Cytological Observations on Microsporogenesis and Pollen Development of Regenerated Stamen in Hyacinth

The present study consists of the cytological observations on the process of microsporogenesis and pollen development in the regenerated stamen of hyacinth; and a comparative study of the cytological changes in stamens of both regenerated and produced under natural condition. Results showed that the cytological changes of microsporogenesis and the pollen deveLopment in the regenerated stamen of the hyacinth were basically normal. But in the stage of the mature pollen there was an obvious cytological difference between both stamens in vitro and in nature. The mature pollen of the regenerated stamen consisted of three cells: one vegetative cell and two sperms, while mature pollen grain under natural condition was made up of two cells: one vegetative cell and one generative cell. This difference mainly resulted from different time and place of the generative cell division. The reason resulting in the differences and their influence on sperms were discussed.     

Effects of Temperature on Induction of Regenerative Stamens in vitro Culture,Microsporogenesis and Pollen Development in Hyacinthus orientalis L.

In order to culture the regenerated stamens of hyacinth in vitro to maturity, the effects of temperature on regeneration of stamens, microsporogenesis and pollen development were studied. Results showed: the proper temperature for stamen regeneration was 25℃. The temperature going down gradually was advantageous to the microsporogenesis and pollen development. The most suitable temperatures for differentiation of microsporocyte, meiotic division and pollen first mitosis were 20–25℃, 20℃ and 10℃ respectively. Under such temperature condition, it was possible to culture the regenerated stamens to maturity, and pollen grains in the stamens had the higher germination frequency. On the contrary, unsuitable temperature condition will make microsporogenesis and pollen development stop at certain development stage, it will finally result in pollen abortion.     

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)     

Microsporogenesis in a normal line and in the ogu cytoplasmic male-sterile line of Brassica napus

Summary In the ogu cytoplasmic male-sterile (CMS) line of Brassica napus, stamen morphology was influenced by temperature conditions. Under a high temperature regime (27° C/23° C; day/ night) CMS stamens had a near-normal morphology, but microsporogenesis proceeded to a maximum of the microspore stage. However, compared to the normal stamens, the occurrence of sporopollenin-like deposits in the tapetum and deposition of exine on the microspores was sparse. Also, the tapetal cells of the CMS line were often highly vacuolate and failed to degenerate at the same stage as the normal. Ultrastructural changes in the mitochondrial matrix and cristae plus dilation of the endoplasmic reticulum, which occurred during development in sporogenous tissues of the normal line, were often lacking or mistimed in the mutant. Due to extensive variation, even between adjacent locules, the cytological differences between the normal and CMS anthers cannot be ascribed as the cause of male sterility in the ogu CMS line of B. napus, rather they may be the consequence of it.     

Expression of ASK1-like genes in arrested stamens of female Silene latifolia plants

Silene latifolia is a dioecious plant in which sex is determined by heteromorphic sex chromosomes. In female plants, stamen development is arrested before microspore mother cells are formed. In this study, we isolated four cDNAs (SlSKP1-1 to 4) encoding ASK1-like protein as expression markers to reveal when expression levels are reduced in arrested stamens of female flowers. Expression patterns of the SlSKP1 genes were analyzed by in-situ hybridization. We use the flower development classification of Grant et al. (in Plant J 6:471–480, 1994). SlSKP1 genes were highly expressed in primary parietal cells and primary sporogenous cells that develop into microspore mother cells in male flowers. Expression levels started to be reduced in the external stamens of the female flowers when stamen development was arrested at stage 7. Although microspore mother cells could not be developed in female flowers and SlSKP1 expression may be unnecessary in arrested stamens, SlSKP1 genes were still expressed in sporogenous cells of degenerated stamens at stage 8. Parietal cells stopped differentiating earlier than sporogenous cells in arrested stamens. These results suggest that not all types of cell are arrested simultaneously at a particular stage of stamen development during stamen suppression of S. latifolia.     

Development of Sexual Organs in Taihangia rupestris--Different Temperature Requirments for Both Sexual Organ Development in a Bisexual Flower

Optimal environmental temperature for differentiation and development of stamen and pistil have been tested cultured in vitro flowers of Taihangia rupestris. The optimal temperatures for stamen development was quite different from that for pistil development. The most suitable temperature scope for stamen development was on the low verge of 1 ～ 6 'C. Temperature over 18 ℃ led to abortion of all stamens. The temperature scope appropriate for pistil development was near to 6～26 ℃. That below 3 ℃ brought about cessation of most pistil development at early stage. The course of stamen abortion induced by high temperature started from the primordium stage in the stamen development and once this occurred it became irreversible despite of lowering the temperature. It is concluded that the temperature for unisexual male flower, unisexual female flowers and bisexual flowers development should be contralled at 1～3 ℃, 6～26 ℃ and 6～12 ℃ respectively. The results of the present experiment may open up a new train of thoughts for studies on the control of sexual organ development and male sterility.     

