Calcium distribution in fertile and sterile anthers of a photoperiod-sensitive genic male-sterile rice

Potassium antimonate was used to locate Ca²⁺ in fertile and sterile anthers of a photoperiod-sensitive genic male-sterile rice (Oryza sativa L. japonica). During the development of fertile anthers, abundant calcium precipitates accumulated in the anther walls and on the surface of pollen grains and Ubish bodies at the late developmental stage of the microspore, but not in the cytoplasm of pollen grains. Following the accumulation of starch grains in pollen, calcium precipitates on pollen walls diminished and increased in parenchymatous cells of the connective tissue. In sterile anthers, calcium precipitates were abundant in the middle layer and endothecium, but not in the tapetum, as was found in fertile anthers. A special cell wall was observed between the tapetum and middle layer of sterile anthers that appeared to relate to distinctive calcium accumulation patterns and poor pollen wall formation in the loculi. The formation of different patterns of antimonate-induced calcium precipitates in the anthers of photoperiod-sensitive genic male-sterile rice indicates that anomalies in the distribution of calcium accumulation correlate with the failure of pollen development and pollen abortion. Received: 30 May 1997 / Accepted: 5 July 1997     

云南紫稻细胞质雄性不育系花药发育过程中Ca2+的分布特点

运用焦锑酸钾沉淀法研究了云南紫稻细胞质雄性不育系和保持系花药在发育过程中Ca^2 的分布特点。结果表明,保持系的花粉母细胞和小孢子的胞质内部基本无Ca^2 的沉淀,后期花粉外壁出现Ca^2 的沉淀;保持系早期的绒毡层细胞形态正常,胞内有少量Ca^2 沉淀,后期绒毡层细胞开始凋亡,胞质凝集,胞内出现大量Ca^2 的颗粒。不育系花粉母细胞在减数分裂时期败育,胞质液泡化,内部出现大量Ca^2 的沉淀;不育系绒毡层细胞形态正常,胞内无Ca^2 的沉淀。绒毡层与花粉母细胞、小孢子之间出现大量Ca^2 颗粒。探讨了不育系花药花粉母细胞中以及与绒毡层细胞之间Ca^2 的异常积累与雄性不育的关系。     

水稻非花粉型细胞质雄性不育系及其保持系花药发育过程中Ca2+的分布变化

钙在高等植物中被称为第二信使,与植物的有性生殖有关。为了研究水稻（Oryza sativa L.）花药中钙的定位与花粉败育的关系,利用焦锑酸钾沉淀法研究了非花粉型细胞质雄性不育系G37A及其保持系G37B花药的发育过程及其细胞中Ca^2＋ 的分布变化。研究发现,在2个材料间花药中钙的分布存在大量差异。G37B的可育花药在花粉母细胞时期及二分体时期,很少看到有Ca^2＋的沉积;而在单核花粉时期,Ca^2＋沉积急速地增加,主要定位在绒毡层细胞、花粉外壁外层及乌氏体的表面;随后花药壁上沉积的Ca^2＋减少而花粉的外壁外层仍然有很多Ca^2＋沉积物。相反,G37A的不育花药在花粉母细胞时期和二分体时期有大量的Ca^2＋沉积在小孢子母细胞和花药壁,中间层和绒毡层特别多。在二分体时期之后,不育花药的Ca^2＋沉积减少,特别是绒毡层内切向质膜附近的Ca^2＋几乎消失。但是同时期的可育花药中,有大量的Ca^2＋沉积在绒毡层。不育花药的Ca^2＋沉积在开花几天后消失。根据研究结果推测在不育花药发育早期中更多的钙离子与花粉败育有一定的关系。     

温敏雄性不育水稻培矮64S花药发育过程中钙的变化

采用焦锑酸钾沉淀法研究了温敏雄性不育水稻(Oryza sativa L.)培矮64S在高温引起雄性不育与正常可育花药发育过种中Ca2+的分布变化.结果表明,当培矮64S生长在较高温度条件下引起雄性不育,与可育花药相比,不育花粉母细胞中有较多的液泡、较多的Ca2+沉积和较少的线粒体,并且有较多的Ca2+沉积在不育花药的中间层、表皮层和绒毡层中.到四分体与单细胞花粉时期,不育花药的木质部细胞的次生加厚壁上有较多的Ca2+沉淀,连接组织中的Ca2+沉淀也大大增加,所有不育花粉外壁较厚而发育都不正常.在单核细胞早期,不育花粉的四分体细胞中有较明显的大液泡出现.不育花药中的Ca2+在花药发育的各时期均比可育花药要多.这些结果说明在高温生长条件下,花粉母细胞发育的异常、花药中Ca2+沉积的增加、绒毡层与花粉外壁发育的异常可能与培矮64S花粉败育相关.     

