温光敏核不育水稻N28S无花粉败育的显微结构观察

采用石蜡切片和荧光显微技术观察了温光敏核不育水稻N28S无花粉败育过程中的显微结构变化,结果显示：N28S的小孢子母细胞形成后细胞质变得稀薄,一部分不能进行减数分裂,一部分减数分裂阻滞在细线期或胞质分裂异常,最终所有细胞液泡化解体消失。在此过程中,还观察到小孢子母细胞在细线期胼胝质壁不产生或提早消失,以及小孢子发育后期花药壁绒毡层的异常解体。认为N28S的无花粉败育是由小孢子母细胞的细胞质异常引起的,胼胝质壁和绒毡层的异常是结果而不是原因。     

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     

辣椒雄性不育系与可育系小孢子发生的细胞学观察

为了探讨辣椒雄性不育花药败育时期和方式,以辣椒雄性不育系1442A、13733A及其可育系为试材,进行了研究。结果发现：败育现象从造孢细胞时期以后每个阶段都有发生,败育形式有造孢细胞液泡化、畸形、拉长、细胞间隙大;绒毡层细胞径向过度伸长,高度液泡化,且出现多层细胞,严重挤压小孢子母细胞,解体较晚且充塞花粉囊室;薄壁细胞取代了药室内壁、中层、绒毡层和小孢子母细胞的分化;药室内壁、中层层数增加,绒毡层细胞肥大,造孢细胞或花粉母细胞分解解体;由于花粉母细胞胼胝质壁不降解而无法释放出四分体小孢子;染色浅、细胞质被降解成空壳的单核期小孢子因缺乏营养物质而败育。     

橡胶树花药和小孢子发生与发育过程的观察

应用石蜡切片法．观察橡胶树的实生树和RRIM600、GT-1品系的花药壁以及小孢子的发生和发育过程,得到如下结果：1．实生树的花药壁通常由四层细胞组成,发育形式为双子叶型。药室内壁细胞在发育后期进行径向条纹加厚．至花药开裂时仍保留着原生质体。中层由一层或不规则的两层细胞组成,在小孢子单核期消失。绒毡层细胞具单棱或双核,属分泌型,至花粉发育到三细胞时消失。小孢子母细胞减数分裂为同时型。成熟花粉粒具三十细胞。精细胞椭圆形,在光镜下不能区分出细胞质鞘和核仁。所观察的实生树雄花,多数发育正常,很少有空秕的花粉。2．RRIMB00品系的花药和小孢子发生与发育和实生树相似,但至后期只有少数花粉发育正常,多数成为大小不等的败育花粉;此外也有一些败育的雄花。3．GT-1的花药在小孢子母细胞减数分裂时,绒毡层细胞的体积开始异常增大并液泡化．小孢子在四分体内解体或分离后成为空秕花粉。     

雄性可育与雄性不育籽粒苋小孢子发育研究

栽培种籽料苋（lmaranthus hypochondriacus L。）是一种很有潜力的新型作物。它营养价值高、蛋白质含量丰富、氨基酸平衡好、耐旱、耐盐碱和酸、抗逆性强、适应性广,被认为易极有潜力的、为全球提供粮食的替代作物之一。但是籽粒苋于粒重仅0．7－1．2g,种子易散落。于是,和许多其它植物一样,籽粒苋中也找到了雄性不育株。但是它的小孢子发育过程及其败育时期和不育特征尚不清楚,为它的杂交育种研究带来不便。本文通过电镜对雄性可育和不育的两种籽粒苋小孢子进行观察。发现不育小孢子败育起始于四分体释放以后的单核花粉期。在此之前小孢子的发育是一样的。花粉分化早期,孢原组织分化出初级造孢组织、绒毡层、中间层、药壁内层和表皮层（图1）;造孢组织继续分裂,细胞不断扩大,形成小孢子母细胞（图2）;小孢子母细胞不断增大,周围积累胼胝质并逐渐与绒毡层分离,出现大液泡（图3）;小孢子母细胞减数分裂,四分体形成,包埋于胼胝质中;绒毡层有丝分裂,有双核细胞;大液泡消失;细胞壁开始降解（图4）。胼胝质逐渐消失,小孢子从四分体中释放以后（单核花粉期）,在雄性可育籽粒苋里,小孢子有丝分裂、迅速膨大变圆,可见两个深色雄仁,花粉壁加厚（图5）;进入收缩期,绒毡层降解,突入花药腔,环绕小孢子周围（图6）;花粉壁不断加厚,小孢子更趋成熟（图7）,直直形成内含大量淀粉的完全成熟花粉粒（图8）。而在雄性不育籽粒苋里,出现如下不育特征：小孢子粉壁未能进一步加厚,小孢子形状变得怪异（图13）;花粉内含物溶解,空泡化,成为不育花粉（图14）。小孢子在花药中的发育完全依赖绒毡层细胞提供所需的营养物质和信息,绒毡层异常必然导致花粉败育,胼胝质降解不影响小孢子母细胞减数分裂,而是影响小孢子初生外壁的发育,从而导致小孢子发育退化。籽粒苋花粉败育过程中未见胼胝质降解,其原因有待进一步研究。有报道,正常花粉发育过程中常含有大量液泡,籽粒苋可育花粉的发育过程也证实液泡的发育与花粉粒的充实、花粉的形状有密切关系,而不育花粉中小液泡逐渐膨大,形成空泡后破裂。     

