高等植物性别分化研究进展

高等植物性别分化研究主要包括三个方面：性别分化特异大分子标记物的鉴定(分化程序)；诱导信号(如植物激素)的分析和性决定基因的分离与分析。近年来，植物性别分化研究取得了较大进展，本文主要介绍这一研究在分化程序、诱导信号和性决定基因等方面的研究进展。     

Evolution of the pathways of sex differentiation in plants

The stages of formulating the theory of sex evolution, determination, and expression in plants were briefly reviewed. The role of phytohormones in sex differentiation in dioecious and monoecious plants was analyzed.     

GENETIC AND HORMONAL REGULATION OF GROWTH,FLOWERING, AND SEX EXPRESSION IN PLANTS

The interaction of the genetic and hormonal regulation of growth, flowering, and sex expression in plants is discussed. The genetic control of these processes is characterized, and data on their hormonal regulation are supplied. The interaction of genetic and hormonal regulation is considered with reference to tall-growing and genetic dwarf forms of the pea and wheat plants. It is shown that in the dwarf forms of the pea plant and in many other varieties, growth stimulation in response to treatment with the phytohormone gibberellic acid is clearly manifested and the expression of genetic dwarfism is eliminated, whereas in dwarf wheats it is expressed only slightly, if at all. At the same time both tall-growing and dwarf forms of both pea and wheat show a clearly defined growth retardation response to treatment with the growth inhibitor, abscisic acid, which causes the expression of physiological dwarfism. The short- and long-day characteristics of the photoperiodic response of plants are described as genetically controlled features, and data are given on the induction of flowering of a long-day variety coneflower grown under short-day conditions with the aid of gibberellins extracted from leaves of long-day vegetative plants of short-day Mammoth tobacco. Data are also supplied on the induction of flowering of a short-day variety, red-leaved goosefoot, grown under continuous light with the aid of metabolites extracted from leaves of the same Mammoth tobacco plants flowering under short-day conditions. This demonstrates the possibility of hormonal regulation of the genetically controlled long-day and short-day characteristics in photoperiodically sensitive plants. Genetic and hormonal regulation of sex expression in two dioecious plants, hemp and spinach, is discussed. It is shown that sex expression in these plants is regulated by gibberellins which are synthesized in leaves and cause male sex expression and by cytokinins which are synthesized in the roots and cause female sex expression. These data indicate that sex expression in dioecious plants is the result of interaction between the genetic apparatus and phytohormones.     

Monoecious Plants of Myriophyllum ussuriense (Regel) Maxim. in Japan

Myriophyllum ussuriense has been described as dioecious but monoecious plants were newly found from some populations in south-western Japan. Sex expression of monoecious plants proved labile and they sometimes bore male or female flowers alone. On the other hand, sex expression of dioecious plants was stable and seemed to be fixed genetically. M. ussuriense may be still in the course of differentiation from monoecy to dioecy. Received 13 April 2001/ Accepted in revised form 22 May 2001     

Role of Phytohormones in Sex Differentiation in Plants

The role of phytohormones in sex expression in plants is briefly surveyed. The interaction of hereditary and environmental factors in sex expression is considered. A major role of gibberellins and cytokinins in the regulation of sex expression in dioecious and monoecious flowering plants and in some muscoids is demonstrated. The evidence for the effect of other phytohormones and physiologically active compounds on sex expression in plants is examined.     

Slow evolution of sex‐biased genes in the reproductive tissue of the dioecious plant Salix viminalis

The relative rate of evolution for sex‐biased genes has often been used as a measure of the strength of sex‐specific selection. In contrast to studies in a wide variety of animals, far less is known about the molecular evolution of sex‐biased genes in plants, particularly in dioecious angiosperms. Here, we investigate the gene expression patterns and evolution of sex‐biased genes in the dioecious plant Salix viminalis. We observe lower rates of sequence evolution for male‐biased genes expressed in the reproductive tissue compared to unbiased and female‐biased genes. These results could be partially explained by the lower codon usage bias for male‐biased genes leading to elevated rates of synonymous substitutions compared to unbiased genes. However, the stronger haploid selection in the reproductive tissue of plants, together with pollen competition, would also lead to higher levels of purifying selection acting to remove deleterious variation. Future work should focus on the differential evolution of haploid‐ and diploid‐specific genes to understand the selective dynamics acting on these loci.     

