals tend to be close to females because of the genetics of sex determination. These results indicate that population structure caused by drift processes can have an unexpectedly large effect on the fitness of these low quality hermaphrodites, thereby contributing in the short term to the maintenance of gynodioecy in this population. While these results indicate that population structure caused by drift processes can have a large effect on the relative fitness of genetic variants, whether these effects promote or discourage the maintenance of polymorphism in the long term is still up for debate.

Figure 1 Open in figure viewer PowerPoint A stretch of beach along which wild sea beet can be seen to be growing among the rocks above the splash zone. This linear arrangement enhances the potential for mating success to depend on proximity to other plants (Photo: J.‐F. Arnaud).     

Identifying genes for male sex determination in humans.

The convergence of genetic and molecular technologies has led to the identification of a number of genes for male sex determination. The observation of chromosomal translocations, deletions, and duplications in sex reversed individuals was instrumental for the positional cloning of SRY, SOX9, WT1, and DAX1. Cloning by protein-DNA interaction was required for the identification of SF1. The observation of an extended phenotype for the alpha thalassemia-mental retardation syndrome assigned a role for XH2 in the testicular determining process. Over the next several years, new sex determining genes will be identified by linkage analysis in large families with multiple sex reversed members, comparative genomic hybridization of sex reversed individuals, and database searches for genes that encode interacting proteins or paralogs of other species. Given the apparent differences in the sex determining mechanisms of even closely related species, the roles of all of these genes will require confirmation by demonstrating expression in human gonadal ridge at the critical time, and that mutations result in sex reversal.     

Modelling the maintenance of male-fertile cytoplasm in a gynodioecious population

Gynodioecy is the co-occurrence of females and hermaphrodites in populations. It is usually due to the combined action of cytoplasmic male sterility (CMS) genes and nuclear genes that restore male fertility. According to previous theoretical studies, it is very difficult to explain the maintenance of gynodioecy with CMS and male-fertile cytotypes, although it has been observed in some species. However, only very specific situations have been investigated so far. We present a model to investigate the conditions that promote the maintenance of this breeding system in the case of an outcrossed species when CMS and male-fertile (non-CMS) cytotypes are present in an infinite panmictic population. We show that the type of cost of restoration strongly affects the conditions for stable maintenance of gynodioecy. Stable nuclear-cytoplasmic gynodioecy requires a female advantage, which is a classical condition for gynodioecy, but also a cost of CMS for female fitness, which had been rarely investigated. A cost of restoration is also needed, which could affect either pollen or seeds. Finally, we found that gynodioecy was attainable for a large set of parameter values, including low differences in fitness among genotypes and phenotypes. Our theoretical predictions are compared with previous theoretical work and with results of empirical studies on various gynodioecious species.     

Founder effects and sex ratio in the gynodioecious Thymus vulgaris L.

Thymus vulgaris is a gynodioecious species (in which females and hermaphrodites coexist) with a highly variable frequency of females among natural populations (5–95%) and a high average female frequency (60%). Sex determination involves both cytoplasmic genes responsible for male sterility, i.e. the female phenotype, and specific nuclear factors responsible for the restoration of male fertility, and thus a hermaphrodite phenotype. In this study, molecular markers of the mitochondrial genome have been used to quantify the cytoplasmic diversity in 11 clumps of individuals observed in four recently founded populations. The very low diversity within patches in conjunction with the strong diversity among patches strongly suggests that clumps of individuals are the result of single matrilinear families. In clumps that contain mainly females, all the analysed females showed the same cytoplasmic pattern. This pattern differed from that shown by neighbouring hermaphrodites, indicating that the determination of sex is locally cytoplasmic. A comparison of genetic diversity before and after fire in one population showed that disturbances may cause a reduction in genetic diversity and a concurrent induction of local cytoplasmic determination of sex. Such cytoplasmic determination of sex in colonizing populations, together with the greater seed set of females, may largely improve the colonizing ability of the species.     

