甘蓝型油菜隐性上位互作核不育材料1665花药败育的细胞学研究

采用石蜡切片方法,对甘蓝型油菜隐性上位互作核不育材料1665的可育株与不育株花药进行细胞学观察.结果显示:(1)不育株花药在花粉母细胞减数分裂时期出现异常,部分花粉母细胞细胞分裂相不均等分裂或分裂异常.导致部分四分体形状异常.(2)不育株绒毡层细胞在四分体时期开始生长膨大,单核花粉时期出现液泡化和巨型化,侵占药室,使得小孢子不能正常释放或无法继续发育;部分释放出的小孢子未及时形成花粉壁,阻碍花粉继续发育.不能发育形成二核期和三核期花粉,导致花药败育.     

新型甘蓝型油菜温敏核不育系SP2S的花药发育解剖学观察

SP2S是西北农林科技大学选育的甘蓝型油菜温敏核不育系,本文采用半薄树脂切片、扫描电镜对SP2S及其可育近等基因系SP2F的花药发育及花粉形态进行观察比较,发现SP2S花药发育在减数分裂时期出现异常,单核花粉时期彻底败育。其主要特征是：减数分裂时期绒毡层已经径向肥大且出现大液泡,胼胝质不能及时降解,使得单核小孢子相互粘连在一起,小孢子无花粉壁的形成且细胞质物质逐渐降解,最后小孢子仅剩下空壳残留物,聚集在一起。SP2S败育特征与现有的核不育材料不同,表明其有可能是一种新型温敏核不育材料。     

温敏核不育小麦可育花药和败育花药发育观察

对温敏核不育小麦百农不育系（Bainong sterility,BNS）的可育和不育花药结构进行对比观察。在减数分裂期、小孢子早期和小孢子晚期,可育花药与不育花药的结构相同。小孢子分裂形成二胞花粉后,可育花粉中随着大液泡的分解,细胞质内含物增加,其中出现一些颗粒状物质。不育花药中,小孢子也可分裂形成二胞花粉,但营养细胞的大液泡不分解,细胞质也不增加,最终花粉中的细胞质消失,花粉败育。该种温敏核不育小麦的花粉败育时间发生在二胞花粉早期,可能和其大液泡没有适时分解有关。花粉败育时间的确定为进一步深入研究该种雄性不育小麦的败育机制打下了基础。     

温敏雄性核不育水稻可育花药与不育花药的钙分布

用焦锑酸盐沉淀法研究了温敏雄性核不育水稻在减数分裂时期和单核早期可育花药与不育花药的钙分布.结果表明：在减数分裂时期,可育花药小孢子母细胞和药室内的钙颗粒很少,而不育花药小孢子母细胞中分布许多的钙颗粒,特别是药室中的钙颗粒异常丰富,小孢子母细胞减数分裂异常,细胞质收缩退化.在单核早期,可育花药花粉内的钙颗粒极少,花粉表面分布许多钙颗粒,而不育花药花粉内分布许多钙颗粒,药室内的钙颗粒仍然非常丰富.可育花药维管束鞘细胞体积大且形状规则,细胞内的钙颗粒很少,而不育花药维管束鞘细胞体积小且形状不规则,细胞内的钙颗粒较多.     

白菜核雄性不育两用系可育和不育花药ATPase的分布

对白菜核雄性不育两用系可育花药和不育花药的ATPase做了定位分析。可育花药的花粉母细胞核中积累了大量的ATPase反应颗粒,而细胞质中ATPase反应颗粒较少,但在线粒体中特异地聚集了一些大的ATPase反应颗粒。减数分裂后,小孢子细胞质中ATPase反应颗粒明显增加。随着小孢子发育,其细胞质中ATPase反应颗粒逐渐减少,但在线粒体中又特异性地聚集了较多的AT-Pase反应颗粒。当花药发育到二胞花粉时期,花粉和绒毡层细胞中的ATPase反应颗粒已很少了。不育花药的花粉母细胞中呈现较多的ATPase,然而在线粒体中很少。异常四分体小孢子细胞质中虽然有较多的ATPase反应颗粒,但还是通过细胞质收缩和质壁分离方式退化。对可育花药的花粉母细胞线粒体中特异出现的簇状ATPase分布现象进行了分析,讨论了不育花药中花粉母细胞线粒体ATPase的异常与花粉败育的可能关系。     

