Esterase activity and isozymes during the ontogeny of stamens of male fertile lycopersicon esculentum mill., A male sterile stamenless-2 mutant and the low temperature-reverted mutant

A comparative study of the activity and isozymes of esterase during the ontogeny of stamens of normal male fertile (MF) Lycopersicon esculentum, a male sterile stamenless-2 (sl-2/sl-2) mutant (MS), and the low temperature reverted mutant was conducted. In MF and MS stamens, a major isozyme associated with tapetum development was present at early stages whereas another isozyme related to pollen formation was observed at later stages of development. There was a difference, however, in the timing of the appearance of these isozymes in MF and MS stamens. Also, the number and intensity of most bands, and the overall and specific activity of esterase was higher in MF than in MS stamens. The isozyme pattern, the number and intensity of bands and the overall activity of esterase was comparable in MF and MS-reverted stamens from low temperature grown plants. Male fertile stamens from low temperature-grown plants contained lower esterase activity than those grown in the greenhouse. The results are discussed with respect to the mechanism of male sterility in the sl-2/sl-2 mutant of tomato.     

Transcriptomic analysis of differentially expressed genes in flower-buds of genetic male sterile and wild type cucumber by RNA sequencing

Cucumber (Cucumis sativus L.) pollen development involves a diverse range of gene interactions between sporophytic and gametophytic tissues. Previous studies in our laboratory showed that male sterility was controlled by a single recessive nuclear gene, and occurred in pollen mother cell meiophase. To fully explore the global gene expression and identify genes related to male sterility, a RNA-seq analysis was adopted in this study. Young male flower-buds (1–2 mm in length) from genetic male sterility (GMS) mutant and homozygous fertile cucumber (WT) were collected for two sequencing libraries. Total 545 differentially expressed genes (DEGs), including 142 up-regulated DEGs and 403 down-regulated DEGs, were detected in two libraries (Fold Change ≥ 2, FDR < 0.01). These genes were involved in a variety of metabolic pathways, like ethylene-activated signaling pathway, sporopollenin biosynthetic pathway, cell cycle and DNA damage repair pathway. qRT-PCR analysis was performed and showed that the correlation between RNA-Seq and qRT-PCR was 0.876. These findings contribute to a better understanding of the mechanism that leads to GMS in cucumber.     

The causes of genetic male sterility in 3 soybaen lines

Summary The cause of male sterility in 3 soybean lines, TGM 103-1, N-69-2774 and TGM 242-4 was studied. In TGM 103-1, which was both male and female sterile, two different abnormalities were associated with sterility. Precocious movement of a few chromosomes at the metaphase I stage resulted into the production of non-functional pollen while cells which underwent apparent normal meiotic division had disintergration of the tapetal cell wall immediately after the free microspore stage leading to the starvation and subsequent death of the developing microspores. In lines N-69-2774 and TGM 242-4, both of which were partially sterile, male sterility resulted from a failure of cytokinesis after the telophase II stage. Meiosis proceeded normally but the 4 microspores after telophase II failed to separate into pollen grains and degenerated thereafter.     

Genetics and development of morphological and physiological characters of male sterility in Oenothera

Summary Male sterility in Oenothera is influenced by two nuclear genes,fr andster. Their function is independent of the plastomes. Development of anthers, fertile and sterile male, was studied by electron microscopy and histochemical methods. Both genes act on lipid metabolism but at different developmental stages. Infr/fr homozygotes the disturbance is expressed as a lack of sporopollenin in the exine, while amorphous lipid material is deposited in the loculus. Inster/ster homozygotes sporopollenin is formed normally in the endexine but the paracristalline structure of the ektexine is missing. In both mutants the disturbance leads to complete destruction of the pollen grain. The deviation from fertile pollen development is correlated with abnormalities of the tapetum and outer cell layers of the anther wall.     

拟南芥雄性不育突变体ms1502的遗传及定位分析

通过EMS诱变、背景纯化与遗传分析,从拟南芥（Arabidopsis thaliana）中筛选到了一棵隐性单基因控制的雄性不育突变体ms1502。细胞学观察发现,突变体在小孢子从四分体释放出后花药绒毡层过早衰亡,小孢子的内容物不正常地凝聚,最终无法形成正常的花粉粒。利用图位克隆的方法对该基因MSl502进行了定位,结果表明MS1502位于第4条染色体上分子标记F25124和T12H20之间105kb区间内。目前该区间内尚未见到花药发育必需基因（不育基因）的报道,因此MS1502是一个控制花粉发育的新基因。     

In-situ seed production after pollination with in-vitro-matured,isolated pollen

Immature tobacco (Nicotiana tabacum L.) pollen has been isolated from anthers in three distinct stages of development, including the microspore stage. In in-vitro cultures, fully functional, mature pollen was obtained. In a germination medium, this pollen produced pollen tubes. After application to stigmas in situ, the in-vitro-matured pollen fertilized ovules, and seeds were produced. Genetic tests with seedlings obtained from pollinations with in-vitro-matured pollen from a transgenic plant revealed normal Mendelian segregation of two marker genes, the neomycin-phosphotransferase II gene and the nopaline-synthase gene. These results are of interest with respect to the control of self-incompatibility, cytoplasmic male sterility and pollen-allergen formation, and it offers an alternative route for gene transfer in those plants which cannot be regenerated in vitro.Abbreviations cms cytoplasmic male sterility; AMGLU, MS, M2S, MR26 - GK culture media, see Material and methods     

