DNA synthesis and cytoplasmic differentiation in tapetal cells of normal and cytoplasmically male sterile lines of Petunia hybrida

Summary A new method is described by which tapetal cells may be isolated from anthers of cytoplasmic male sterile (CMS) and fertile lines of Petunia hybrida. Using a combination of stereometry and Feulgen densitometry it has been possible to chart DNA synthesis and nuclear division with great precision within these cells. Results are presented which show CMS lines both to synthesize far less DNA than the fertiles and also to undergo less cell division. These differences in DNA kinetics and cytokinesis are obvious long before any differences between sterile and fertile lines may be detected in the meiocyte mass. In addition to these difference in nuclear behaviour, the tapetal cells of CMS lines also synthesize far lower levels of rough endoplasmic reticulum than do their fertile counterparts. Again, this difference is evident at a very early stage of anther development when all other cell components, including mitochondria and plastids, appear normal. These data are discussed in terms of the very special differentiation undergone by tapetal cells in angiosperms, and the conclusions drawn are considered in the perspective of current hypotheses proposed to explain the operation of CMS.     

白头翁绒毡层和周绒毡层膜的发育和组织化学研究

白头翁绒毡层为分泌型,发育过程分为：发生－分化,生长－合成和分泌－解体3个阶段。第1阶段,绒毡层细胞的形态结构和组织化学特征同造孢组织和花粉母细胞的大体一致,所不同的是大量积累淀粉和形成少量原乌氏体,第2阶段为重要的合成时期;细胞体积膨大,具多倍体双核或畸形核,蛋白质大量合成,淀粉水解;原乌氏体和孢粉素荧光物质大量累积。第3阶段为分泌最活跃时期,裸细胞在妨行分泌功能上起着重要作用。原乌氏体,荧光物质,碳水化合物,或许包括分解胼胝质壁和纤维素壁酶等成分均在裸细胞阶段排出,周绒毡层膜,乌氏体和花粉外壁均属于孢粉素性质的结构,根据周绒毡层膜的外切向位置和结构,初步结论：它可能是绒毡层细胞质膜向着药壁中层一面的残存部分,而膜内表面上的孢粉素纹理则是后来添加的。乌氏体的基本形态为短颈烧瓶状,单个或2－5个组成复合结构,本文对绒毡层,绒毡层膜和乌氏体的功能,及孢粉素的转运和聚合等问题进行了讨论。     

杉木雄性不育株与可育株小孢子囊发育的电镜研究

杉木雄性不育属“无花粉型”,败育从无孢原细胞到四分体时期,中层细胞增生,压迫小孢子母细胞,使之养分更加缺乏并引起减数分裂异常。其表皮层和药室内壁细胞中具大量蛋白体,影响了绒毡层对小孢子发育的外源蛋白的供应;表皮层具膜状溶酶体,引起淀粉粒缺乏和酶系统代谢异常;绒毡层发育后期,缺少包被小泡、乌氏体及绒毡层膜,内质同少、短而光滑,严重影响了绒毡层对小孢子母细胞或小孢子养分和合成细胞壁物质的供给;线粒体发育异常,能量代谢减弱,导致小孢子母细胞或小孢子一系列生理活动紊乱及其原生质体解体。     

杉木雄性不育株与可育株小孢子囊发育的电镜研究

杉木雄性不育属“无花粉型”,败育从无也原细胞押分体时期、中层细胞增生,压迫小孢子母细胞,使之养分更加缺乏并引起减烽分裂异常。其表皮层和药室内壁细胞中具大量蛋白体,影响了绒毡层对小孢子发育的外源蛋白的供应;表皮层具膜状溶酶体,引起淀粉粒缺乏和酶系统代谢异常;绒毡层发育后期,缺少包被小泡,乌氏体及绒毡层膜,内质网少,短而光滑,严重影响了绒毡层对小孢子母细胞或小孢子养分和合成细胞壁物质的供给;     

