棉花细胞质雄性不育花药败育过程中内源激素的变化

利用酶联免疫检测（ELISA）技术,研究了棉花细胞质雄性不育系、保持系、恢复系和杂种一代花药在造孢细胞增殖期、花粉母细胞减数分裂期、四分体至小孢子释放期、花粉发育期和花粉成熟期内源吲哚乙酸（IAA）、赤霉素（GA₃）、玉米素核苷（ZR）和脱落酸（ABA）含量的动态变化。结果表明,在保持系、恢复系和杂种一代可育花药间内源激素含量差异不明显,但在可育与不育花药间差异显著。在IAA、GA₃和ZR含量上,不育花药低于可育花药,但在ABA含量上,不育花药高于可育花药,这种差异在花粉母细胞减数分裂时期达最大值。根据内源激素的功能推测,不育花药IAA含量过低使花药淀粉积累受阻,GA₃含量不足影响花粉母细胞和绒毡层细胞的膨大,过低的ZR含量使花粉母细胞不能分裂形成四分体,过高的ABA含量促进花粉母细胞退化和死亡。     

玉米CMS-S小孢子败育过程中的细胞程序性死亡

以玉米（Zea mays L.）S型细胞质雄性不育系(CMS-S)的一对近等基因系S-Mo17^Rf3Rf3 和S-Mo17^rf3rf3为材料，采用TdT介导的dUTP DNA末端标记（TUNEL）、细胞色素C免疫原位杂交和DNA寡聚核小体片段电泳等方法，分别在细胞学水平和DNA水平上研究了玉米CMS-S小孢子败育的细胞程序性死亡（PCD）过程。结果表明，在花粉母细胞减数分裂后的四分体解离时期，不育花药的绒粘层细胞较可育花药提前裂解；在不育系S-Mo17^rf3rf3花药和花粉S-rf3中均明显出现PCD过程的DNA片段化以及线粒体细胞色素C外渗的现象，证明了玉米CMS-S的花粉败育与花药绒粘层细胞的提前凋亡和小孢子细胞的程序性死亡有关。     

Characterization of cytoplasmic male sterility in Petunia hybrida and Zea mays. Localization and activity of cytochrome c oxidase

Summary Anthers of male fertile, cytoplasmic male sterile (cms), and restored male fertile Petunia hybrida, are analyzed for cytochrome c oxidase (cox) activity in subsequent stages of microsporogenesis, and compared with anthers of male fertile, cms-S and cms-C Zea mays. The cox activity is determined in anther extracts and cytochemically. In petunia anthers, the first differences in cox activity occur from meiosis onward. However, at these stages, the initial symptoms of degeneration are already apparent. It is suggested that the decline in enzyme activity of the cms petunia anthers is the result rather than the cause of the non-formation of functional pollen.In maize anthers, the cox activity of sterile-type anthers is reduced in comparison with fertile-type anthers from premeiosis onward. There are also consistent cytochemical differences in the mitochondrial organization of cox activity between pollen of cms-S and male ferile maize anthers. In fertile-type mitochondria, the DAB reaction product indicating cox activity is localized in the cristae and within the space between the outer and inner limiting membranes of the organelles. In mitochondria of pollen of cms-S maize, cox activity is only observed between the outer and inner membranes of the mitochondria. The biochemical and cytochemical differences are observed at stages of development at which no structural signs of degeneration are apparent. The results suggest that cms in maize correlates with deviations in cytochrome c oxidase activity.Abbreviations cox cytochrome c oxidase - DAB 3,3-diaminobenzidine     

番茄雄性不育突变体小孢子发育的细胞学研究

利用光镜和透射电镜对番茄雄性不育突变体９５３０５的小孢子发育进行了细胞学观察。结果表明,小孢子的败育发生在小孢子母细胞时期、四分体时期以及单小孢子时期。绒毡层细胞行为异常,表现为小孢子母细胞时期提早被破坏解体,或绒毡层细胞多层径向分化,异常肥厚,挤压单核期的小孢子。此外败育方式还有单核中期小孢子无法进行外壁沉积,或小孢子的细胞质被降解成空壳,以及花药畸形,花药上长出许多附属物等异常现象。     