Flower production,male sterility and berry setting in andigena potato

Summary Unlike tuberosum, andigena potato germ plasm exhibits a high degree of genetic variation in morphological, biochemical and reproductive traits. Sixty-five percent of the 565 genotypes comprising 145 accessions of Solanum tuberosum ssp. andigena obtained from Argentina, Bolivia, Chile, Colombia and Peru remain totally vegetative and never develop any floral bud when cultivated in northern India. In 18% of genotypes, the floral buds develop but they drop off from the plants prematurely. Thus, 83% of genotypes do not develop mature flowers. The frequency of such genotypes is maximum in the Bolivian genotypes. Whereas 17% of genotypes produce mature flowers, only 2% develop berries. The highest proportion of floral bud formation and their subsequent development and differentiation into mature flowers occur in Peruvian and Colombian genotypes. Partial to high male sterility occurs in 93% of the flowering genotypes; their pollen sterility ranges from 15% to 91%. Seven percent of the flowering genotypes are completely pollen sterile. The male sterility is expressed variously, ranging from structural to sporogenous types. The floral bud formation, its development and retention to maturity, pollen and ovule functionability and fruit development are under the control of a large number of genes, most of which are unlinked and independent. Many of these genes are polygenic in nature.     

Patterns of male sterility within and among populations of the distylous shrub Erythroxylum havanense (erythroxylaceae)

Distyly, a reproductive system characterized by the presence of long-styled (thrum). and short-styled (pin) individuals within a population, has been repeatedly used as a model for the study of the evolution of the reproductive systems in plants. Erythroxylum havanense is a distylous species in which most thrum plants fail to develop a fertile androecium, thus behaving as male-sterile or partially male-sterile plants. Short-styled (thrum) individuals have an increased performance as female parents, thereby compensating for their loss of male fitness. Previous studies of populations within close proximity to each other suggest that E. havanense may be involved in a process of gender specialization in which, unlike other heterostylous species, thrum plants are specializing as females and pins (long-styled) as males. In this paper we describe more general patterns of male sterility, one of the first steps in the evolution of gender specialization, among populations of the distylous shrub Erythroxylum havanense. Pollen germination differed among populations (range 0.52 ± 0.03 to 0.06 ± 0.04), and between morphs. Pollen from pin plants was almost two times (1.89) as fertile as that from thrums (0.36 ± 0.03 and 0.19 ± 0.03, pin and thrums respectively). Thrums were significantly more male sterile in four out of five populations. The population where differences between the floral morphs were not apparent showed the lowest levels of pollen fertility. Accordingly, our results indicate that populations of E. havanense show marked differences in pollen fertility and higher male sterility associated with the thrum morph. We hypothesize that differences between morphs could be explained if restorers of male sterility are linked to the distyly haplotype, while differences in genes associated with male sterility could explain the variation among populations. Overall, the prevalence of thrum-biased male sterility across populations suggests that E. havanense is subject to a process of gender specialization.     

Trihalogenated methylsulfonamides as specific male gametocides

A new class of male gametocide is described: trihalomethylsulfonamides, the most active example of this class being trifluoromethylsulfonamide (TFMSA). TFMSA induces male sterility, specifically, without detectable effects on other plant functions. Male sterility induction in maize (Zea mays, gaspe flint) required minimally 200 g TFMSA per plant and this rate was used in a metabolite sampling time-course experiment to determine the earliest detectable change in metabolites of developing florets. Metabolites profiled were amino acids, callose, fatty acids, flavones, phenylpropenoids, sporopollenin and starch, all of which are related to successful pollen development. Changes in proline and starch were the earliest statistically significant differences observed between florets of control plants and TFMSA-treated plants. These metabolic differences were observed before symptoms of pollen failure were evident. In subsequent experiments, transient increases in glume proline and decreases in anther proline were linearly related to sub-effective rates of TFMSA (0, 25, 50, 75 and 100 g plant^–1). Increases in glume proline faded during development whereas decreases in anther proline linearly related to TFMSA rate became more prominent. Changes in all other metabolites profiled were not linearly related to TFMSA rate. Related experiments showed that florets from TFMSA-treated plants were not capable of converting ¹⁴C-glutamate to ¹⁴C-proline, and the anther transport capacity of ¹⁴C-proline in TFMSA-treated plants was significantly reduced. It is inferred that TFMSA induces male sterility by interfering with the transport of proline from the site of synthesis to the site of accumulation, resulting in feedback inhibition of proline biosynthesis, ultimately starving the developing anther of proline.     

Tapetum-specific expression of harpinPss causes male sterility in transgenic tobacco

Harpin, an elicitor molecule of bacterial origin induces hypersensitive response (HR) in non-host plants. In an attempt to induce male sterility, harpin was tagged with a signal peptide and expressed downstream to tapetum-specific TA29 promoter resulting in extracellular secretion, subsequent degeneration of tapetum and development of male sterility in tobacco. Putative transgenics were analyzed by PCR amplification of transgene, semiquantitative RT-PCR analysis from total RNA extracts from anther tissue with transgene specific probe, Western blotting using polyclonal antibody raised against harpin, by transmission and scanning electron microscopy, and by confocal microscopy of anthers and pollen at various stages of development. Varying degrees of male sterility (30–100 %) was observed with plants showing complete and partial male sterility as well as several morphological variations were seen especially in leaves and flowers. Further, some of the transgenics showed un-induced of HR-like local lesions in the vegetative tissues. Harpin_Pss got deposited on the pollen grains upon tapetal degeneration resulting in significant alterations in the morphology of pollen cell wall. However, megagametogenesis was not affected in complete and partial male sterile plants and female gametes were completely fertile. The complete male sterility was attributed to premature tapetal cell death due to sufficient extracellular harpin_Pss accumulation whereas insufficient protein content might be the reason for partial male sterility. These findings indicate the possible use of cytotoxic harpin_Pss for the development of male sterile plants.