棉花花粉发育过程中同工酶和蛋白质的研究

利用超薄胶电泳技术对陆地棉花粉发育各期的酯酶和过氧化物酶的同工酶以及蛋白质的种类及活性变化进行研究。结果发现：在花粉第一次有丝分裂结束之前,存在大量的低分子量酯酶和过氧化物酸酶,花粉成熟后又新出现一些大分子量酯酶和过氧化物酶在花粉第一次有线四分体阶段,了一种特异蛋白,推测具有调控作用。在有丝分裂结束后,出现了一些新的蛋白质,这些蛋白质存在的时间很短。之后大量蛋白质开始合成,并在数量和种类上有逐渐增     

华北落叶松花粉发育过程中的钙动态分布

运用焦锑酸钾沉淀法研究了华北落叶松(Larixprincipis-rupprechtiiMayr)小孢子发育过程中不同阶段Ca2 的分布情况。减数分裂时期,小孢子囊壁表皮和中层细胞的细胞壁及细胞间隙Ca2 分布较多,绒毡层只有外切向面的细胞膜有Ca2 分布,小孢子母细胞的各部位则很少有Ca2 ;四分体时期,包围四分小孢子的胼胝质壁上有大量的Ca2 分布,在四分孢子壁上也有较多沉淀;游离小孢子时期,钙离子在小孢子壁的分布较四分体时期有所减少,而到花粉成熟时又逐渐增多;从四分体到花粉成熟,乌氏体周围的Ca2 有增多的趋势。对四分体外壁Ca2 的大量分布与花粉壁的形成及信号物质在花粉表面贮存的关系,以及小孢子囊的外壁、绒毡层和乌氏体在Ca2 向花粉运输中所起的作用进行了讨论。     

华北落叶松花粉发育过程中的钙动态分布

运用焦锑酸钾沉淀法研究了华北落叶松(Larix principis-rupprechtii Mayr)小孢子发育过程中不同阶段Ca2 的分布情况.减数分裂时期,小孢子囊壁表皮和中层细胞的细胞壁及细胞间隙Ca2 分布较多,绒毡层只有外切向面的细胞膜有Ca2 分布,小孢子母细胞的各部位则很少有Ca2 ;四分体时期,包围四分小孢子的胼胝质壁上有大量的Ca2 分布,在四分孢子壁上也有较多沉淀;游离小孢子时期,钙离子在小孢子壁的分布较四分体时期有所减少,而到花粉成熟时又逐渐增多;从四分体到花粉成熟,乌氏体周围的Ca2 有增多的趋势.对四分体外壁Ca2 的大量分布与花粉壁的形成及信号物质在花粉表面贮存的关系,以及小孢子囊的外壁、绒毡层和乌氏体在Ca2 向花粉运输中所起的作用进行了讨论.     

桔梗花粉母细胞减数分裂及雄性败育的细胞生理学研究

以桔梗不育系PA及其保持系PB为试验材料,采用石蜡切片和改良苯酚品红染色压片法对花粉母细胞减数分裂和雄配子体发育过程进行比较,探讨桔梗雄性不育系小孢子形成过程和败育发生的细胞生理学机理。结果表明:(1)桔梗保持系PB花粉母细胞减数分裂的细胞质分裂为同时型,同一花药减数分裂较同步;在中期Ⅰ和中期Ⅱ,少数细胞中可见赤道板外染色体;四分体以四面体为主,成熟花粉粒为二核花粉。(2)不育系PA花粉母细胞减数分裂后期Ⅰ开始出现异常,表现为细胞质形态改变,末期Ⅱ之后细胞质不能分裂,形成异常四分体,胼胝质壁不能溶解,四分体难以释放出游离小孢子而被降解,导致败育。(3)在发育过程中,桔梗不育系花蕾游离脯氨酸、可溶性蛋白含量低于保持系,而SOD活性、丙二醛含量均高于保持系。(4)桔梗不育系PA及其保持系PB花粉母细胞减数分裂和雄配子体发育过程存在明显差异,桔梗雄性败育过程大体可分为4个阶段,即后期Ⅰ细胞质异常、末期Ⅱ之后细胞质不能分裂、四分体难以释放游离小孢子、四分体被降解仅残留碎片。研究认为,桔梗不育花蕾(开花前)生长发育过程中,体内活性氧代谢紊乱、丙二醛积累及游离脯氨酸等"物质代谢损亏"可能是引起桔梗雄性败育的原因。     