The loculus content and tapetum during pollen development in Lilium

 The ratio of loculus volume to the volume of the entire anther began to increase from the microspore mother cell stage and reached 32.3% at anthesis. The content of the loculus was examined in Lilium during pollen development and two waves could be distinguished. From the premeiotic stage until the vacuolated microspore stage, the loculus consisted of neutral polysaccharides, pectins and proteins. These substances originated from tapetal activity from the premeiotic stage until the young microspore stage. Dictyosomes and rough endoplasmic reticulum seemed to be involved in tapetal secretion, although, in some mitochondria, vesicles progressively developed as early as premeiosis and increased until the young microspore stage, which could reveal their involvement in the secretion process. At this stage, numerous cytoplasmic vesticles containing material similar to the locular material fused with the plasma membrane of the tapetum so that vesicle content was in contact with the loculus. It seems that tapetal and callose wall degradation at the late tetrad stage may also have contributed to the production of material in the loculus. From pollen mitosis to anthesis, the anther loculus contained mainly the pollenkitt which was synthesized in the tapetum between the young microspore stage and the vacuolated microspore stage. At the young microspore stage, proplastids divided and developed into elaioplasts and smooth endoplasmic reticulum (SER) increased dramatically. Pollenkitt had a double origin: some droplets were extruded directly from the plastid stroma through the plastid envelopes; the others were unsaturated lipid globules, which presumably derived from the interaction between SER saccules and plastids. Received: 2 September 1997 / Revision accepted: 12 March 1998     

Pollen ontogenesis in Oenothera: a comparison of genotypically normal anthers with the male-sterile mutant sterilis

Summary A 12-stage normal table of anther development in Oenothera, is presented. The stages are characterized by developmental steps in the reproductive cells and the tapetum, including waves of amylogenesis and lipogenesis as well as the production of the sporoderm layers. This is compared to a corresponding table for the male-sterile (mst) mutant sterilis (ster). Differences between the development of fertile and mst anthers appear after the liberation of the microspores from the tetrads. Male sterility results from a malfunction of the tapetum in the production of ektexine sporopollenin precursors, which aggregate in the tapetal cells. The consequence is the absence of ektexine from the microspores. The endexine is then dissolved, presumably by an enzyme. This process leads to naked microspores whose unprotected cytoplasms are attacked by hydrolytic enzymes present in the thecal fluid. At anthesis the anthers contain only undefined remnants of microspores and tapetum.     

ST273 Is Essential for Tapetum and Microspore Development in Arabidopsis thaliana*

In Arabidopsis, the tapetum plays important roles in anther and pollen development by providing enzymes for callose dissolution, materials for pollen wall formation, and nutrients for microspore development. This paper describes the functional analyses of the ST273 gene in anther and pollen development by using Arabidopsis male sterile mutant st273. Mutant st273 was identified from a T DNA insertion mutant population, and genetic analysis showed that st273 mutant was controlled by a single recessive nuclear gene. A map based cloning approach was used, and ST273 gene was mapped to be linked to a molecular marker CIW11 on chromosome 3. Bioinformatics analysis revealed that there is a TDF1 gene near the marker CIW11. Sequencing analysis indicated that st273 mutant had a G459 to A459 base pair change in the third exon of TDF1 gene, which resulted in premature termination mutation in this region. Allelism test indicated that ST273 and TDF1 belong to the same locus. The mutant plant grows normally during the vegetative growth stage, but show developmental defects at the reproductive growth stage. Alexander staining showed that there was no pollen in the mature anther locule. Cytology observation indicated that the mutant tapetum was enlarged and vacuolated, the tetrads could not release the microspores timely, and finally no pollen was formed in the anther. These results demonstrated that ST273 protein plays an important role in tapetum and microspore development.     

Ultrastructural study on degeneration of tapetum in anther of snap bean (Phaseolus vulgaris L.) under heat stress

Pollen sterility was induced by heat stress applied about 10 days before flowering in the snap bean Phaseolus vulgaris L. Cytohistological changes in the tapetum during early development of the anther were studied to identify the tissues most sensitive to high temperature stress. The first distinct structural abnormalities were detected in the distribution pattern of the rough endoplasmic reticulum (RER) in the tapetum at the early microspore stage under high temperature conditions. Stacks of RER were frequently observed in the tapetum under optimal conditions, but rarely occurred under high temperature conditions. Various patterns of endoplasmic reticulum (ER) arrangement – linear, wavy, looped or circular – were observed in the tapetum. Two types of circular ER were observed at the microspore stage under high temperature conditions, RER with ribosomes on the surface and smooth endoplasmic reticulum (SER) lacking ribosomes. The tapetum underwent degenerative changes under high temperature conditions earlier than under optimal conditions. The structural abnormalities of the microspore were associated with tapetal degeneration. We concluded that high air temperature affected the ER structure and blocked its function in the tapetum, and then induced earlier than usual degeneration of tapetum. Pollen sterility is associated with tapetal degeneration. Received: 12 October 2000 / Revision accepted: 7 March 2001