七叶树小孢子发生及雄配子体发育研究

用石蜡切片法观察了七叶树花药的发育过程.结果表明:(1)雄蕊花药四室,花药壁完全分化时,从外到内依次是表皮、药室内壁、中层和绒毡层,花药壁发育为基本型.表皮细胞1层,发育过程中始终存在;药室内壁在花药成熟时形成带状纤维层加厚;幼小花药壁的中层3~4层细胞,在花药发育成熟时退化消失;绒毡层1层细胞,发育类型为分泌型,小孢子母细胞减数分裂时绒毡层开始退化解体,花药成熟完全消失,仅剩1层绒毡层膜.每一花药中有多列雄性孢原细胞,发生于幼小花药表皮下方;(2)小孢子母细胞减数分裂为同时型,四分体多呈正四面体排列;减数分裂过程中,小孢子母细胞外方被胼胝质壁所包被,小孢子形成后胼胝质壁逐渐消失.成熟花粉二细胞型,外形呈圆三角状,具三孔沟.     

Receptor-like protein kinase 2 (RPK 2) is a novel factor controlling anther development in Arabidopsis thaliana

Receptor-like kinases (RLK) comprise a large gene family within the Arabidopsis genome and play important roles in plant growth and development as well as in hormone and stress responses. Here we report that a leucine-rich repeat receptor-like kinase (LRR-RLK), RECEPTOR-LIKE PROTEIN KINASE2 (RPK2), is a key regulator of anther development in Arabidopsis. Two RPK2 T-DNA insertional mutants (rpk2-1 and rpk2-2) displayed enhanced shoot growth and male sterility due to defects in anther dehiscence and pollen maturation. The rpk2 anthers only developed three cell layers surrounding the male gametophyte: the middle layer was not differentiated from inner secondary parietal cells. Pollen mother cells in rpk2 anthers could undergo meiosis, but subsequent differentiation of microspores was inhibited by tapetum hypertrophy, with most resulting pollen grains exhibiting highly aggregated morphologies. The presence of tetrads and microspores in individual anthers was observed during microspore formation, indicating that the developmental homeostasis of rpk2 anther locules was disrupted. Anther locules were finally crushed without stomium breakage, a phenomenon that was possibly caused by inadequate thickening and lignification of the endothecium. Microarray analyses revealed that many genes encoding metabolic enzymes, including those involved in cell wall metabolism and lignin biosynthesis, were downregulated throughout anther development in rpk2 mutants. RPK2 mRNA was abundant in the tapetum of wild-type anthers during microspore maturation. These results suggest that RPK2 controls tapetal cell fate by triggering subsequent tapetum degradation, and that mutating RPK2 impairs normal pollen maturation and anther dehiscence due to disruption of key metabolic pathways.     

Secretory tapetum of Brassica oleracea L.: polarity and ultrastructural features

Summary The ultrastructure of the secretory, binucleate tapetum of Brassica oleracea in the micro spore mother cell (MMC) stage through to the mature pollen stage is reported. The tapetal cells differentiate as highly specialized cells whose development is involved in lipid accumulation in their final stage. They start breaking down just before anther dehiscence. Nuclei with dispersed chromatin, large nucleoli and many ribosomes in the cytoplasm characterize the tapetal cells. The wall-bearing tapetum phase ends at the tetrade stage. The dissolution of tapetal walls begins from the inner tangential wall oriented towards the loculus and proceeds gradually along the radial walls to the outer tangential one. The plasmodesmata transversing the radial walls between tapetal cells persist until the mature microspore, long after loss of the inner tangential wall. After wall dissolution, the tapetal protoplasts retain their integrity and position within the anther locule. The tapetal cell membrane is in direct contact with the exine of the microspores/pollen grains and forms tubular evaginations that increase its surface area and appear to be involved in the translocation of solutes from the tapetal cells to the microspores/ pollen grains. The tapetal cells exhibit a polarity expressed by spatial differentiation in the radial direction.     