Sex-Biased Temporal Gene Expression in Male and Female Floral Buds of Seabuckthorn (Hippophae rhamnoides)

Seabuckthorn is an economically important dioecious plant in which mechanism of sex determination is unknown. The study was conducted to identify seabuckthorn homologous genes involved in floral development which may have role in sex determination. Forty four putative Genes involved in sex determination (GISD) reported in model plants were shortlisted from literature survey, and twenty nine seabuckthorn homologous sequences were identified from available seabuckthorn genomic resources. Of these, 21 genes were found to differentially express in either male or female flower bud stages. HrCRY2 was significantly expressed in female flower buds only while HrCO had significant expression in male flowers only. Among the three male and female floral development stages (FDS), male stage II had significant expression of most of the GISD. Information on these sex-specific expressed genes will help in elucidating sex determination mechanism in seabuckthorn.     

Complex sex determination in the stinging nettle Urtica dioica

The dioecious species Urtica dioica harbours wide variation in sex ratio of seeds. We conducted a series of crosses to analyse the genetic basis of sex determination in this species. Dutch populations of U. dioica contain low proportions of monoecious individuals beside male and female plants. Self-pollination of monoecious plants always yielded female, male and monoecious plants, generally in a ratio of one female to three male/monoecious individuals. This motivated us to write down a simple model in which gender is determined by one major sex-determination locus with four alleles. In the model males and monoecious plants have distinct genotypes but are both heterozygous at the sex-determination locus. We first made crosses among progeny obtained after self-pollination of monoecious plants. These crosses showed that the monoecious trait generally showed Mendelian inheritance and was passed on to the next generation via both pollen and seeds. Further crosses between monoecious plants and plants from dioecious system indicated that alleles from the dioecious system are often dominant. However, many exceptions to our genetic model are observed which suggest that dominance is incomplete and/or that more genes are involved in sex determination. We discuss to what extent sex determination genes explain the strongly biased seed sex ratios and argue that additional genes, for instance genes for female choice, must also be involved.     

Sex‐specific floral morphology,biomass, and phytohormones associated with altitude in dioecious Populus cathayana populations

Relationships between sex‐specific floral traits and endogenous phytohormones associated with altitude are unknown particularly in dioecious trees. We thus examined the relationships between floral morphology or biomass and phytohormones in male and female flowers of dioecious Populus cathayana populations along an altitudinal gradient (1,500, 1,600, and 1,700 m above sea level) in the Xiaowutai Nature Reserve in northern China. The female and male flowers had the most stigma and pollen at 1,700 m, the largest ovaries and least pollen at 1,500 m, and the smallest ovaries and greater numbers of anthers at 1,600 m altitude. The single‐flower biomass was significantly greater in males than in females at 1,600 or 1,700 m, but the opposite was true at 1,500 m altitude. The biomass percentages were significantly higher in anthers than in stigmas at each altitude, while significantly greater gibberellin A3 (GA₃), zeatin riboside (ZR), indoleacetic acid (IAA), and abscisic acid (ABA) concentrations were found in female than in male flowers. Moreover, most flower morphological traits positively correlated with IAA in females but not in males. The biomass of a single flower was significantly positively correlated with ABA or IAA in males but negatively with ZR in females and was not correlated with GA₃ in both females and males. Our results demonstrate a distinct sexual adaptation between male and female flowers and that phytohormones are closely related to the size, shape, and biomass allocation in the pollination or fertilization organs of dioecious plants, although with variations in altitude.     