白菜细胞核雄性不育花药的细胞化学观察

对一种由一对隐性基因控制的白菜细胞核雄性不育和可育株的花药进行了细胞学和组织化学研究。种子播种后,有1／4植株为不育株,其余的为可育株。通过对不育株和可育株花药发育的细胞学观察,确认不育花粉的败育发生在小孢子发育时期。用组织化学的方法研究了可育株和不育株花药发育过程中的多糖和脂类的分布动态,发现在减数分裂前,可育花药和不育花药的药隔细胞中都储藏了大量的淀粉粒。二者的差异仅是不育花药的绒毡层细胞液泡化明显。在减数分裂后的小孢子发育时期,可育花药的绒毡层细胞具有将药隔细胞中的淀粉粒多糖吸收并转化成脂类的功能,小孢子及以后的二胞花粉中也积累了大量的脂类储藏物质。在不育花药中,虽然减数分裂后药隔细胞中的淀粉粒也都消失,但绒毡层细胞中的脂类物质相比很少,同时绒毡层细胞显示了明显的多糖反应,表明不育花药的绒毡层细胞将糖类转化为脂类的功能受阻。在小孢子的表面有些脂类物质,但在细胞质中却没有脂类积累。这一结果暗示在该种白菜细胞核雄性不育株中,由于花药绒毡层细胞转换多糖为脂类的功能失常,导致了小孢子的败育。     

白菜核雄性不育花药超微结构的研究

对白菜核雄性不育两用系的可育与不育花药进行了超微结构的比较观察。结果显示不育花药的造孢细胞核仁靠边分布;包裹小孢子母细胞的胼胝质厚薄不均匀,不完整等早期异常现象。减数分裂后．四分体细胞中常有多个细胞核。从四分体释放出的小孢子外壁的孢粉素物质不均匀沉积,呈不连续的单层异常结构。最后小孢子通过细胞质收缩方式败育。在可育花药中．绒毡层细胞在小孢子发育后期已显示出退化迹象,同时在细胞中开始积累脂类物质。但在同时期的不育花药中．绒毡层细胞没有显示出退化的迹象,也不合成脂类物质。从时间上看,败育花药中小孢子母细胞及小孢子的异常在先,绒毡层细胞的异常在后。本研究揭示了白菜核雄性不育花药的超微结构特征．对我们以前的光学显微镜观察结果予以补充和修正。     

白菜核雄性不育花药超微结构的研究

对白菜核雄性不育两用系的可育与不育花药进行了超微结构的比较观察。结果显示不育花药的造孢细胞核仁靠边分布:包裹小孢子母细胞的胼胝质厚薄不均匀,不完整等早期异常现象。减数分裂后,四分体细胞中常有多个细胞核。从四分体释放出的小孢子外壁的孢粉素物质不均匀沉积．呈不连续的单层异常结构。最后小孢子通过细胞质收缩方式败育。在可育花药中,绒毡层细胞在小孢子发育后期已显示出退化迹象,同时在细胞中开始积累脂类物质。但在同时期的不育花药中, 绒毡层细胞没有显示出退化的迹象,也不合成脂类物质。从时间上看,败育花药中小孢子母细胞及小孢子的异常在先,绒毡层细胞的异常在后。本研究揭示了白菜核雄性不育花药的超微结构特征, 对我们以前的光学显微镜观察结果予以补充和修正。     

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。     

Calcium distribution in fertile and sterile anthers of thermosensitive male-sterile wheat

Calcium distribution in fertile and sterile anthers of a thermosensitive male-sterile wheat genotype was investigated using an antimonate precipitation method. During fertile anther development, before meiosis of the microspore mother cells, calcium precipitates were apparent in tapetal cells of the anther wall. After meiosis, precipitates were detected in the early microspores and accumulated in the large vacuole of late microspores. After microspore division, following decomposition of the large vacuole, precipitates decreased in the bicellular pollen. The earliest abnormality in calcium precipitate distribution detected during sterile pollen development was the greater accumulation of precipitates in the cytoplasm and nucleus of late microspores. The sterile microspore can divide to form bicellular pollen, but the large vacuole of sterile bicellular pollen did not decompose and greater abundance of precipitates was retained in the large vacuole. Abnormal distribution of calcium precipitates in sterile pollen precedes structural changes, suggesting that abnormal calcium metabolism is associated with pollen abortion.     