Anther, pollen and tapetum development in safflower, Carthamus tinctorius L

In safflower, the anther wall at maturity consists of a single epidermis, an endothecium, a middle layer and the tapetum. The tapetum consists mainly of a single layer of cells. However, this single-layer appearance is punctuated by loci having ‘two-celled’ groupings due to additional periclinal divisions in some tapetal cells. Meiotic division in microsporocytes gives rise to tetrads of microspores. The primexine is formed around the protoplasts of microspores while they are still enveloped within the callose wall. Just prior to microgametogenesis, the microspores enlarge through the process of vacuolation, and the exine wall pattern becomes established. Microgametogenesis results in the formation of 3-celled pollen grains. The two elongated sperm cells appear to be connected. The exine wall is highly sculptured with a distinct tectum, columellae, a foot layer, an endexine and a thin intine. Similar to other members of the Asteraceae family, the tapetum is of the invasive type. The most novel finding of this study is that in addition to the presence of invasive tapetal cells, a small population of ‘non-invasive’ tapetal cells is also present. The tapetal cells next to the anther locules in direct contact with the microspores become invasive and start to grow into the space between developing microspores. These tapetal cells synthesize tryphine and eventually degenerate at the time of gametogenesis releasing their content into the anther locules. A smaller population of non-invasive tapetal cells is formed as a result of periclinal divisions at the time of tapetum differentiation. These cells are not exposed to the anther locules until the degeneration of the invasive tapetal cells. The non-invasive tapetal cells have a different cell fate as they synthesize pollenkitt. This material is responsible for allowing some pollen grains to adhere to each other and to the anther wall after anther dehiscence. This observation explains the out-crossing ability of Carthamus species and varieties in nature.     

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 of thermosensitive genic male-sterile rice is associated with premature programmed cell death of the tapetum

The tapetum plays a crucial role in pollen development. This secretory tissue produces numerous nutritive proteins necessary for pollen maturation. The tapetum, whose cells undergo programmed cell death (PCD), is completely diminished by the time the pollen is fully mature. Our previous studies on a thermosensitive genic male-sterile (TGMS) rice (Oryza sativa L.) suggested that male-sterility was due to failure in pollen development. In this paper we describe how further analysis of the TGMS rice revealed that male-sterility is associated with premature PCD of the tapetum. Cytological observations of TGMS rice anthers at various developmental stages indicated that PCD initiates at an early stage of pollen development and continues until the tapetal cells are completely degraded, resulting in pollen collapse. Transmission electron microscopy showed the morphologically distinct hallmarks of apoptosis, including cytoplasmic shrinkage, membrane blebbing, and vacuolation. Identification of DNA fragmentation using the TUNEL assay supports the hypothesis that premature PCD is associated with male-sterility in the rice. The tissue-specific feature of the thermosensitive genic male-sterile phenotype is discussed with regard to PCD during anther development.     

侧柏小孢子囊壁绒毡层和中层细胞的发育

侧柏[Platycladus orientalis (L.)Franco]小孢子囊壁包括3层细胞：表皮、中层和绒毡层。中层细胞为1层扁平的细胞。绒毡尾细胞属于分泌型。成熟的绒毡层细胞除了有单核和双核细胞外,还有三核和四核等多核细胞,细胞核有圆形和长椭圆形2种形态。绒毡层细胞的洒色质伴随着小孢子母细胞减数分裂有一个浓缩和伸展的时期,这个时期影响营养物质向小孢子囊内部转运,绒毡层细胞发育的初期就为造孢细胞提供营养,后期解体时,分泌的乌氏体不是散乱地而是有组织地向花粉粒的表面转移。中层和绒毡层细胞最终作为营养被全部吸收利用。     

Anther wall layers control pollen sugar nutrition inLilium

Summary Anthers ofLilium were for the first time investigated at the ultrastructural level in order to appreciate the possible ways of sugar transport in the microsporangium. Our results have shown that the cells of the outer anther wall layers and the cell of the connective were interconnected by plasmodesmata, thus allowing assimilates to travel through the symplasmic pathway from the vascular bundle to the most internal middle layer (ML 1). ML 1 was devoid of cell communication throughout pollen development. Tapetal cells were also lacking plasmodesmata on their external face towards ML 1, but adjacent tapetal cells developed lateral junctions: the tapetum could represent a syncytium. Sugars destinated to pollen in the loculus have then to cross the ML 1 and the tapetal layers by the apoplasmic pathway; it is suggested that these two envelopes could be involved in the control of sugar transport from the outer anther wall layers to the locular fluid. Before microspore mitosis, the tapetum degenerated but ML 1 remained structurally unchanged. During pollen development, the guard cells of stomata were lacking cell communication, and preserved their starch content, which could be the sign of photosynthesis within the anther wall. In order to check whether these structural disconnections in anther tissues corresponded to physiological barriers, isolated pollen and stamens were cultivated during the anther maturation phase, on a medium containing increasing concentrations of sucrose (0 M, 1/6 M, 1/2 M, 1 M). After 7 days of culture, isolated pollen was engorged with starch grains and was unable to germinate, whereas in cultivated stamens, pollen did not contain any starch grain: sporophytic tissues, however, accumulated abnormal amylaceous reserves. These results strongly suggest that the anther wall layers, in particular ML 1, starve pollen with sugars during its maturation. They are acting as a physiological buffer storing nutriment surplus in starch grains.Abbreviations ML 1 middle layer 1 - ML 2 middle layer 2 - PAS periodic acid Schiff - PATAg periodic acid thiosemicarbazide silver nitrate     