Pollination intensity influences sex ratios in dioecious Rumex nivalis, a wind-pollinated plant

Determining the mechanisms governing sex-ratio variation in dioecious organisms represents a central problem in evolutionary biology. It has been proposed that in plants with sex chromosomes competition between pollen tubes of female- versus male-determining microgametophytes (certation) causes female-biased primary sex ratios. Experimental support for this hypothesis is limited and recent workers have cast doubt on whether pollen-tube competition can modify sex ratios in dioecious plants. Here we investigate the influence of variation in pollination intensity on sex ratios in Rumex nivalis, a wind-pollinated alpine herb with strongly female-biased sex ratios. In a garden experiment, we experimentally manipulated pollination intensity using three concentric rings of female recipient plants at different distances from a central group of male pollen donors. This design enabled us to test the hypothesis that increasing pollen load size, by intensifying gametophyte competition, promotes female-biased sex ratios in R. nivalis. We detected a significant decline in pollen load at successive distance classes with concomitant reductions in seed set. Sex ratios of progeny were always female biased, but plants at the closest distance to male donors exhibited significantly greater female bias than more distant plants. The amount of female bias was positively correlated with the seed set of inflorescences. Hand pollination of stigmas resulted in approximately 100-fold higher stigmatic pollen loads than wind-pollinated stigmas and produced exceptionally female-biased progenies (female frequency = 0.96). Our results are the first to demonstrate a functional relation between stigmatic pollen capture, seed set, and sex ratio and suggest that certation can contribute towards female-biased sex ratios in dioecious plants.     

Sex determination and differentiation in Asparagus officinalis L.

The paper summarizes the coordinated researches conducted by three Italian groups in the area of sex determination and differentiation in the dioecious species Asparagus officinalis. Morphological evidence indicates that sex differentiation in Asparagus consists essentially of selective abortion of gynoecium or androecium of initially hermaphroditic floral primordia occurring in genotypically determined male and female individuals. Abortion occurs in pollen-mother cells and anthers in females and in megaspore-mother cells but not in the vegetative tissues of the ovary in males. The differential developmental pathway is accompanied by changes in relative abundance of auxin and cytokinins. The genetic ssytem controlling abortion of male or female organs is apparently monogenic (possibly a bipartite gene) with factor(s) associated with the homomorphic chromosome pair L5. Other genes influence the development of reproductive structures as indicated by the presence of genetic factors controlling stylar growth in male plants. The presence of extensive polymorphism in isoenzyme and DNA restriction fragment length patterns (RFLP) allows the search for markers associated with ‘sex genes’: a locus encoding a malic dehydrogenase (MDH) isoenzyme has been found about 20 cM from sex genes implying that chromosomes in which sex factors are located could pair and recombine. Searches for messages specifically expressed in reproductive structures were conducted by 2D-electrophoresis of existing and newly synthesized polypeptides or of in vitro translation products of poly(A) ⁺RNA from male and female flowers and by isolating specific monoclonal antibodies against sex specific floral antigens.     

Life‐history trade‐offs promote the evolution of dioecy

Most dioecious plants are perennial and subject to trade‐offs between sexual reproduction and vegetative performance. However, these broader life‐history trade‐offs have not usually been incorporated into theoretical analyses of the evolution of separate sexes. One such analysis has indicated that hermaphroditism is favoured over unisexuality when female and male sex functions involve the allocation of nonoverlapping types of resources to each sex function (e.g. allocations of carbon to female function vs. allocations of nitrogen to male function). However, some dioecious plants appear to conform to this pattern of resource allocation, with different resource types allocated to female vs. male sex functions. Using an evolutionarily stable strategy approach, we show that life‐history trade‐offs between sexual reproduction and vegetative performance enable the evolution of unisexual phenotypes even when there are no direct resource‐based trade‐offs between female and male sex functions. This result might help explain the preponderance of perennial life histories among dioecious plants and why many dioecious plants with annual life histories have indeterminate growth with ongoing trade‐offs between sexual reproduction and vegetative growth.     

Sex-specific cell division during development of unisexual flowers in the dioecious plant Silene latifolia

We analyzed cell division patterns during the differentiation of unisexual flowers of the dioecious plant Silene latifolia using in situ hybridization with histone H4 and cyclin A1 genes. The gene expression patterns indicated that the activation of cell divisions in whorls 3 and 4 was reversed in young male and female flower buds. During maturation of flower buds, a remarkable reduction in cell division activity occurred in the male gynoecium primordium and female stamen primordia. Our analyses showed that differential activation and reduction of cell division strongly correlated with sex-specific promotion and cessation in the sex differentiation of unisexual flowers.     