水稻马协不育与可育花药ATPase定位（简报）

Pb3 (po4)2-precipitation was used to study the ATPase activities both in fertile and sterile anthers of rice (Oryza sativa L. cv. Marxie). At big-vacuole pollen stage, tapetum of the fertile anthers showed high ATPase activity in their nuclei. In fertile pollen, ATPase was localized on the outside surface of the exine and in the nucleus both at big-vacuole and at bi-nucleate pollen stage. At late bi-nucleate pollen stage, a large amount of Pb3 (PO4)2 precipitated in endintine of the fertile pollen. In sterile anthers, tapetum was fully degenerated at big-vacuole pollen stage. In sterile pollen, ATPase was localized both on the plasmamembrane and in intine. These phenomena lasted to the bi-nucleate pollen stage. In addition, most of the sterile pollen did not show well-developed endintine. Based on the above results, we suggested that abortive tapetum could not provide enough nutrition for pollen development, and the high ATPase activity both on plasma membrane and in intine would likely result in ATP shortage in sterile anthers.     

白菜核雄性不育系可育和不育花药中Ca2+的分布

研究了白菜(Brassica campestris L. ssp.chinensis Makino)细胞核雄性不育系花药中Ca2 的分布特征.在可育花药发育过程中,减数分裂后花药壁细胞中钙颗粒明显增加.早期小孢子开始积累钙颗粒并特异性地附在小液泡膜上.小孢子分裂后,大液泡消失过程中又伴随着许多钙颗粒附在小液泡膜上,显示出Ca2 与花粉中液泡的形成和分解有关.在不育花药中,最早出现的钙颗粒异常分布是在小孢子母细胞的胼胝质壁中积累了较多的钙颗粒.然而,在小孢子细胞质中钙颗粒一直很少,也不形成大液泡,最后通过细胞质收缩的方式败育.这是首次发现Ca2 参与调控花药发育过程,其异常分布与花粉败育密切相关.     

水稻马协不育与可育花药ATPase定位(简报)

马协不育系系朱英国等用农家品种“马尾粘”的不育株与“协青早选”测交和回交选育成功的。超微结构研究表明马协不育花药单核边位期绒毡层提前解体,液泡膜破裂;不育花粉外壁基粒棒少,且分布不均匀;线粒体有大的电子透明区,二核期不育花粉中央大液泡崩溃。不育花药绒毡层提前解体可能影响花粉外壁合成及花粉营养供应,大量研究表明绒毡层发育异常与花粉败育密切相关,而液泡膜的破坏会导致水解酶类的泄     

Differences on the microsporogenesis and tapetal development of male fertile and cytoplasmic male sterile pepper (Capsicum annuum L.)

To clarify the time and cause of pollen abortion, differences on the microsporogenesis and tapetum development in the anthers of male fertile maintainer line and cytoplasmic male sterile (CMS) line pepper were studied using transmission electron microscopy. The results showed that CMS line anthers appeared to have much greater variability in developmental pattern than male fertile maintainer line ones. The earliest deviation from normal anther development occurred in CMS line anthers at prophase I was cytomixis in some microspore mother cells (MMCs), and vacuolisation in tapetal cells. Then, MMCs in CMS line anthers developed asynchronously and a small part of ones at the different stage degenerated in advance appearing to have typical morphological features of programmed cell death (PCD). Most MMCs could complete the meiosis, but formed non-tetrahedral tetrad microspores with irregular shape and different size and uncertain number of nuclei, and some degenerated ahead of time as well. Tapetal cells in CMS line anther degenerated during meiosis, and were crushed at the tetrad stage, which paralleled the collapse of pollens. Pollen abortion in CMS line anthers happened by PCD themselves, and the premature PCD of tapetal cells were closely associated with male sterility.     

