桃花粉育性与花药颜色的关系及其SSR分子标记

以李属桃亚属的637份桃品种种质和栽培种瑞光19×Summergrand杂交的138株F1群体为试材,对桃花粉育性与花药颜色的关系及其SSR分子标记进行研究。结果表明:在所有品种中,橘红花药最多,占总份数的91.2%,其中绝大部分花粉可育;橘黄和黄色花药的种质数量次之;白色和浅褐花药的种质最少,且均表现为花粉不稔;整体表现为红色和黄色花药与花粉可育具有较强的正相关性。本试验从122对SSR引物中筛选出2对与花粉育性性状连锁的标记CPDCT013和CP-SCT012,根据这2个标记参考整合参考图谱的位置,将控制花粉育性的基因定位在桃第6条染色体上端。对已经报道的2个桃花粉育性标记CPPCT004和NNJ-I以及第6条连锁群的其他SSR位点在花粉育性不同的24个品种上验证,结果表明只有UDP96-001(125bp)可以用于桃花粉育性的分子标记辅助育种。     

Regeneration of doubled haploid plants by androgenesis of cucumber (<Emphasis Type="Italic">Cucumis sativus </Emphasis>L.)

Six experiments (including pretreatment, embryonic callus induction media, preculture conditions, embryo induction media, embryo germination media, and genotypic effects) were conducted to develop an efficient cucumber (Cucumis sativus L., 2n = 2x = 14) anther culture protocol. Pretreatment and embryo induction were key factors for successful anther culture. Suitable temperature stress depended on the ecotype, i.e., cucumbers from cold areas responded well to cold shock whereas those from temperate areas responded well to heat treatment. The best medium for embryonic callus induction was MS medium supplemented with 4.44 μM BA, 2.26 μM 2, 4-D, 4.64 μM KIN, 3% sucrose and 0.8% agar. For embryo induction, MS medium supplemented with 0.54 μM NAA, 13.32 μM BA, 3% sucrose and 0.8% agar was optimal, and for embryo germination MS medium containing 2.22 μM BA, 6% sucrose and 1.2% agar was best. Using this protocol, we produced callus from 16 genotypes and regenerated plants from three of 20 evaluated. Three embryos per anther and 42 DH per 45 anthers (93% success) were obtained for cv. Ningjia No. 1, which was an improved result over a previous report. The origin of regenerants from microspores was determined by cytological, morphological and AFLP analyses.     

Sucrose and starch catabolism in the anther of <Emphasis Type="Italic">Lilium</Emphasis> during its development: a comparative study among the anther wall,locular fluid and microspore/pollen fractions

In order to better understand the various pathways of sucrose and starch catabolism in the anther of lily (Lilium hybrida var. “Enchantment”), invertase (EC 3.2.1.26) and amylase (EC 3.2.1.1, EC 3.2.1.2) activities were measured separately in different fractions (anther wall, locular fluid and microspore/pollen) and correlated with the sugar content during anther development. Our findings showed significant differences among the fractions analyzed, suggesting that the regulation of sucrose and starch catabolism could follow distinct pathways in each fraction. Glucose and fructose amounts progressively decreased from anther wall to fluid and from fluid to microspore/pollen. Thus, the developing pollen could act as a sink for the carbohydrates that reach the anther. In this sense, cell wall-bound invertases seem to play a major role in soluble sugar partitioning in the different fractions of the anther. Sucrose concentration was found to be substantially higher in the locular fluid than in the other fractions, indicating a probable site for storage. On the other hand, the anther wall tissues could have a buffering function, storing nutrient surplus in starch grains and thus regulating the availability of soluble sugars in the whole anther. All these results proved the advantages of the experimental model proposed here, as well as its usefulness to investigate sugar metabolism in Lilium anthers.     

RNAi-mediated male sterility of tobacco by silencing TA29

The superior performance of F₁ hybrids has a significant impact on agricultural productivity. For commercial application, the availability of an efficient system for obtaining male-sterile lines of crops is an essential prerequisite. Here we have investigated the use of RNA interference (RNAi) technology to silence a male-specific gene in the model host tobacco. TA29 is expressed exclusively in anthers at the time of microspore development. About 10 out of 13 tobacco lines transformed with a hairpin RNAi construct containing TA29 sequences were male sterile. Transgenic plants were phenotypically indistinguishable from non-transgenic plants. At the anthesis stage, pollen grains from transgenic, male-sterile plants were aborted and lysed in comparison to the round and fully developed pollen in non-transgenic plants. Microscopic analysis of anthers showed selective degradation of tapetum in transgenic plants with no microspore development. One week after self-pollination, the ovules of non-transgenic plants were double the size of those in transgenic plants, due to successful self-fertilization. Male sterile transgenic plants set seed normally, when cross-pollinated with pollen from non-transgenic plants, confirming no adverse effect on the female parts of the flower. These results show that silencing of male-specific genes by RNAi is potentially a useful tool for generating male-sterile lines for producing hybrid seed.     