Anther ontogeny in Campsis radicans (L.) Seem. (Bignoniaceae)

In this study anther ontogeny of Campsis radicans (L.) Seem. was investigated by transmission electron microscopy and light microscopy with special reference to the development of the anther wall. The anther wall formation follows the dicotyledonous type. The differentiation in anther starts with the appearance of archesporial cells which undergo periclinal divisions to give primary parietal layer to the epidermal site and the primary sporogenous cells to the inside. The primary parietal layer also divides to form two secondary parietal layers. Later, the outer secondary parietal layer (spl1) forms the endothecium and the middle layer by periclinal division whereas the inner one (spl2) directly develops into the outer tapetum forming the inner most layer of the anther wall. The sporogenous tissue is generally organized in two rows of cells with a horseshoe-shaped outline. The remainder of the tapetum lining the sporogenous mass is derived from the connective tissue. The tapetum thus has dual origin and dimorphic. Anthers are tetrasporangiate. The wall of the anther consists of an epidermis, endothecium, middle layer, and the secretory type tapetum. Tapetal cells are usually binucleated. Epidermis and Endothecium layers of anther wall remain intact until the end of anther and pollen development; however, middle layer and tapetum disappear during development.     

A contribution to the embryology of Desmodium motorium (Houtt.) Merr. (= D. gyrans (L.f.) DC.) (Fabaceae)

Abstract

The anthers are tetrasporangiate. The anther wall comprises epidermis, fibrous endothecium, middle layer and tapetal layer. The tapetum is of the Glandular type and its cells remain uninucleate. Meiosis in pollen mother cells is normal and simultaneous cytokinesis leads to the formation of tetrahedral and decussate microspore tetrads. The pollen grains are shed at 2-celled stage. The ovule is campylotropous, bitegmic and crassinucellate. Meiosis in megaspore mother cell results in the formation of linear or occasionally T-shaped megaspore tetrad. The chalazal megaspore develops into Monosporic Polygonum type of embryo sac. Endosperm development is of the Nuclear type.     

Pollen development of Cardiocrinum giganteum (Wall.) Makina in China

In this article, we studied the pollen morphology and wall development, microsporogenesis, male gametophyte development, and anther wall structure changes during pollen development of Cardiocrinum giganteum (Wall.) Makina from the genus Cardiocrinum (Endl.) Lindl. (Liliaceae) using paraffin sections, scanning and transmission electron microscopy, and fluorescence microscopy. The results showed that C. giganteum has oval-shaped pollen with a single sulcus and reticulate ornamentation. The exine is of the semi-tectum type and can be divided into the tectum layer, columellate layer and basal layer. Meiosis in the microsporocyte is accompanied by successive cytokinesis. The mature pollen is three-celled. The anther wall prior to maturity is built by one layer of epidermis, 1–2 layers of endothecium cells, 4–5 middle layers and 2 layers of tapetum, while upon maturity it is only built by one layer of epidermis, one layer of endothecium cells and one middle layer. The tapetal cells are secretory, with two or more nuclei. Ubisch bodies originate from rough endoplasmic reticulum except a few from mitochondria.     

MS32-regulated timing of callose degradation during microsporogenesisin Arabidopsis is associated with the accumulation of stacked rough ER in tapetal cells

  In the male sterile32(ms32)mutant in Arabidopsis thaliana, pollen development is affected during meiosis of pollen mother cells (PMCs). In normal wild-type (WT) anthers, callose is deposited around PMCs before and during meiosis, and after meiosis the tetrads have a complete callose wall. In ms32, PMCs showed initial signs of some callose deposition before meiosis, but it was degraded soon after, as was part of the cellulosic wall around the PMCs. The early dissolution of callose in ms32 was associated with the occurrence of extensive stacks of rough ER (RER) in tapetal cells. The stacks of RER were also observed in the WT tapetum, but at a later stage, i.e., after the tetrads were formed and when callose is normally broken down for release of microspores. Based on these observations it is suggested that: (1) callose degradation around developing microspores is linked to the formation of RER in tapetal cells, which presumably synthesize and/or secrete callase into the anther locule, and (2) mutation in MS32 disrupts the timing of these events. Received: 27 April 1999 / Revision accepted: 21 June 1999     