洋葱63A细胞质雄性不育与绒毡层的提早衰退有关

细胞质雄性不育（CMS）机理的细胞学研究在许多作物上已有报道，如拟南芥、大豆、水稻、小麦等，但在国内至今未见关于洋葱的此类报道。利用光镜和透射电镜对洋葱不育系63A和保持系63B进行花药发育过程中显微结构和超微结构观察，结果表明，不育花药花粉母细胞时期，花粉母细胞发育正常，花粉囊形状不规则，绒毡层发育迟缓。四分体时期，四分体形成正常，中层严重退化，绒毡层与药室壁完全脱离，细胞质浓缩、空泡化。利用DNA梯度技术发现不育系绒毡层细胞提早发生程序性死亡。小孢子发育时期，小孢子细胞质发生浓缩、降解，绒毡层完全解体。推测不育系小孢子败育与中层、绒毡层提前衰退有关。     

厚轴茶雄性不育株花药败育的生物学特性和细胞学研究

为明确厚轴茶(Camellia crassocolumnaH. T. Chang)雄性不育株花器发育形态、花药和花粉败育时期及兵细胞学特征,利用体视显微镜、石蜡切片技术、染色体制片和DAPI染色法,对厚轴茶雄性不育株和可育株开花迚程、花器形态、花药发育过程、花粉母细胞减数分裂及小孢子发育过程比较观察。结果显示,厚轴茶花属于完全花,花药其四室、呈蝶形,花药壁发育为基本型,绒毡层细胞其双核,于四分体时期形成分泌型细胞,单核花粉期开始降解,花粉母细胞经过减数分裂Ⅰ、减数分裂Ⅱ和胞质分裂后形成四面体型四分体,小孢子呈三角形,成熟花粉为二细胞型花粉。花蕾发育早期,不育株雄蕊发育正常,与可育株无明显差异。花蕾发育后期,不育株花丝弯曲,花药粘连、干瘪、褐化、坏死,不裂药。不育株减数分裂期绒毡层细胞异常增生、排列混乱,单核至双核花粉期绒毡层延迟降解。不育株花粉母细胞减数分裂过程中存在环状单价体、滞后染色体、染色体桥、染色体缺失、不均等分离、微核和多分体等异常现象。不育株小孢子胞质紊乱,单核期花粉粒相互粘附,花粉壁皱缩变形,花粉细胞质和细胞核模糊不清,成熟花粉细胞空瘪凹陷。研究结果表明,厚轴茶雄性不育花...     

A comparative study on anther development in male-fertile and male-sterile plants of Pelargonium crispumL'Her. ex Ait.

Summary Anther development of male-fertile and male-sterile plants in Pelargonium crispum was anatomically examined. Three cultivars, i.e., Lemon crispum, Crispum minor and Prince Rupert, were used. Lemon crispum and Crispum minor are male-fertile, whereas Prince Rupert is male-sterile. The tapetum in every cultivar examined behaved like an amoeba. The tapetal cells of the anther form plasmodial masses. Then, the plasmodial masses fuse producing a periplasmodium. The periplasmodium degenerates and finally disappears. There are no differences in tapetal behaviour between fertile and sterile anthers. In the sterile anthers the endothecium and lip cells do not develop sufficiently. Young microspores show normal growth at early stages. After the completion of their cell wall formation, however, the microspores in the sterile anthers lose their cytoplasm and become empty. On the other hand, the microspores in the fertile anthers increase the volume of their cytoplasm and become fertile pollen grains.     