萝卜贮藏期可溶性蛋白及同工酶研究

试验以４个萝卜（Ｒａｐｈａｎｕｓ ｓａｔｉｖｕｓ Ｌ．）品种为试标,研究了贮藏期可溶性蛋白含量及ＰＡＧＥ蛋白图谱、酯酶和淀粉酶的同工酶谱。试验结果表明,在贮藏期蛋白浓度随贮存时间延长而下降,ＰＡＧＥ蛋白谱带显示不仅有品种之间蛋白种类的差异而且有贮藏期不同阶段的差异,淀粉酶同工酶谱显示随贮藏时间延长,酶活性下降,同工酶谱带增加,酯酶在贮藏过程中酶活性呈现先升高再下降然后再升高的变化,同工酶谱带贮藏中     

辣椒花药培养胚状体发生的组织学和细胞学研究

采用荧光显微镜、扫描电镜和透射电镜技术．系统研究了辣椒花药培养胚状体发生的组织学和细胞学变化特征。辣椒单个花药中花粉发育具有强烈的不同步性。随着培养时期的变化．不同时期花粉的百分率也发生变化。处于单核靠边期的小孢子培养以后按两种发育途径之一进行发育。在多数情况下,孢子体不对称分裂,产生典型双核花粉。胚性花粉粒是由营养核的重复分裂形成的。当小孢子从四分体中释放出来．特殊类型的外壁已经形成。在随后的花粉发育过程中．小孢子体积增大,外壁继续加厚。培养24h后,小孢子体积增大。胚性发生的小孢子表现出两种不同的形态变化。当胚状体发育到心形胚时．胚状体的表皮细胞排列规则。用光学和电子显微镜分析了小孢子胚状体形态形成过程．及胚状体诱导后细胞组织发生的一系列结构变化的时序性特征,这些变化主要影响质体、液泡室、细胞壁和细胞核,进一步分化的程序模拟合子胚的发育。     

羽叶薰衣草雄性不育的细胞学研究

用光镜和电镜观察羽叶薰衣草(Lavandula pinnata L.)雄性不育小孢子发育过程的细胞形态学特征.结果表明:羽叶薰衣草花药4枚,每枚花药通常具4个小孢子囊.花药壁发育为双子叶型,从外向内分为表皮、药室内壁、中层和绒毡层4层细胞.减数分裂形成的四分体为四面体及十字交叉型.小孢子的发育过程可分为造孢细胞期、减数分裂时期、小孢子发育早期、小孢子发育晚期.未观察到二胞花粉期和成熟花粉期.羽叶薰衣草花粉败育主要发生在单核花粉时期,细胞内物质解体并逐渐消失变成空壳花粉或花粉皱缩变形成为各种畸形的败育花粉.在此之前小孢子的发育正常.羽叶薰衣草小孢子不育机制体现在绒毡层过早解体、四分体时期以后各细胞中线粒体结构不正常、胼胝质壁与小孢子母细胞脱离、花药壁细胞中淀粉出现时间异常等. 壁发育为双子叶型,从外向内分为表皮、药室内壁、中层和绒毡层4层细胞.减数分裂形成的四分体为四面体及十字交叉型.小孢子的发育过程可分为造孢细胞期、减数分裂时期、小孢子发育早期、小孢子发育晚期.未观察到二胞花粉期和成熟花粉期.羽叶薰衣草花粉败育主要发生在单核花粉时期,细胞内物质解体并逐渐消失变成空壳花粉或花粉皱缩变形成为各种畸形的败育花粉.在此 前小孢子的发育正常.羽叶薰衣草小孢子不育机制体现在绒毡层过早解体、四分体时期以后各细胞中线粒体结构不正常、胼胝质壁与小孢子母细胞脱离、花药壁细胞中淀粉出现时间异常等. 壁发育为双子叶型,从外向内分为表皮、药室内壁、中层和绒毡层4层细胞.减数分裂形成的四分体为四     