太谷核不育小麦花药内游离脯氨酸和总氨基酸含量的变化及其与育性的关系

本文对显性单基因控制的太谷核不育小麦不同发育阶段的可育株和不育株的花药及雌蕊内游离肺氨酸和游离总氨基酸的含量进行了分析。结果表明:(1)在小孢子母细胞减数分裂期,不同育性花药之间游离脯氨酸的含量无明显差异,且含量较低。(2)在小孢子单核初期,可育花药内游离脯氨酸的含量显著高于不育花药,是不育花药的7倍,比减数分裂期增加20倍,高达其干重的1.65％,占其游离总氨基酸的50％。(3)在雌蕊中,游离脯氨酸的含量远远低于花药,不同育性植株之间差异不很明显。(4)关于游离总氨基酸的含量,在花药中减数分裂期,不同育性植株之间无明显差异;在小孢子单核初期,可育株高于不育株。在雌蕊中,相应于小孢子单核初期时,可育株稍高于不育株,受精后迅速趋于一致,但整个变化幅度不大。     

小麦粘类雄性不育系生化标记及小孢子细胞色素氧化酶同工酶研究

对4种同核异质小麦粘类非1BL/1RS雄性不育系、保持系和恢复系的幼苗叶片、乳熟期籽粒以及不育系、恢复系和F1小孢子发育四分体至三核期花药进行了细胞色素氧化酶（COD）同工酶聚丙烯酰胺凝腔电泳（PAGE）分析。结果表明：（1）幼苗叶片COD同工酶谱带可以标记4种不育系和保持系;乳熟期籽粒COD同工酶谱带可以将4种不育系、保持系及恢复系区别开。（2）COD在不育系小孢子败育时或败育之前（单核到二核期）酶量降低,面在三核期酶量升高。（3）相同胞质背景下引入不同核恢复基因或不同胞质背景下引入桢核恢复基因,F1小孢子COD同工酶谱带之间有差异。可以将不同发育时期COD同工酶谱带作为鉴别1种不育系以及不育系、保持系、恢复系（“三系”的可靠生化标记）。     

灯盏花雄性不育种质鉴定与花药发育的细胞学研究

对云南泸西栽培灯盏花群体进行调查,发现了灯盏花雄性不育种质个体,其出现频率约为1.06×10-4.对所发现的灯盏花不育株形态特征及其花药发育过程进行了观察,并对花粉活力进行鉴定.结果显示:(1)灯盏花不育株根、茎、叶形态与正常可育植株基本相似,管状花小,花丝短,花药瘦小,无花粉粒散出或花粉无活力.(2)灯盏花在其花药发育的小孢子母细胞时期、四分体时期、小孢子时期和单核早期,由于绒毡层细胞液泡化、提前解体,不能为小孢子或花粉发育提供所需物质,导致小孢子母细胞和四分体解体,产生无花粉的花药;或小孢子和单核花粉胞内降解,形成不同形状和外壁纹饰的败育花粉.研究认为,灯盏花花药绒毡层异常是其花粉败育的主要原因.     

甘蓝型油菜几个雄性不育系花药发育的细胞形态学研究

选用甘蓝型油菜6个细胞质雄性不育系和1个细胞核雄性不育系为材料,与可育系比较,确定花药发育受阻的时期和方式。根据研究结果,将其雄性不育系分为三类:1.湘矮A,Po-aA,陕2A和7s-3A花药发育受阻于孢原细胞分化期,没有分化形成花粉囊。2.萝AⅠ和萝AⅡ花药发育受阻于四分体至单核花粉期。败育方式为小孢子难以从四分体中释放出来,或释放出来后细胞质液泡化,核不能分裂,花粉壁发育不良。此外,还见到绒毡层径向肥大、延迟消失和维管束分化不良等异常现象。3.宜3A为核不育系,花药发育受阻于花粉母细胞期。败育方式为花粉母细胞死亡,减数分裂异常,或不能进行减数分裂。绒毡层和维管束一般都能正常发育。     

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     

红菜薹雄性不育系花药败育的细胞形态学观察

采用石蜡切片技术,在光学显微镜下系统研究了红菜薹(Brassica campestris L.ssp．chinensis L.var．utilis TsenetLee．)波里马胞质雄性不育系(Polima CMS)、红菜薹萝卜胞质雄性不育系(Ogura CMS)及相应保持系花药发育过程的细胞形态学特征。观察结果表明：红菜薹Polima CMS花药发育受阻于孢原细胞阶段,不形成花粉,属无花粉型,此不育系花药不形成绒毡层和中层;而红菜薹Ogura CMS花药败育发生于小孢子母细胞期或四分体时期,表现为绒毡层细胞异常,挤压四分体,导致四分体和绒毡层同时解体而败育。     