新型陆地棉细胞质雄性不育系花器形态学和细胞学观察

 采用石蜡切片法,对一种来源于陆地棉的新型细胞质雄性不育材料的3份转育材料（G22A,H109A和1793A）的花药发育过程进行了花器形态及细胞学观察。花器形态和细胞学观察结果表明,G22A,H109A和1793A三个不育系均花瓣小,花丝短,柱头明显外露,败育彻底,无花粉。败育时期主要发生在造孢细胞增殖时期至小孢子母细胞减数分裂前,表明这3份不育材料不育性状不受细胞核基因的影响。对不育材料与其保持系细胞学观察发现,不育材料与正常花药发育细胞学形态上主要差别是绒毡层发育。不育系绒毡层的着色不明显,大小与中层细胞相似且在整个花药发育过程中不降解,而保持系绒毡层的着色深,在小孢子发育过程中逐渐降解。因此,根据细胞学研究结果可以说明花药中绒毡层发育异常是该来源于陆地棉的新型细胞质雄性不育材料败育的细胞学基础。     

棉花细胞质雄性不育花药的淀粉酶与碳水化合物

棉花细胞质雄性不育花药中淀粉酶同工酶的缺失及其酶活性的降低影响花药中淀粉的水解。不育花药花丝输导组织较差或退化,花粉母细胞对可溶性糖需求的降低使花药中缺乏淀粉的积累。不育花药中缺乏淀粉的积累和淀粉酶同工酶的缺失与花粉母细胞的死亡有关。     

萝卜雄性不育系个体发育过程中3种同工酶活性的比较分析

以3种萝卜雄性不育系及保持系为试材,系统研究了它们在营养生长期及生殖生长期过氧化物酶(POD)同IM、酯酶(EST)同工酶及超氧化物歧化酶(SOD)同工酶活性的变化。结果表明：在营养生长期与生殖生长期的营养器官中,3种不育系的POD活性均低于保持系,而在生殖生长期的花蕾中,不育系的POD活性却高于保持系;EST活性变化表现出与POD相反的趋势;而作为清除氧自由基的SOD与败育关系不大。     

湖北光周期敏感核不育水稻农垦58s花药中的某些生化代谢特点

湖北光周期敏感核不育水稻农垦58s单核早期的不育花药的线粒体呼吸速率和LOX活性分别较可育株低11.9％和5％,而单核晚期相应较可育株低18.4％和17.3％。花药发育从单核期至三核期,可育花药的AsA和GSH含量增高,而不育花药仅分别相当于可育株的35—55％和22—32％,并有脂质氢氧化物积累。随着花药发育,可育花药的AsA-POD,GR和6-GPDH活性逐有增高,至三核期达到最高。而不育花药,随花粉败育,在单核早期至三核期,AsA-POD,GR和6-GPDH活性逐渐降低,至三核期,其活性分别为可育株的26％,22％和19％,不育花药的ME和MDH活性亦较可育株低。IDH活性在单核晚期为可育株的47—80％。细胞还原势低是不育花药特征之一。低的还原势可能导致活性氧代谢失调和花药不育。     

Genetic and cytological analysis of a new spontaneous male sterility in radish (<Emphasis Type="Italic">Raphanus sativus</Emphasis> L.)

A male sterile plant appeared in the radish breeding program at the Hubei Academy of Agricultural Sciences, Hubei, China. In its progeny, a two-type (half of plants male sterile, the other half male fertile) line 01GAB was established. An F₂ population of 260 plants from a cross of male-sterile 01GAB and a male fertile line 9802H segregated for male fertility in a 3:1 ratio indicating that fertility was restored by a single dominant gene, here designated RsMs. A PCR-based DNA marker specific to the male fertility Rfob gene in 9802H was absent in 01GAB. Linkage analysis placed the RsMs locus 10.7 cM away from the Rfo locus. In an F₂ population of hybrids between 01GAB and male fertile 9802B, a co-dominant DNA marker for the RSultr3.2A (a radish sulfate transporter gene) locus was linked to the RsMs locus at 1.5 cM suggesting that fertility restoration in 01GAB was located in the region with known male sterility restorers in radish. However, no maintainer for the 01GAB source of male sterility has been identified so far. Cytological observations have shown that the abnormalities in male sterile anthers first appeared in tapetum at the tetrad stage, followed by a hypertrophy of the tapetal cells at the vacuolate microspore period. These results suggest that male sterility in 01GAB is likely to be genetic in nature, or it may represent a new type of the cytoplasmic male sterility.     