无花粉型水稻温敏核不育系籼S的育性表现与细胞学观察

温敏核不育水稻籼S是从优质常规稻籼黄占自然突变而来的一个无花粉型光温敏核不育种质资源。在广州(23°08′N)自然条件下,一年中具有明显的“可育-不育-可育”的育性转换,5月初至10月底为稳定不育期。在人控光温条件下,低温诱导其由不育转为可育需要较长的持续时间,日均温21℃需7d以上,23.5℃需15d以上。细胞学观察表明其无花粉败育主要是由减数分裂时期的异常引起的,表现为小孢子母细胞粘连与液泡化、减数分裂受阻于前期Ⅰ的细线期、进行无丝分裂与异常的胞质分裂,始终没有正常四分体的形成,而是产生大小不同、核数不等的异常细胞,并最终解体消失。其花粉败育特点不同于以往研究过的光温敏核不育水稻,具有花粉败育时期早而败育彻底的特点。     

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。     

芡实花粉败育的细胞学观察

芡实（ＥｕｒｙａｌｅｆｅｒｏｘＳａｌｉｓｂ．）花粉的败育主要发生在小孢子母细胞减数分裂时期和小孢子单核期与二核期。形成的不正常四分体中有一些具有不完整的横隔壁,一些则完全不形成横隔壁,而发育成具４个核的原生质团;气候异常是造成该时期不育现象的主要原因之一。小孢子单核期及二核期存在发育异常的孢子。绒毡层的提前发育、过早解体、肥大生长及延迟退化是花粉败育的一个重要原因。     

冬季低温条件下香石竹小孢子败育的细胞学研究

利用压片法及石蜡切片法观察冬季低温下香石竹小孢子发育过程,以明确低温导致香石竹小孢子败育的因素,为杂交育种奠定基础。结果表明:(1)冬季低温下香石竹只有部分小孢子发育正常,经过小孢子母细胞、减数分裂和四分体等时期,最后发育成花粉。(2)石蜡切片法观察到冬季低温下香石竹1.5~1.6cm长花蕾中有61%的花粉母细胞发生败育,1.7~1.8cm长花蕾中有71%的花粉母细胞发生败育。(3)部分已经进入四分体时期的小孢子胼胝质未能及时溶解,妨碍了小孢子释放而导致败育。研究认为,花粉母细胞和四分体的发育异常是冬季低温下香石竹小孢子败育的主要原因。     

甜菜无融合生殖单体附加系M14花粉发生与发育的超微结构研究

运用透射电子显微镜技术,对甜菜无融合生殖单体附加系M14的小孢子发生、雄配子体发育以及相应的花药壁发育过程进行超微结构的观察研究,以阐明甜菜无融合生殖单体附加系M14花粉发生与发育超微结构特点以及花粉败育的时期和败育的细胞学特征.结果显示:(1)小孢子母细胞减数分裂正常,分裂期间细胞质具有明显的"细胞质改组"现象,主要表现在核糖体减少,质体、线粒体的结构发生规律性的变化,有利于孢子体向配子体的转变.M14减数分裂的胞质分裂为同时型,前期Ⅱ和中期Ⅱ形成"细胞器带";正常发育的花粉,小孢子分裂形成营养细胞和生殖细胞;生殖细胞脱离花粉壁,生殖细胞游离于营养细胞的细胞质中,最初具细胞壁,而后消失,且生殖细胞壁成分与花粉内壁成分相似.(2)三细胞型的成熟花粉含有一个营养细胞和两个具有尾突的精子;每个精子通过两层质膜与营养细胞隔开,含有一个大的精核,长尾突内含少量的细胞质以及纤丝状结构.(3)生殖细胞和精子中缺乏质体.(4)花粉的败育起始于小孢子,大部分受阻于单核-二细胞花粉期,其败育特征为花粉内液泡吞噬作用导致细胞器解体,绒毡层细胞过早解体或肥大生长致使营养供应受阻,可能是导致单核-二细胞花粉败育的主要细胞学原因.研究表明,白花甜菜第九号染色体的附加可能是导致M14大量花粉败育的遗传学因素.     

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.     

大果白刺雄性不育系的细胞学研究

利用石蜡切片法对大果白刺花药进行细胞学研究,探讨雄性不育系发生败育的时期和方式以及雄性败育与药壁组织间的关系.结果表明:不育系小孢子母细胞形成前期,花药各部分结构发育正常.随着绒毡层的异常解体,多种异常现象相继出现,包括小孢子母细胞液泡化,中层、药室内壁、药隔细胞液泡化,细胞畸形,药壁细胞非正常解体等.退化后整个花药萎缩干瘪,不能开裂,无花粉.因此,大果白刺雄性不育系的绒毡层生理异常并提前退化是导致雄性不育的主要原因.     