青葙花药发育的结构和组织化学观察

对苋科植物青葙Celosia argentea花药发育的结构和组织化学（多糖和脂滴）特征进行观察。青葙小孢子发生为同时型,四分体为四面体型。药壁为典型四层,绒毡层属于同型绒毡层。成熟花粉为二胞型。早期花药中的淀粉粒和脂滴均较少,绒毡层细胞至小孢子晚期退化为体积较大的脂块。二胞花粉时期的中层细胞退化为脂滴。早期二胞花粉中先出现多糖颗粒,晚期的成熟花粉中积累大量淀粉粒和较少的脂滴为营养储存物。     

凤仙花花药发育中多糖和脂滴组织化学研究

以不同发育时期的凤仙花花药为实验材料,采用组织化学方法,对花药发育中的结构变化及多糖和脂滴物质分布进行观察。结果表明:(1)凤仙花的花药壁由6层细胞组成,包括1层表皮细胞,2层药室内壁细胞,2层中层细胞和1层绒毡层细胞。其中绒毡层细胞的形态不明显,很难与造孢细胞区分,且在小孢子母细胞时期退化。(2)在小孢子母细胞中出现了一些淀粉粒,但减数分裂后,早期小孢子中的淀粉粒消失,又出现了一些小的脂滴;随着花粉的发育,小孢子形成大液泡,晚期小孢子中的脂滴也消失;小孢子分裂形成二胞花粉后,营养细胞中的大液泡降解、消失,二胞花粉中又开始积累淀粉;接近开花时,成熟花粉中充满细胞质,其中包含了较多的淀粉粒和脂滴。(3)在凤仙花的花药发育中,绒毡层细胞很早退化,为小孢子母细胞和四分体小孢子提供了营养物质;其后的中层细胞退化则为后期花粉发育提供了营养物质。     

山麦冬大小孢子发生及雌雄配子体发育研究

采用石蜡切片技术对百合科植物山麦冬大小孢子发生及雌雄配子体发育进行了观察研究。结果表明:(1)山麦冬花药具有4个花粉囊,花药壁的发育方式为基本型,花药壁完全分化时由表皮、药室内壁、中层及绒毡层组成。(2)绒毡层发育类型为分泌型,到四分体孢子彼此分离形成单细胞花粉阶段,绒毡层细胞开始解体退化,花粉成熟时绒毡层细胞完全消失;花粉母细胞减数分裂为连续型,四分体为左右对称形排列,成熟花粉为3-细胞花粉,单萌发沟。(3)子房3室,每室2枚胚珠,胚珠倒生型,双珠被,薄珠心,雌性孢原细胞不经过平周分裂而直接发育而成大孢子母细胞。(4)减数分裂后四分体大孢子呈线型或T型排列,合点端大孢子分化为功能大孢子,胚囊发育为蓼型;花粉母细胞减数分裂过程中,二分体、四分体细胞外方被胼胝质壁所包被,小孢子形成后胼胝质壁逐渐消失。该研究结果丰富了百合科植物生殖生物学研究的内容,也为探讨百合科植物的系统学研究提供了参考。     

A comparative light and electron microscopic analysis of microspore and tapetum development in fertile and cytoplasmic male sterile radish

To gain further insight into the abortive stages and ultrastructural changes leading to pollen degeneration of a novel cytoplasmic male sterile radish 805A, we compared differences of cellular and subcellular structure of sterile anther with fertile anther by light and electron microscopy analysis. Two types of locule degeneration in sterile anther were detected, of which the time of degeneration occurred and completed was different. In type I, abnormality of pollen mother cells (PMCs) and tapetal cells, including condensation of cytoplasm and large vacuoles within tapetal cells, was shown at PMC stage. In type II, meiosis and early tetrad stage progressed normally except for large vacuoles that appeared in tapetal cells. Ultrastructural alterations of the cellular organization were observed in the type II locules, such as chromatin condensation at the periphery of the nucleus and degeneration of the karyotheca, compared with normal pollen development. The results suggested that the cytoplasmic male sterility anther degeneration was probably caused by dysfunctions of tapetum and vacuolation of tapetum, PMCs, and microspores. Thus, the identical factors, which induced CMS in the same cytoplasmic and nuclear genetic background, might affect development of tapetum and microspore at different stages during the cytoplasmic male sterile 805A anther development.     