辣椒雄性不育材料H9A小孢子败育机理

运用光学、电子显微技术及生化指标测定法, 对辣椒（Capsicum annuum）雄性不育材料H9A及其保持系H9B小孢子不同发育时期进行细胞形态学观察及生化特性分析。结果表明, 雄性不育材料H9A的小孢子败育发生在单核前期, 由于绒毡层细胞极度膨胀, 四分体受挤压后破裂并降解, 无法形成正常的单核花粉粒, 属于孢子体败育。不育材料花蕾中的过氧化物酶活性从四分体时期开始明显高于保持系, 保持系游离脯氨酸含量从单核小孢子期开始明显高于不育材料; 不育材料小孢子发育各时期的可溶性蛋白含量都明显低于保持系, 但丙二醛含量均高于保持系。     

辣椒雄性不育材料H9A小孢子败育机理

运用光学、电子显微技术及生化指标测定法, 对辣椒(Capsicum annuum)雄性不育材料H9A及其保持系H9B小孢子不同发育时期进行细胞形态学观察及生化特性分析。结果表明, 雄性不育材料H9A的小孢子败育发生在单核前期, 由于绒毡层细胞极度膨胀, 四分体受挤压后破裂并降解, 无法形成正常的单核花粉粒, 属于孢子体败育。不育材料花蕾中的过氧化物酶活性从四分体时期开始明显高于保持系, 保持系游离脯氨酸含量从单核小孢子期开始明显高于不育材料; 不育材料小孢子发育各时期的可溶性蛋白含量都明显低于保持系, 但丙二醛含量均高于保持系。     

湿地松雄性不育和可育系小孢子叶球形态和细胞发育动态变化

为了解湿地松‘松泰’小孢子叶球在发育过程形态是否有差异变化,明确其败育过程、败育方式及影响因素,为湿地松雄性不育品种利用和后期开展相关研究提供科学依据。该研究以‘松泰’s10败育系和s9可育系为材料,观察小孢子叶球形态发育变化,并对其小孢子叶球进行石蜡切片,在光学显微镜下观察小孢子发育过程。结果表明:s10败育系和s9可育系在小孢子母细胞减速分裂前无明显差异,小孢子叶球生长趋势也一致;四分体时期,s10小孢子细胞发育异常,小孢子叶球形态发育也出现异常,二者异常发育具有同步性;可育系从四分体到单核小孢子发育阶段的时间为5 d左右,而败育系持续发育长达20 d左右,持续时间为可育系的4倍。在此期间出现小孢子绒毡层细胞发育异常、降解缓慢,小孢子囊壁组织排列紊乱、降解延迟等现象,s10形成异常二核花粉,且无花粉散出。因此,推论s10小孢子败育的原因主要是小孢子囊壁细胞发育异常,其小孢子叶球形态异常,相对应的绒毡层在四分体时期发育异常,不能适时地分泌胼胝质酶来降解围绕着四分体的胼胝质壁,也不能适时地合成输送花粉形成所需能量物质,同时囊壁细胞出现降解延迟和层积,这一系列的异常变化导致不能形成正常四分体,从而使花粉败育。     

利用单花粉蛋白电泳技术对玉米花粉发育过程特异蛋白表达的研究

八十年代,Ｍｕｌｃａｈｙ（１９８１）与Ｃ．Ｇａ６（１９８６）成功地对单个花粉粒同工酶和蛋白质进行了等电聚集电析。而国内在这方面的工作至今尚未见报道。试验利用单花粉蛋白电泳技术结合显微镜观察对不同发育时期的单个玉米花粉进行了蛋白电泳研究,结果表明玉米花粉不同发育时期所表达的蛋白质具有明显的差异：一方面随发育时期的推移某些蛋白质含量呈递减或递增趋势;另一方面不同发育时期都有特异性蛋白合成或分解。可见,     