Distribution of cellulosic wall in the anthers of Arabidopsis during microsporogenesis

Key message

Cellulose-specific staining revealed that tapetal cells and microsporocytes lose cellulosic walls before the onset of meiosis. Cellulosic wall degradation in microsporocytes might be independent of tapetal cells (or TPD1).

Abstract

Some cell types in a variety of angiosperms have been reported to lack cell walls. Here, we report that the tapetal cells of the anther of Arabidopsis thaliana did not appear to have a cellulosic wall based on staining with Calcofluor and Renaissance 2200. During sporogenous cell formation, cellulosic wall was present in all anther tissues. However, before meiosis it was almost absent on the tapetal cells and on the microsporocytes. In a sporocyteless/nozzle (spl/nzz) mutant, which lacks several components (microsporocytes, tapetum, middle layer and endothecium), cellulosic wall was detected in all anther cells. In another mutant, tapetum determinant1 (tpd1), which lacks tapetum and has more microsporocytes, cellulosic wall was almost absent on the microsporocytes before meiosis, similar to the wild type. These results suggest that the tapetum cells and microsporocytes lose cellulosic walls during microsporocyte formation, and that cell wall degradation occurs downstream of SPL/NZZ and is independent of TPD1.     

掌叶大黄花药的发育和异常

观察了掌叶大黄花药的发育过程及异常现象,主要结果为：花药四室,药壁发育属单子叶型,腺质绒毡层。小抱子母细胞的减数分裂为同时型,四分体为正四面体型。从小孢子母细胞减数分裂开始到四分体时期,规律性沉积胼胝质。成熟花粉为三细胞。减数分裂过程中还见到单个或多数染色体游散于赤道板外,落后染色体、染色体桥和微核等异常,平均变异率6．29％。     

Regulation of β-1,3-glucanase activity in developing anthers of Lilium

Lilium microsporocytes have specialized walls consisting of callose, presumably a polymer of β-1,3-glucose. At the termination of meiosis, the walls undergo sudden and rapid dissolution resulting in the liberation of young microspores. This event is correlated with a sharp peak in activity of β-1,3-glucanase. The activity is localized in the somatic region of the anther with less than 7.3% of the total activity associated with the microsporocytes at any time during meiosis. Thus β-1,3-glucanase appears to be an enzyme necessary to the development of meiotic cells but whose action is mediated by the surrounding somatic tissue rather than by the meiotic cells themselves.     

Cellular organisation in meiotic and early post-meiotic rice anthers

We have used fluorescent, confocal laser and transmission electron microscopy (TEM) to examine cellular organisations, including callose (1,3-beta-glucan) behaviour, in meiotic and early post-meiotic rice anthers. These features are critical for pollen formation and provide information to better understand pollen sterility caused by abiotic stress in rice and other monocotyledonous species. Among organelles during meiosis, abundant plastids, mitochondria and nuclei of the anther cells show distinctive features. Chloroplasts in the endothecium store starch and indicate a potential for photosynthetic activity. During meiosis, the middle layer cells are markedly compressed and at the tetrad stage are either vacuolated or filled with degenerating electron-opaque organelles. Viable mitochondria, stained with Rhodamine 123, are seen in the endothecium and tapetum, but the mitochondria in the middle layer are not stained during meiosis. The radial walls of the tapetum are disorganised and degenerating, indicating the formation of a syncytium; pro-orbicules are located at the locular walls at the tetrad stage. Immunohistochemical studies show that the sporogenous cells are entirely enveloped by a thick callosic layer at early meiosis. Cell plate callose was assembled in a plane between the dyad cells. In the tetrads, however, callose formed only at the centre, showing that the tetrad microspores are not enveloped but separated by callose walls. Thick, undulating electron-opaque walls around the tetrads indicate the beginning of exinous microspore wall differentiation.     

Sporophyte-gametophyte interactions between anther and male gametophyte in petunia

Sporophyte-gametophyte interactions between anther and male gametophyte were investigated in two (fertile and sterile) clones of petunia (Petunia hybrida L.) with different reproductive strategies. Structural and functional reorganization of sporophyte tissues in the developing anther of fertile clone is closely coordinated with each of the successive stages of male gametophyte development (from meiosis to the formation of binuclear pollen) and comprises not only destruction of tapetum and three middle layers of the wall but also an activation of gas exchange and a rise in the content of sugars (sucrose, fructose, and glucose). In sterile clone, degradation of tapetum and anomalies in the development of sporogenic tissue were simultaneously observed in the prophase of meiosis. The death of microsporocytes and degeneration of tapetum were accompanied by a decrease in the level of sucrose delivered to the anther tissues and changes in the ratio between sucrose and hexoses in favor of glucose.