The biochemical role of SRY in sex determination

The human sex-determining gene on the Y chromosome, termed SRY, has recently been isolated by positional cloning; compelling evidence now exists equating SRY with the testis-determing factor, TDF. The SRY gene product is an HMG box protein whose DNA-binding activity is vital for testis formation as sex-reversed patients with SRY mutations lack this activity in vitro. The in vivo DNA target for SRY, however, remains elusive. Here, we show, by gel retardation analysis, that SRY recognises specific DNA sequences and that such sequences exist upstream of the AMH promoter, a potential downstream target for SRY. We also describe the DNA bending and cruciform DNA-binding functions of SRY and propose a model for the potential action of SRY in the “HMG-1-rich” mammalian nucleus. © 1994 Wiley-Liss, Inc.     

The origin and characterization of new nuclear genes originating from a cytoplasmic organellar genome

Endosymbiotic transfer of DNA and functional genes from the cytoplasmic organelles (mitochondria and chloroplasts) to the nucleus has been a major factor driving the origin of new nuclear genes, a process central to eukaryote evolution. Although organelle DNA transfers very frequently to the nucleus, most is quickly deleted, decays, or is alternatively scrapped. However, a very small proportion of it gives rise, immediately or eventually, to functional genes. To simulate the process of functional transfer, we screened for nuclear activation of a chloroplast reporter gene aadA, which had been transferred from the chloroplast to independent nuclear loci in 16 different plant lines. Cryptic nuclear activity of the chloroplast promoter was revealed, which became conspicuous when present in multiple nuclear copies. We screened ～50 million cells of each line and retrieved three plants in which aadA showed strong nuclear activation. Activation occurred by acquisition of the CaMV 35S nuclear promoter or by nuclear activation of the native chloroplast promoter. Two fortuitous sites within the 3' UTR of aadA mRNA both promoted polyadenylation without any sequence change. Complete characterization of one nuclear sequence before and after gene transfer demonstrated integration by nonhomologous end joining involving simultaneous insertion of multiple chloroplast DNA fragments. The real-time observation of three different means by which a chloroplast gene can become expressed in the nucleus suggests that the process, though rare, may be more readily achieved than previously envisaged.     

The transmission of genes via pollen and ovules in gynodioecious angiosperms

Gynodioecy is defined as the class of sexual dimorphism in which females are constant (functionally unisexual) and males are inconstant (bisexual). Quantitative measures describe the sexual performance of males in gynodioecious populations. The relative (male/female) ovule contribution compares the ovule contributions of the sexes to the next generation. Male constancy and inconstancy, the proportions of male genes transmitted via pollen and ovules, are direct measure of maleness and femaleness. Equations permitting the values of the sex parameters to be estimated are derived. The values obtained for twelve species show that gynodioecy is not a single breeding system, but merges into hermaphroditism and dioecy at opposite extremes. The distribution of inconstancies among individual males is described for six species. In most species, seed counts overestimate the ovule contributions of males.     

Female frequencies and fitness components between sex phenotypes among gynodioecious populations of the colonizing species Trifolium hirtum All. in California

Summary Male sterility has been recently discovered in Californian populations of rose clover (Trifolium hirtum). This study describes the frequency distribution of male sterility in Turkish and Californian populations, and compares fitness components of hermaphrodites and females. As male-steriles were found in Turkey, it is likely that they were introduced to California during the 1940's with the original material derived from Turkey. The spread of male-sterile genotypes in California has given rise to an asymmetrical frequency distribution of male sterility with positive skewness. The frequency of females has not exceeded fifty percent and it appears to be temporally stable in most of the Californian populations. The hypothesis that female frequencies and fitness differences between phenotypes are correlated was tested by comparing sex phenotypes in seven populations with contrasting levels of male sterility. The analysis of those populations showed no evidence for such a correlation as no significant differences were found between sex phenotypes for fecundity and seed germination. The hypothesis that females are maintained due to fitness differences in the progeny of hermaphrodites and females was experimentally tested in the population with maximum frequency of male-steriles. The results showed no significant differences in the demographic performance of the progenies of hermaphrodites and females. The present results are discussed in terms of the possible mechanism of maintenance of gynodioecy in rose clover.     