Master Regulatory Genes, Auxin Levels, and Sexual Organogeneses in the Dioecious Plant Mercurialis annua

In Mercurialis annua L. (2n = 16) genes for sex determination are considered as major regulator genes controlling stamen and ovary development and sexual phenotypes. After stamen induction, sterility determinants control sporogenous tissue and pollen formation. Moreover, exogenous auxins are able to induce male flowers on female plants. In order to verify if sex and sterility genes have an effect on indole-3-acetic acid (IAA) contents of these plants, various wild or genetically constructed strains were assayed. The IAA levels of their apices were determined by HPLC followed by gas chromatography, selected ion monitoring, mass spectrometry. Results show that high auxin levels are linked to male phenotypes. The genes inducing maleness and the determinants of restored male fertility appear to control and modulate the IAA content. Close correspondence between the number of these dominant genes and IAA levels was established. A final hypothesis about the control of sexual specialization by phytohormones induced by the presence of these genes is discussed.     

Construction of male and female PAC genomic libraries suitable for identification of Y-chromosome-specific clones from the liverwort, Marchantia polymorpha

Unlike higher plants, the dioecious liverwort, Marchantia polymorpha, has uniquely small sex chromosomes, with X chromosomes present only in female gametophytes and Y chromosomes only in male gametophytes. We have constructed respective genomic libraries for male and female plantlets using a P1-derived artificial chromosome (pCYPAC2). With an average insert size of approximately 90 kb, each PAC library is estimated to cover the entire genome with a probability of more than 99.9%. Male-specific PAC clones were screened for by differential hybridization using male and female genomic DNAs as separate probes. Seventy male-specific PAC clones were identified. The male specificity of one of the clones, pMM4G7, was verified by Southern hybridization and PCR analysis. This clone was indeed located on the Y chromosome as verified by fluorescence in situ hybridization (FISH). This result shows that the Y chromosome contains unique sequences that are not present either on the X chromosome or any of the autosomes. Thus, the respective male and female libraries for M. polymorpha offer an opportunity to identify key genes involved in the process of sex differentiation and this unique system of sex determination.     

Sex ratio and gender stability in the dioecious plants of Israel

The hypothesis that in dioecious plants genders are usually stable and that the sex ratio is 1:l was tested in the dioecious species of Israel. The flora of Israel comprises some 2500 wild species, among which 43 are dioecious, one gynodioecious and one androdioecious. Forty-one out of these 45 species were examined during 1995-97. These species were examined in 120 populations comprising 7019 individuals, at 81 different localities. The majority of the species (33/41) manifested the expected 1:1 sex ratio. Of the eight species deviating from this ratio, a significant bias was repeatedly scored in five of them. Three other species of the willow family manifested sometimes strongly biased sex ratios or even monomorphic (unisexual) stands, apparently as a result of extensive vegetative cloning. The stability of the male and female morphs was also examined in the 41 species. Only eight individuals (from seven species) out of the 7019 individuals surveyed (0.1%) were found to be bisexual. In addition, in 27 populations belonging to 17 species, 909 males were tagged and revisited in fruiting time, and only three of them (0.3%) were found to set fruits. In addition not a single case of gender reversal between seasons was found. The field survey suggests that in dioecious plants sex determination and expression are usually effectively maintained. The very slight deviations from the 1:1 sex ratio and from the distinct male and female morphs suggest that sexual dimorphism in dioecious plants is usually stabile.     

Gender in plants: sex chromosomes are emerging from the fog

Although most plants have flowers with both male and female sex organs, there are several thousands of plant species where male or female flowers form on different individuals. Surprisingly, the presence of well-established sex chromosomes in these dioecious plants is rare. The best-described example is white campion, for which large sex chromosomes have been identified and mapped partially. A recent study presented a comprehensive genetic and physical mapping of the genome of dioecious papaya. It revealed a short male specific region on the Y chromosome (MSY) that does not recombine with the X chromosome, providing strong evidence that the sex chromosomes originated from a regular pair of autosomes. The primitive papaya Y chromosome thus represents an early event in sex chromosome evolution. In this article, we review the current status of plant sex-chromosome research and discuss the advantages of different dioecious models.