Distribution of insoluble polysaccharides, neutral lipids, and proteins in the developing anthers of Campsis radicans (L.) Seem. (Bignoniaceae)

In this study, distribution of polysaccharides, lipids, and proteins in the developing anthers of Campsis radicans (L.) Seem. was examined from sporogenous cell stage to mature pollen, using cytochemical methods. To detect the distribution and dynamic changes of insoluble polysaccharides, lipid bodies, and proteins in the anthers through progressive developmental stages, semi-thin sections of anthers at different developmental stages were stained with periodic-acid-Schiff (PAS) reagent, Sudan black B, and Coomassie brilliant blue, respectively, and examined under light microscope. Ultrastructural observations with TEM were also carried out to determine the storage form of starch in the connective tissue, and storage form of lipids in the tapetal cells. In sporogenous cell stage, anther wall contains numerous insoluble polysaccharides. However, from the sporogenous cell stage to the vacuolated microspore stage, the amount of insoluble polysaccharides in the anther wall decreases gradually. At bicellular pollen stage, tapetum degenerates completely and polysaccharides are not seen in the anther wall. Lipid bodies are observed in the cytoplasm of both middle layer and tapetal cells at tetrad stage, whereas they disappear in the vacuolated microspore stage. Compared with polysaccharides, proteins are limited in the anther wall at early stages of development. During pollen development, polysaccharides, proteins, and lipid bodies are scarce in the cytoplasm of sporogenous cells, but their amount increases at premeiotic stage. From tetrad stage to bicellular pollen stage, microspore cytoplasm contains variable amount of insoluble polysaccharide grains, lipid and protein bodies. At bicellular pollen stage, plentiful amount of starch granules are stored in the cytoplasm of the pollen grains. Proteins and lipid bodies are also present in the cytoplasm.     

麻疯树小孢子发育的研究

用透射电镜观察了麻疯树(Jatropha curcas L.)小孢子发育的超微结构。小孢子母细胞时期内质网和质体较多;减数分裂和四分体时期,细胞处于明显的代谢活跃状态,细胞器丰富,主要有内质网、线粒体、质体、高尔基体和球状体;在小孢子发育早期和晚期,线粒体和内质网仍较丰富;小孢子经过高度的不对称分裂后,形成较大的营养细胞和较小的生殖细胞,营养细胞中细胞器数量明显减少,含大量的淀粉和脂类物质,生殖细胞中脂类物质丰富;表皮、药室内壁和中层细胞在小孢子母细胞和四分体时期淀粉粒丰富,小孢子时期明显减少,绒毡层从小孢子母细胞至小孢子发育晚期的细胞器都很丰富,主要为内质网、质体和线粒体,为二胞花粉发育奠定基础。     

Electrophoretic Survey of Proteins and Esterase Isozyme in Wheat×Maize Progenies

The analysis of soluble proteins and esterase isozyme in F2 progeny grains from wheat (Triticum aestivum L. ) × maize (Zea mays L. ) crosses indicated that the electrophoretic pattern of proteins and esterase isozymes was extremely different from that of their parents. Protein variation was mainly concentrated in the high-molecular-weight-Glu (HMW-Glu) zone. There were 5 kinds of protein eleetrophoretic patterns in the analyzed grains. VIZ: maternal, additional, complementary, hybrid and omission type which accounted for 22.6%, 14.3%, 15.5%, 30. 9% and 16.7% of the total tested grains respectively. In the analysis of esterase pattern, some variations in progenies were also found. The variations of electrophoretic pattern of proteins and esterase isozyme indicated that a genetic material change in wheat chromosomes could be induced in the distant hybridization.     