Consequences of prairie fragmentation on the progeny sex ratio of a gynodioecious species, Lobelia spicata (Campanulaceae)

Habitat fragmentation of prairie ecosystems has resulted in increased isolation and decreased size of plant populations. In large populations, frequency-dependent selection is expected to maintain genetic diversity of sex determining factors associated with gynodioecy, that is, nuclear restorer genes that reverse cytoplasmic male sterility (nucleocytoplasmic gynodioecy). However, genetic drift will have a greater influence on small isolated populations that result from habitat fragmentation. The genetic model for nucleocytoplasmic gynodioecy implies that the proportion of female progeny produced by hermaphroditic and female plants will show more extreme differences in populations with reduced allelic diversity, and that restoration of male function will increase with inbreeding. We investigated potential impacts of effects resulting from reduced population sizes by comparison of progeny sex ratios produced by female and hermaphroditic plants in small and large populations of the gynodioecious prairie species, Lobelia spicata. A four-way contingency analysis of the impact of population size, population sex ratio, and maternal gender on progeny sex ratios showed that progeny sex ratios of hermaphroditic plants were strongly influenced by population size, whereas progeny sex ratios of female plants were strongly influenced by population sex ratio. Further, analysis of variation in progeny-type distribution indicated decreased restoration and increased loss of male function in smaller and isolated populations. These results are consistent with reduced allelic diversity or low allelic frequency at restorer loci in small and isolated populations. The consequent decrease in male function has the potential to impede seed production in these fragmented prairies.     

Coexistence of cytoplasmic and nuclear genes for male sterility in Petunia

Summary Cytoplasmic male sterility (cms) and nuclear male sterility (nms) in Petunia were described respectively as possible autonomous and integrated states of the same genetic element by Frankel (1971). In the present study we describe genetic analysis of the interaction between the cms, the nuclear gene for male sterility (e) and the fertility restorer allele (Rf). The main findings in this study are: (1) The nuclear sterility allele can coexist in one or two dosages with the cytoplasmic male sterility elements (ste) in somatic cells or female gametes; (2) the presence of the fertility restorer allele Rf is not required for the coexistence of ste and e and (3) Rf does not interact epistatically with e, e.g., the expression of e is independent of Rf—the genotypes (S) RfRfee and (S) Rfrfee are male sterile.Contribution from the Agricultural Research Organization. The Volcani Center, Bet Dagan, Israel. 1983 series No. 846 E     

Mating system, sex ratio, and persistence of females in the gynodioecious shrub Daphne laureola L. (Thymelaeaceae)

Although in gynodioecious populations male steriles require a fecundity advantage to compensate for their gametic disadvantage, southern Spanish populations of the long-lived shrub Daphne laureola do not show any fecundity advantage over hermaphrodites in terms of seed production and early seedling establishment. By using allozyme markers, we assess the mating system of this species in five populations differing in sex ratio, and infer levels of inbreeding depression over the whole life cycle by comparing the inbreeding coefficients at the seed and adult plant stages. Extremely low outcrossing rates (0.001相似文献   

Observation of a ZZW female in a natural population: implications for avian sex determination

Avian sex determination is chromosomal; however, the underlying mechanisms are not yet understood. There is no conclusive evidence for either of two proposed mechanisms: a dominant genetic switch or a dosage mechanism. No dominant sex-determining gene on the female-specific W chromosome has been found. Birds lack inactivation of one of the Z chromosomes in males, but seem to compensate for a double dose of Z-linked genes by other mechanisms. Recent studies showing female-specific expression of two genes may support an active role of the W chromosome. To resolve the question of avian sex determination the investigation of birds with a 2A: ZZW or 2A: ZO genotype would be decisive. Here, we report the case of an apparent 2A: ZZW great reed warbler (Acrocephalus arundinaceus) female breeding in a natural population, which was detected using Z-linked microsatellites. Our data strongly suggest a role of W-linked genes in avian sex determination.