Microsporogenesis and anther wall development in male-sterile and fertile lines of pigeon pea (Cajanus cajan (L.) Millsp.)

Summary Microsporogenesis and anther wall development of male-sterile and fertile lines in pigeon pea (Cajanus cajan (L.) Millsp.) was examined microscopically. Male-sterility was complete and was caused by breakdown at the young tetrad stage. Degeneration of the tapetum by vacuolation occurred during the first division of meiosis and appeared responsible for pollen mother cell breakdown. Sterile plants also differed from the fertile plants in the enlargement of the inner middle layer of the anther wall, and in the lack of development of the endothecium.     

Isozyme Analysis of Some Male Sterile Lines in Sorghum bicolor (L.) Moench

A comparative study of peroxidase and acid phosphatase of anthers was carried out on sorghum to characterize five male steriles and their maímainers. Differences in presence/absence and intensity of bands were observed for the isozyme patterns of all the male steriles and their corresponding maintainers. The better expression of enzymes in sterile anthers emphasized the role they play in breaking down various metabolites that are otherwise important for formation of fertile anthers.     

Relationship Between Phytohormones and Male Sterility in Thermo-Photo-Sensitive Genic Male Sterile (TGMS) Wheat

Summary Fertility alternation normally occurs in a novel type of thermo-photo-sensitive genic male sterile (TGMS) wheat whose recessive genes in the nucleus are expressed as virtually complete sterility under lower temperature and shorter daylight conditions, but as normal fertility under higher temperature and longer daylight conditions during the developmental period from meiosis of pollen mother cells (PMCs) to the early mononucleate stage of microsporogenesis. In order to provide more information about the mechanism(s) of fertility alternation in TGMS wheat, the relationship between fertility alternation and levels of phytohormones was analyzed by endogenous hormone quantification and exogenous hormone treatments in the present research. Sown at an earlier date with a relatively low temperature and short daylight, TGMS wheat was male sterile phenotypically. Pollen abortion mainly occurred at the mononucleate to bi-nucleate stages of microsporogenesis and pollen abortion was mainly of the spherical type. Levels of endogenous indole-3-acetic acid (IAA), abscisic acid (ABA) and gibberellic acid (GA₃) in the anthers were at lower levels, and zeatin riboside (ZR) was higher. Sown at a later date, when the temperature during the critical period for fertility alternation was relatively high and the daylight was long, TGMS wheat was almost as fertile as the normal genotype and the contents of endogenous IAA, ABA and GA₃ in anthers were at higher levels, and the content of ZR was at a lower level. At the critical period for fertility alternation, treatments with exogenous IAA, ABA, GA₃ or aminoethoxy vinylglycine (AVG), an inhibitor of ethylene biosynthesis, decreased the frequency of sterile pollen grains and increased the frequency of self-pollinated fertile florets. Treatment with exogenous 2-chloroethyl phosphonic acid (CEPA), a releaser of ethylene, increased the frequency of sterile pollen grains and decreased the frequency of self-pollinated fertile florets. It was thus suggested that the fertility alternation of TGMS wheat was closely related to levels of phytohormones in the anthers, and changes of endogenous hormone levels were among the important factors responsible for the fertility alternation of TGMS wheat.     

黏类小麦细胞质雄性不育相关基因cMDH的克隆与表达分析

为深入研究黏类小麦雄性不育的分子遗传机制,利用抑制差减杂交技术构建了黏类小麦育性相关基因的二核期SSH文库.经文库筛选后,得到一个在可育文库中表达的与胞质苹果酸脱氢酶基因同源的EST序列.以该EST序列为信息探针,经电子克隆、RT-PCR、PCR克隆与序列分析,获得了小麦胞质苹果酸脱氢酶(cytosolic malate dehydrogenases,cMDH)基因的cDNA与DNA序列,利用荧光定量PCR技术对该基因在不育株和可育株花药中的表达进行了分析,并比较了MDH在小麦不育株和可育株中的活性变化.结果表明,该基因的cDNA序列长1213 bp,编码333个氨基酸;DNA序列长2 908 bp,含有7个外显子和6个内含子;该基因在不育株和可育株花药发育3个时期(单核、二核和三核)的表达均表现为先升后降的模式,而且该基因在不育株花药发育的二核期和三核期的表达相对于可育株被明显抑制;MDH在小麦不育株和可育株中的活性变化趋势与定量结果一致.推测该基因在花粉发育早期表达,它的下调表达可能影响了小麦雄蕊发育过程中的能量供应而导致雄性不育.     