百合花药壁层的发育及组织化学研究

对生长在陕西留坝的百合的花药壁层发育过程,特别是绒毡层的发育做了形态学观察。其结果是：百合花药壁层的发育方式为基本型。花药绒毡层属腺质绒毡层类型。在单细胞花粉阶段后期,部分花粉粒壁一侧凹陷时,绒毡层细胞内切向面上出现乌氏体。随着发育阶段的推移,乌氏体的数量有所增加。在光学显微镜下观察：每个乌氏体只有一个乌氏体芯。在乌氏体出现时,也可观察到花粉外壁外层的出现。到二细胞花粉时,花药开裂之前,绒毡层细胞     

PERSISTENT TAPETAL CELL1 encodes a PHD-finger protein that is required for tapetal cell death and pollen development in rice

In higher plants, timely degradation of tapetal cells, the innermost sporophytic cells of the anther wall layer, is a prerequisite for the development of viable pollen grains. However, relatively little is known about the mechanism underlying programmed tapetal cell development and degradation. Here, we report a key regulator in monocot rice (Oryza sativa), PERSISTANT TAPETAL CELL1 (PTC1), which controls programmed tapetal development and functional pollen formation. The evolutionary significance of PTC1 was revealed by partial genetic complementation of the homologous mutation MALE STERILITY1 (MS1) in the dicot Arabidopsis (Arabidopsis thaliana). PTC1 encodes a PHD-finger (for plant homeodomain) protein, which is expressed specifically in tapetal cells and microspores during anther development in stages 8 and 9, when the wild-type tapetal cells initiate a typical apoptosis-like cell death. Even though ptc1 mutants show phenotypic similarity to ms1 in a lack of tapetal DNA fragmentation, delayed tapetal degeneration, as well as abnormal pollen wall formation and aborted microspore development, the ptc1 mutant displays a previously unreported phenotype of uncontrolled tapetal proliferation and subsequent commencement of necrosis-like tapetal death. Microarray analysis indicated that 2,417 tapetum- and microspore-expressed genes, which are principally associated with tapetal development, degeneration, and pollen wall formation, had changed expression in ptc1 anthers. Moreover, the regulatory role of PTC1 in anther development was revealed by comparison with MS1 and other rice anther developmental regulators. These findings suggest a diversified and conserved switch of PTC1/MS1 in regulating programmed male reproductive development in both dicots and monocots, which provides new insights in plant anther development.     

水稻胞质雄性不育系珍汕97A及其保持系珍汕97B绒毡层发育过程中的Ca2+分布变化

利用焦锑酸钾沉淀法研究了野败不育系珍汕97A及其保持系珍汕97B绒毡层细胞的发育过程及其细胞中Ca2 的分布变化。研究发现保持系绒毡层细胞在单核花粉晚期才开始迅速解体,而不育系绒毡层细胞在花粉母细胞时期就开始出现核膜、细胞膜解体,此过程持续到二核花粉时期。珍汕97A绒毡层细胞从花粉母细胞时期开始,细胞质内有少量颗粒状的Ca2 沉淀;减数分裂时期,绒毡层细胞的内切向壁表面有大量大颗粒的Ca2 沉淀;单核花粉时期绒毡层细胞周围集聚一层Ca2 沉淀。而保持系绒毡层细胞遮花粉母细胞时期和减数分裂时期细胞内没有Ca2 沉淀;单核花粉时期绒毡层细胞内的Ca2 沉淀主要分布在解体的细胞质内。推测绒毡层细胞结构发育的异常和Ca2 的异常分布可能与花粉的败育有关。     

铁皮石斛花粉块结构及发育过程的解剖学特征研究北大核心CSCD

兰科植物的有性生殖特殊,每朵花只有1个花药,且花粉有聚集成块发育的特征。为了揭示铁皮石斛花粉块的发育特征,该研究以野生铁皮石斛不同时期的花药为材料,采用半薄切片和植物组织化学方法对其发育过程进行解剖学观察分析,并对成熟花粉块进行离体培养,观察花粉管的萌发状况。结果表明:(1)铁皮石斛花药壁由1层表皮细胞,2层药室内壁细胞,1层中层细胞和1层绒毡层细胞组成。开花时,绒毡层细胞退化,中层细胞没有退化,药室内壁细胞则形成纤维状细胞壁;药室中的小孢子母细胞没有明显的胼胝质壁结构。(2)小孢子发生属同时型,减数分裂后四分体小孢子不分散,以四合花粉状态发育,并进一步连接形成花粉块。(3)在小孢子发育中,孢粉素覆盖在整个花粉块表面形成花粉外壁,但花粉块内部的花粉没有花粉外壁结构;在花粉块表面的花粉外壁上未见花粉萌发孔。(4)在花粉离体萌发实验中,具有花粉外壁的花粉块表面花粉未见萌发,仅由花粉块内部的花粉萌发出花粉管。