枸杞花药发育过程中脂滴和淀粉粒的分布特征

宁夏枸杞(Lycium barbarurn L．)花药发育过程中,淀粉粒和脂滴两种营养物质的积累和分布具有一定的特点：在造孢细胞时期,药隔薄壁细胞,表皮和药室内壁细胞中开始积累淀粉粒,而造孢细胞、绒毡层细胞和中层细胞中则没有淀粉粒。在四分体时期,绒毡层细胞开始积累脂滴并且数量逐渐增加。到小孢子晚期,绒毡层细胞降解,内含脂滴流入药室中。在小孢子发育过程中既没有淀粉粒也没有脂滴积累,直到二胞花粉的大液泡消失后花粉粒中才开始积累脂滴,然后又开始出现淀粉粒。枸杞成熟花粉中的营养储存物是脂滴和淀粉粒。     

Pollen development in a submerged plant,Ottelia alismoides (L.) Pers. (Hydrocharitaceae)

Exine structure and its developmental program in a submerged plant,Ottella alismoides (L.) Per. were investigated with scanning and transmission electron microscopies. Verrucate protrusions initiate on microspore plasma membrane at early tetrad stage. The verrucate protrusions develop into spines during free microspore stage. A foot layer is formed by accumulation of lamellated structure. The pollen grains ofOttelia alismoides are inaperturate, not omniaperturate, because of the well-developed foot layer. The inaperturate pollen grains ofOttelia are characterized by the spinous protrusions and the granular foot layer.     

Cytochemistry of pollen development in Campsis radicans (L.) Seem. (Bignoniaceae)

In this study, cytochemical staining methods were used to follow the cytochemical modifications of microspore cytoplasm and sporoderm in Campsis radicans (L.) Seem. from tetrad stage to mature pollen. Flower buds were collected at different stages of development, and the anthers were fixed and embedded in Araldite. To make cytochemical observations under light microscope, semithin sections were cut and stained with different dyes. Cytochemical methods provided the opportunity to localize the reserve material in the microspore and pollen cytoplasm, to distinguish the different layers of the sporoderm, and to determine its chemical structure at different developmental stages. Microspore cytoplasm contains variable amounts of proteins, lipids, and insoluble carbohydrates at different stages of microsporogenesis. Sporoderm formation starts at tetrad stage by the formation of primexine and is completed at vacuolated microspore stage by the addition of sporopollenin from tapetum. During the vacuolization and enlargement of the microspores, the structure and the chemical composition of the exine are modified. The endexine becomes chemically different from the ectexine. The ectexine is composed of sporopollenin and a small amount of protein, whereas the endexine is composed of sporopollenin, proteins, and traces of polysaccharides.     

Ultrastructure of microsporogenesis and microgametogenesis in Campsis radicans (L.) Seem. (Bignoniaceae)

In the present study, microsporogenesis, microgametogenesis and pollen wall ontogeny in Campsis radicans (L.) Seem. were studied from sporogenous cell stage to mature pollen using transmission electron microscopy. To observe the ultrastructural changes that occur in sporogenous cells, microspores and pollen through progressive developmental stages, anthers at different stages of development were fixed and embedded in Araldite. Microspore and pollen development in C. radicans follows the basic scheme in angiosperms. Microsporocytes secrete callose wall before meiotic division. Meiocytes undergo meiosis and simultaneous cytokinesis which result in the formation of tetrads mostly with a tetrahedral arrangement. After the development of free and vacuolated microspores, respectively, first mitotic division occurs and two-celled pollen grain is produced. Pollen grains are shed from the anther at two-celled stage. Pollen wall formation in C. radicans starts at tetrad stage by the formation of exine template called primexine. By the accumulation of electron dense material, produced by microspore, in the special places of the primexine, first of all protectum then columellae of exine elements are formed on the reticulate-patterned plasma membrane. After free microspore stage, exine development is completed by the addition of sporopollenin from tapetum. Formation of intine layer of pollen wall starts at the late vacuolated stage of pollen development and continue through the bicellular pollen stage.