白菜细胞核雄性不育花药的细胞化学观察

对一种由一对隐性基因控制的白菜细胞核雄性不育和可育株的花药进行了细胞学和组织化学研究。种子播种后,有1/4植株为不育株,其余的为可育株。通过对不育株和可育株花药发育的细胞学观察,确认不育花粉的败育发生在小孢子发育时期。用组织化学的方法研究了可育株和不育株花药发育过程中的多糖和脂类的分布动态,发现在减数分裂前,可育花药和不育花药的药隔细胞中都储藏了大量的淀粉粒。二者的差异仅是不育花药的绒毡层细胞液泡化明显。在减数分裂后的小孢子发育时期,可育花药的绒毡层细胞具有将药隔细胞中的淀粉粒多糖吸收并转化成脂类的功能,小孢子及以后的二胞花粉中也积累了大量的脂类储藏物质在不育花药中,虽然减数分裂后药隔细胞中的淀粉粒也都消失,但绒毡层细胞中的脂类物质相比很少,同时绒毡层细胞显示了明显的多糖反应,表明不育花药的绒毡层细胞将糖类转化为脂类的功能受阻。在小孢子的表面有些脂类物质,但在细胞质中却没有脂类积累。这一结果暗示在该种白菜细胞核雄性不育株中,由于花药绒毡层细胞转换多糖为脂类的功能失常,导致了小孢子的败育。     

Involvement of phenylalanine ammonia-lyase in the development of pollen in broccoli (Brassica oleracea L.)

Summary To determine whether phenylalanine ammonia-lyase (EC 4.3.1.5) is involved in the maturation of microspores to fertile pollen, anthers of a fertile strain of broccoli (Brassica oleracea L.) were studied in a comparison with anthers of a cytoplasmic male sterile strain. In the normal fertile strain, immature anthers of about 2 mm in length exhibited higher phenylalanine ammonia-lyase activity than mature anthers or those shorter than 2 mm. The 2-mm-long anthers corresponded to the mononucleate stage, just after release of the microspores during pollen development. Immunohistochemical localization of phenylalanine ammonia-lyase in the anthers indicated that the protein was present predominantly in the tapetal cells. The immature anthers of cytoplasmic male sterile broccoli had a lower phenylalanine ammonia-lyase activity than those of the normal fertile strain. The level of phenylalanine ammonia-lyase activity in the immature anthers was positively correlated with the number of fertile pollen grains at the flowering stage in both strains. It seems possible, therefore, that phenylpropanoid metabolism, which involves phenylalanine ammonia-lyase, may play an important role in the maturation of microspores in flowering plants.Abbreviations CHS chalcone synthase - CMS cytoplasmic male sterility - DAPI 4, 6-diamidmo-2-phenylindole dihydrochloride - PAL L-phenylalanine ammonia-lyase     

新型光温敏小麦不育系337S的组织结构研究

普通小麦(Triticum aestivum)不育系337S是一种对短日低温、长日高温均敏感不育的新型光温敏雄性不育系。对经过短日低温、长日高温处理的不育系花药及其小孢子的形态和发育过程进行了观察,观察结果表明,不育系337S的花药异常短小,开花后花丝短,花药难外露。花药发育过程中中层组织发育紊乱,绒毡层提前解体,影响了花粉母细胞发育所需的营养供应,导致短日低温处理下的花粉母细胞减数分裂中期Ⅰ和长日高温处理下的花粉母细胞发育时期花粉母细胞发育异常,形成异常小孢子,造成败育。     

Cytochemical investigation of genic male-sterility in Chinese cabbage

A genic male sterile Chinese cabbage, Brassica campestris L. ssp. chinensis Makino, was examined using cytological and cytochemical methods to characterize the process of pollen abortion in this plant. Thick sections of both fertile and sterile anthers at different developmental stages were stained using Toluidine Blue O, Periodic Acid-Schiff’s (PAS) reaction and Sudan Black B to detect cytochemical changes that may occur in the distribution of insoluble polysaccharide and lipid storage bodies. Pollen abortion in sterile anthers occurs at an early stage of microspore development. During early microspore development, reductions in the number of starch grains in the connective tissue of fertile anthers coincide with the accumulation of starch grains in cells of the anther wall. In the late microspore stage, a large vacuole forms in the microspore, and tapetal cells synthesize and accumulate lipid droplets. The cellular organization of tapetal cells in sterile anthers appears similar to that in fertile anthers, except for the absence of lipid droplets in cells of sterile anthers and diffusely labeled tapetal polysaccharides, suggesting defects in nutrient storage. Supported by National Natural Science Foundation of CHINA (30170060)     

Developmental and cytogenetic analyses of pollen sterility in Valeriana scandens L.