玉米黄早4雄性不育系、保持系过氧化物酶同工酶的比较研究

在玉米黄早4雄性不育系、保持系的10个组织(叶、根、茎、芽鞘、胚轴、苞叶、穗轴、花丝、雌蕊、花药)中共检测出18种正、负极向过氧化物酶,其中有7个组织不育系与保持系间过氧化物酶没有差异,只有在3个组织中(叶、茎、花药)不育系与保持系间的过氧化物酶存在差异,说明玉米黄早4雄性不育系及保持系的过氧化物酶可能由细胞核基因编码。不育系与保持系个别组织内过氧化物酶存在差异,可能是由于核内编码过氧化物酶的基因表达异常所引起,而这种表达异常,可能是与不育系中不育细胞质基因调控核基因的表达有关。     

Microsporogenesis and tapetal development in fertile and cytoplasmic male-sterile sugar beet (Beta vulgaris L.)

Summary The development of sporogenous and tapetal cells in the anthers of male-fertile and cytoplasmic male-sterile sugar beet (Beta vulgaris L.) plants was studied using light and transmission electron microscopy. In general, male-sterile anthers showed a much greater variability in developmental pattern than male-fertile anthers. The earliest deviation from normal anther development was observed to occur in sterile anthers at meiotic early prophase: there was a degeneration or irregular proliferation of the tapetal cells. Other early aberrant events were the occurrence of numerous small vesicles in the microspore mother cells (MMC) and a disorganized chromatin condensation. Deviations that occurred in sterile anthers at later developmental stages included: (1) less distinct inner structures in the mitochondria of both MMC and tapetal cells from middle prophase onwards. (2) dilated ER and nuclear membranes at MMC prophase, in some cases associated with the formation of protein bodies. (3) breakdown of cell walls in MMCs and tapetal cells at late meiotic prophase. (4) no massive increase in tapetal ER at the tetrad stage. (5) a general dissolution of membranes, first in the MMC, then in the tapetum. (6) abortion of microspores and the occurrence of a plasmodial tapetum in anthers reaching the microspore stage. (7) no distinct degeneration of tapetal cells after microspore formation. Thus, it seems that the factors that lead to abortive microsporogenesis are structurally expressed at widely different times during anther development. Aberrant patterns are not restricted to the tetrad stage but occur at early prophase.     

A Comparative Study of Anther and Pollen Development in Male Fertile and Male. Sterile Green Onion (Allium fistulosum L.)

Anther and pollen development in male-fertile and male-sterile green onions was studied. In the male-fertile line, both meiotic microspore mother ceils and tetrads have a callose wall. Mature pollen grains are 2-celled. The elongated generative cell with two bended ends displays a PAS positive cell wall. The tapetum has the character of both secretory and invasive types. From microspore stage onwards, many oil bodies or masses accumulate in the cytoplasm of the tapetal cells. The tapetum degenerates at middle 2-celled pollen stage. In male-sterile line, meiosis in microspore mother cells proceeds normally to form the tetrads. Pollen abortion occurs at microspore with vacuole stage. Two types of pollen abortion were observed. In type I, the protoplasts of the microspores contract and gradually disintegrate. At the same time the cytoplasm of microspores accumulates oil bodies which remain in the empty pollen. The tapetal cells behave normally up to the microspore stage and early stage of microspore abortion, but contain fewer oil bodies or masses than those in the male-fertilt line. At late stage of microspore abortion, three forms of the tapetal ceils can be observed: (1) the tapetal cells with degenerating protoplasts become flattened, (2) the tapetal cells enlarge but protoplasts retractor, (3) the cells break down and tile middle layer enlarges. In type Ⅱ, the cytoplasm degenerates earlier than the nucleus of the microspores and no protoplast is found in the anther locule. There are fibrous thickenings iii the endothecium of both types. It is difficult to verify whether the tapetum behavior and pollen abortion is the cause or the effect.     

MALE STERILITY IN SOYBEAN (GLYCINE MAX). I. PHENOTYPIC EXPRESSION OF THE ms2 MUTANT

The effects of a nuclear male-sterile mutant (ms2) of soybean, Glycine max (L.) Merr., on anther development were analyzed by means of light- and electron-microscopy. The structure of microspore mother cells (MMCs) in male-sterile plants was identical to that of male-fertile plants. Meiosis was completed, and tetrads of microspores formed. Microspores degenerated after the deposition of primexine and probacullae. The sheath of callose surrounding microspores did not dissolve. No structural abnormalities of the microspores were detected before the onset of degeneration. The tapetal and anther wall layers were characterized by aberrant development. Tapetal abnormalities included premature vacuolation, a persistent inner tangential cell wall, failure to differentiate normal concentrations of endoplasmic reticulum and dictyosomes, disruption of plastids, and premature degeneration. Malfunction of the tapetal layer preceded, and may have induced, microspore degeneration. Gross anther morphology was not influenced until advanced stages of development.     