Rate of resistance evolution and polymorphism in long‐ and short‐lived hosts

Recent theoretical work has shown that long‐lived hosts are expected to evolve higher equilibrium levels of disease resistance than shorter‐lived hosts, but questions of how longevity affects the rate of resistance evolution and the maintenance of polymorphism remain unanswered. Conventional wisdom suggests that adaptive evolution should occur more slowly in long‐lived organisms than in short‐lived organisms. However, the opposite may be true for the evolution of disease‐resistance traits where exposure to disease, and therefore the strength of selection for resistance increases with longevity. In a single locus model of innate resistance to a frequency‐dependent, sterilizing disease, longer lived hosts evolved resistance more rapidly than short‐lived hosts. Moreover, resistance in long‐lived hosts could only be polymorphic for more costly and more extreme resistance levels than short‐lived hosts. The increased rate of evolution occurred in spite of longer generation times because longer‐lived hosts had both a longer period of exposure to disease as well as higher disease prevalence. Qualitatively similar results were found when the model was extended to mortality‐inducing diseases, or to density‐dependent transmission modes. Our study shows that the evolutionary dynamics of host resistance is determined by more than just levels of resistance and cost, but is highly sensitive to the life‐history traits of the host.     

Cotton LIM domain‐containing protein GhPLIM1 is specifically expressed in anthers and participates in modulating F‐actin

As one form of actin binding protein (ABP), LIM domain protein can trigger the formation of actin bundles during plant growth and development. In this study, a cDNA (designated GhPLIM1) encoding a LIM domain protein with 216 amino acid residues was identified from a cotton flower cDNA library. Quantitative RT‐PCR indicated that GhPLIM1 is specifically expressed in cotton anthers, and its expression levels are regulated during anther development of cotton. GhPLIM1:eGFP transformed cotton cells display a distributed network of eGFP fluorescence, suggesting that GhPLIM1 protein is mainly localised to the cell cytoskeleton. In vitro high‐speed co‐sedimentation and low co‐sedimentation assays indicate that GhPLIM1 protein not only directly binds actin filaments but also bundles F‐actin. Further biochemical experiments verified that GhPLIM1 protein can protect F‐actin against depolymerisation by Lat B. Thus, our data demonstrate that GhPLIM1 functions as an actin binding protein (ABP) in modulating actin filaments in vitro, suggesting that GhPLIM1 may be involved in regulating the actin cytoskeleton required for pollen development in cotton.     

不结球白菜雄性不育系及其保持系花药发育的细胞学观察

以不结球白菜（Brassica campestris ssp．chinensis Makino）雄性不育系及其保持系为试验材料,选择不同发育阶段的花蕾,取其花药,制成石蜡切片和超薄切片,经染色后在电子显微镜下观察。结果表明,不结球白菜雄性不育系与保持系的花药发育有明显的不同：不育系花药发育受阻于花粉母细胞分化期,形成1～3个药室,并形成正常的四分体小孢子,此时细胞组织逐步解体,形成空腔花药;最后向内皱缩;保持系花粉母细胞能形成正常的四分体,进而形成小孢子,最终形成充满正常花粉粒的花药。     

Cytological analysis and genetic control of rice anther development

Microsporogenesis and male gametogenesis are essential for the alternating life cycle of flowering plants between diploid sporophyte and haploid gametophyte generations.Rice (Oryza sativa) is the world's major staple food,and manipulation of pollen fertility is particularly important for the demands to increase rice grain yield.Towards a better understanding of the mechanisms controlling rice male reproductive development,we describe here the cytological changes of anther development through 14 stages,including cell division,differentiation and degeneration of somatic tissues consisting of four concentric cell layers surrounding and supporting reproductive cells as they form mature pollen grains through meiosis and mitosis.Furthermore,we compare the morphological difference of anthers and pollen grains in both monocot rice and eudicot Arabidopsis thaliana.Additionally,we describe the key genes identified to date critical for rice anther development and pollen formation.     

Correlation of sequential floral and male gametophyte development and preliminary results on anther culture in <Emphasis Type="Italic">Opuntia ficus-indica</Emphasis>

Before approaching anther culture as a tool to trigger an androgenic response in a new species, it is advisable to characterize and correlate flower and male gametophyte development to enable reproducible identification of the appropriate starting material. Buds and flowers of Opuntia ficus-indica cv. Gialla were classified in eight stages according to their total length at the earlier stages and the length of the corolla in flowers with emerging sepals. Due to the low condensation of chromatin in the microspore nucleus as well as in the vegetative nucleus of the bi- and tricellular pollen along with the high autofluorescence of the intricate exine, DAPI staining turned out not to be feasible in this species. Therefore an approach based on light-microscopy observation of semithin sections was used. These sections were stained with toluidine blue for general structure recognition and I₂KI to study starch deposition. Correlations were made between the sequential floral and male gametophyte development. Using this approach we determined the timing of pollen formation and observed that pollen development is impaired in plants producing seedless fruits. Furthermore, anther culture was carried out with anthers collected from flower buds at stages 2 and 3. Most of the anthers produced callus, however no regeneration was obtained.