Valeriana scandens presents perfect and pistillate flowers, the latter with sterile anthers. The species is composed of two varieties with different ploidy; V. scandens var. scandens (2n = 28) and V. scandens var. candolleana (2n = 56), both of which occur in RS, Brazil. Crosses between these varieties may give rise to hybrids with pollen sterility. In this study, we analyzed the microsporogenesis and microgametogenesis of sterile and fertile anthers, and also investigate whether pollen sterility is caused by an irregular meiotic process. Developmental analysis using light microscopy and scanning electron microscopy showed that sterile anthers develop similarly to fertile anthers until the end of meiosis. After this stage, sterile tetrads do not separate as a consequence of exine fusion between adjacent microspores, which is similar to sterile pollen of Brassica ms-cdl1 mutants. In addition, vacuolated immature pollen grains degenerate after separation. The cytogenetic analysis of the microspore mother cell (MMC) showed that the diploid population of V. scandens var. scandens (2n = 28) has pollen sterility that is not caused by a cytogenetic disturbance. The MMCs analyzed from prophase I to tetrad stage showed a regular meiotic process, indicating the phenotype of V. scandens sterile pollen is a postmeiotic process formed by fusion of exine between opposite microspores.     

光（温）敏核雄性不育水稻育性转换中幼穗和花药的某些生理指标变化

光（温）敏核雄性不育水稻（Ｐｈｏｔｏｐｅｒｉｏｄ／ｔｅｍｐｅｒａｔｕｒｅ－ｓｅｎｓｉｔｉｖｅｇｅｎｉｃｍａｌｅ－ｓｔｅｒｉｌｅｒｉｃｅ,简称Ｐ／ＴＧＭＲ）８９０２５和培矮６４ｓ在秋季从不育阶段经半不育阶段向可育阶段转变进程中,对其花粉母细胞形成期（Ⅴ）、花粉母细胞减数分裂期（Ⅵ）的幼穗和成熟花药（Ⅷ）中过氧化物酶（Ｐｅｒｏｘｉｄｓｅ,简称ＰＯＤ）、超氧物歧化酶（Ｓｕｐｅｒｏｘｉｄｅｄｉｓｍｕｔａｓｅ,简称ＳＯＤ）．和过氧化氢酶（Ｃａｔａｌａｓｅ,简称ＣＡＴ）活性测定表明,幼穗和花药相同发育时期的酶活性随正常花粉的比率升高而增加,但以成熟花药中增加最多．并伴有丙二醛（Ｍａｌｏｎａｌｄｅｈｙｄｅ,简称ＭＤＡ）含量减少．表明自由基代谢的变化可能与光（温）敏核雄性不育水稻花粉败育有关,且自由基代谢变化在幼穗发育中期即已开始．     

Effect of Photoperiod on Calcium Distribution in Photoperiod-sensitive Cytoplasmic Male-sterile Wheat During Anther Development

Potassium antimontate was used to locate Ca2+ in fertile and sterile anthers of photoperiod-sensitive cytoplasmic male-sterile wheat (Triticum aestivum L.).The quantity and distribution of calcium precipitates change in microspore and pollen during microsporogenesis and pollen development.The calcium precipitates gradually accumulated on the surface of tetraspore or pollen until pollen mature. They were abundant on the surface of the trinucleate pollen,but not in the cytoplasm.Calcium was transported in anther wall and intervening connective tissue via symplast and apoplast system,meeting the need of microspore and pollen development.Abortive pollens were observed in the anthers developed under long daylight exposure. Abundant calcium precipitates were distributed along the edge of the degradatived pollen plasma and in the locular matrix,but little in the anther wall and intervening connective tissue of the early abortive pollen.Calcium precipitates accumulated on the surface of pollen was less than fertile pollen,but abundant calcium appear in the epiderms,endotheciums,middle layers of anther wall and cells of connective tissue.The results indicate that abnormalities in the distribution of calcium correlate with the failure of pollen development in photoperiod-sensitive cytoplasmic male-sterile wheat.