Tapetum and middle layer control male fertility in Actinidia deliciosa

Background and Aims

Dioecism characterizes many crop species of economic value, including kiwifruit (Actinidia deliciosa). Kiwifruit male sterility occurs at the microspore stage. The cell walls of the microspores and the pollen of the male-sterile and male-fertile flowers, respectively, differ in glucose and galactose levels. In numerous plants, pollen formation involves normal functioning and degeneration timing of the tapetum, with calcium and carbohydrates provided by the tapetum essential for male fertility. The aim of this study was to determine whether the anther wall controls male fertility in kiwifruit, providing calcium and carbohydrates to the microspores.

Methods

The events occurring in the anther wall and microspores of male-fertile and male-sterile anthers were investigated by analyses of light microscopy, epifluorescence, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL assay) and transmission electron microscopy coupled with electron spectroscopy. The possibility that male sterility was related to anther tissue malfunctioning with regard to calcium/glucose/galactose provision to the microspores was also investigated by in vitro anther culture.

Key Results

Both tapetum and the middle layer showed secretory activity and both degenerated by programmed cell death (PCD), but PCD was later in male-sterile than in male-fertile anthers. Calcium accumulated in cell walls of the middle layer and tapetum and in the exine of microspores and pollen, reaching higher levels in anther wall tissues and dead microspores of male-sterile anthers. A specific supply of glucose and calcium induced normal pollen formation in in vitro-cultured anthers of the male-sterile genotype.

Conclusions

The results show that male sterility in kiwifruit is induced by anther wall tissues through prolonged secretory activity caused by a delay in PCD, in the middle layer in particular. In vitro culture results support the sporophytic control of male fertility in kiwifruit and open the way to applications to overcome dioecism and optimize kiwifruit production.     

粳稻细胞质雄性不育系、保持系POD同工酶的比较研究

粳稻３４１Ａ、３４１Ｂ的不同器官和花药及雌蕊的不同发育时期共检测出９种ＰＯＤ同工酶谱带,其中ＰＯＤ＿５、ＰＯＤ＿６在所有１０个器官中存在,因此可以认为是粳稻３４１Ａ、３４１Ｂ的基本谱带。ＰＯＤ＿８、ＰＯＤ＿９谱带仅在少数器官中存在,具有一定的器官专一性。花粉粒发育的两个关键时刻,ＰＯＤ同工酶均多于保持系,这可能是雄性不育细胞质基因对核基因表达的调控作用所致,细胞质基因与细胞核基因存在着一定的相互关系,雄性不育可能是这两种基因共同作用的结果。     

普通小麦T型和V型雄性不育系同工酶的比较研究

对不同发育时期花药的COD和POD同工酶电泳分析表明,T型不育系与其保持系、V型不育系与可育株以及两不育类型之间在COD同工酶图谱和酶活性方面的差异主要表现在单核期;而POD同工酶的差异则是在单核早期。     

Studies on the Free Amino Acid Content in Anthers of Malesterile and Fertile Plants of Taigu Wheat,with Special Reference to Free Proline Content

Biochemical analyses were made on anthers and pistils at various developmental stages of both male-sterne and fertile plants of Taigu wheat. Analyses ineluded total free amino aeids and free proline. The following results were obtained: 1. There was no significant difference between the content of free proline in anthers of male-sterile and fertile plants at reduction division of mierospore mother cells. 2. In anthers with early uninucleate miorospores, the content of free proline of fertile plants was remarkably higher than that of male-sterile plants. It is interesting to note that at this stage the content of free proline in fertile plants rose to 1.65% of the dry weight of the anther, constituting 50% of the total free amino acids, and amounted to 7-fold of that in male-sterile plants. This result is in line with the results obtained with most cytoplasmic malesterile plants reported by other workers, although malesterility in Taigu wheat is controlled by the nueleus. 3. In pistils, at the stages eorresponding to the early uninneleate and the binueleate stages of the pollen, the free proline content of fertile plants was twice as much as that of the male-sterile plants. This differenee disappeared gradually after fertilization. 4. Tile content of total free amino aoids did not fluetuate as much as the free proline content. There was no differenee in anthers of both types of plants during reduction division of mierospore mother cells. In anthers with early uninueleate pollen grains, total free amino acid content of fertile plants exceeded that of male-sterile plant, the difference levelled off at latter stages. In pistils, before fertilization, the content of total free amino acids of the fertile phmts was slightly higher than that of the male-sterile plants. After fertilization t}fis difference was no nmre noticeable.