甘蓝型油菜光、温敏雄性不育系Huiyou50S花粉败育的细胞学观察

Huiyou50S是从甘蓝型油菜品种汇油50中发现的半不育株选育而成的光、温敏雄性不育系,育性受隐性核基因控制,在高温、长光周期下不育而在低温、短光周期条件下可育。本文通过半薄树脂切片、扫描电镜、花粉压片染色、花药整体透明方法对Huiyou50S及其近等基因系Huiyou50F的花药发育过程及花粉形态观察比较。结果表明, Huiyou50S的花药造孢细胞、花粉母细胞、减数分裂、四分体阶段均正常,在单核期出现明显异常,虽然能形成花粉壁,但小孢子细胞质收缩、解体,最终只剩余空瘪的花粉壳; Huiyou50S的花药绒毡层在单核后期提前降解,绒毡层解体速度快于可育株Huiyou50F。Huiyou50S的小孢子的完全败育主要发生在单核时期,绒毡层发育与小孢子异常有某种关联,该结论为油菜光、温敏雄性不育类型划分及育种应用提供了依据。     

芝麻核雄性不育系ms86-1小孢子败育过程的超微结构

运用透射电子显微镜对芝麻核雄性不育系ms86-1的可育和不育花药进行了超微结构的比较观察。根据小孢子的细胞学形态特征,将芝麻花粉发育过程划分为小孢子母细胞形成期、减数分裂期、四分体期、单核小孢子早期、单核小孢子中期、单核小孢子晚期、花粉成熟期7个时期。对比观察表明芝麻核雄性不育的败育迹象起始于小孢子母细胞形成期,并伴随着进一步发育,败育现象逐渐明显,小孢子母细胞形成期小孢子母细胞壁形状不规则;减数分裂期小孢子母细胞壁严重扭曲变形,质膜外缺少早期外壁成分--原基粒棒;四分体期胼胝质壁外沉积物异常,呈绒毛状;四分体解体后形成畸形小孢子,孢子外壁不健全,绒毡层异常肥厚、降解延迟,释放极少量的畸形乌氏体;随后小孢子愈发皱缩,胞质凝集,内含物减少并逐渐凝聚成一团电子致密物质,最终走向完全败育。本研究揭示了不育小孢子的败育过程和败育特征,为深入研究芝麻核雄性不育败育机理奠定了基础。     

Genetic investigation and cytological comparison of two genic male sterile lines 9012A and MSL in <Emphasis Type="Italic">Brassica napus</Emphasis> L.

Genic male sterility (GMS) is one of the effective pollination control systems in hybrid rapeseed (Brassica napus L.) breeding. The recessive GMS lines 9012A and Lembke (MSL) have made a great contribution to the utilization of heterosis in rapeseed. Cytological observation and genetic investigation were employed to reveal anther abortion characters and genetic relationship of 9012A and MSL. Cytological and transmission electron microscopy observation revealed that anther abortion of 9012A and MSL initiated at the microsporocyte stage, both lines showed premature or retarded degradation of tapetum. Some anthers in 9012A developed to the microspore stage since their tapetal cells changed to the secretory type. However, MSL anthers did not progress to the tetrad stage, since the tapetum did not change to the secretory type. Genetic investigation revealed that MSL and 9012A had the same temporary maintainer system, while their cytoplasm types were different. The cytoplasm of MSL was similar to Cam while 9012A was similar to Nap type. The results laid a solid foundation for utilization of the two male sterile lines in rapeseed hybrid breeding.