Functional and evolutionary analyses of Primulina heterotricha CYC1C gene in tobacco and Arabidopsis transformation systems

In Asterids, specific expression of CYC-like genes in the corresponding regions promotes or reduces dorsal petal growth and aborts stamen development. In Rosids, however, the reduced or enlarged dorsal petals are not accompanied by the abortion of stamens, which implies that the function of CYC-like genes in regulating petal growth and stamen development might be independently recruited. To address this, we investigated the function of the PhCYC1C gene in Primulina heterotricha Y. Dong & Y. Z. Wang on petal growth and stamen development by overexpressing it in two different transformation systems, that is, Arabidopsisbelonging to Rosids and tobacco located in Asterids. The results showed that overexpression of PhCYC1C reduced petal sizes in both tobacco and Arabidopsistransgenic plants mainly by repressing cell expansion, indicating its conserved function in determining petal growth between Asterids and Rosids. However, the fertility of both tobacco and Arabidopsis stamens was not affected at all. Given that strong expression signals of PhCYC1C are detected in both tobacco andArabidopsis stamens and CYC-like genes actually function to repress stamen development in Lamiales, we suggest that the CYC-like gene-associated regulatory network for controlling stamen development might have not established in Rosids as well as in early evolution of Asterids, but evolved as Asterids proceeded further. Our results provide valuable information on the conservation of CYC-like genes' function in controlling corolla asymmetry and the divergence of their function in determining stamen abortion in angiosperms.     

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。     

Stamen dimorphism in Rhododendron ferrugineum (Ericaceae): development and function

The function of stamen dimorphism in the breeding system of the alpine shrub Rhododendron ferrugineum was studied in two populations in the French Alps. This species has pentameric flowers with two whorls of stamens: an inner whorl of five long stamens and an outer whorl of short stamens. We studied the development of stamens from buds to mature flowers (measurement of the filament, anther, and style lengths at five successive phenological stages) and compared the size and position of reproductive organs at maturity in control and partially emasculated flowers (removal of long-level stamens) to determine whether the presence of long-level stamens constitutes a constraint for the development of the short-level ones. Stamen dimorphism can be observed early in stamen development, from the bud stage of the year prior to flowering. At this early stage, meiosis had already occurred. Emasculation of the long-level stamens induced the short-level ones to grow longer than in normal conditions. We also performed seven pollination treatments on ten randomly chosen individuals in each population, and the number of seeds following each treatment was recorded. Results from these treatments showed that R. ferrugineum produced spontaneous selfed seeds in the absence of pollinators. However, no seed was produced when short-level stamens were emasculated and pollinators excluded, suggesting that long-level stamens are not responsible for selfing in the absence of pollinators and that reproductive assurance is promoted by short-level stamens.     

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。     

长春蒲公英雄性不育株形态学观察与败育细胞学研究

在长春蒲公英(Taraxacum junpeianum Kitam.)株群中发现雄性不育现象,为研究其败育机理及特点,探寻其不育基因,采用形态观察法、石蜡切片技术和染色体压片法,对长春蒲公英野生型及其雄性不育株的花药发育过程和花粉母细胞减数分裂过程进行了观察。结果表明:(1)长春蒲公英雄性不育株花药中部发红、干瘪、无花粉散出。与野生型比较,雄性不育株雄蕊更短,子房更窄,种子形态更加狭长;(2)长春蒲公英雄性不育株败育时期为四分体到单核小孢子前期,败育方式为小孢子自身异常发育,绒毡层异常分解,互相粘连败育;(3)长春蒲公英雄性不育株花粉母细胞减数分裂二分体时期出现落后微核,随后产生极少四分体,并且四分体产生大量染色体桥,小孢子营养物质流失,彻底败育。因此,长春蒲公英雄性不育株败育彻底、稳定,并且有种的特点。小孢子自身异常发育和绒毡层异常分解是导致败育的主要原因。     

MORPHOGENESIS OF THE STAMENLESS-2 MUTANT IN TOMATO. I. COMPARATIVE DESCRIPTION OF THE FLOWERS AND ONTOGENY OF STAMENS IN THE NORMAL AND MUTANT PLANTS

The single gene recessive mutant stamenless-2 (sl₂/sl₂) differs phenotypically from the normal (+/+) only in the stamen structure. Stamens of the mutant plants were laterally free, twisted, shorter, paler in color, possessed abnormal pollen, and bore naked external ovules (E.O.) on the adaxial surface near the junction of anther and filament. Mutant plants grown in the field during summer produced flowers in which a number of carpel-like organs (‘carpelloid stamens‘) with few or no E.O. replaced the stamens. On the other hand, plants grown in the greenhouse during winter possessed flowers with greater number of yellow and pubescent stamens and many E.O. Study of stamen ontogeny revealed that at initiation (up to 100 μ in length) stamen primordia of normal and mutant plants resembled each other. Thereafter the development of stamens in the two genotypes could be distinguished.     

Regulation of sex determination in maize

Maize develops separate male and female flowers in different locations on a single plant. Male flowers develop at the tip of the shoot in the tassel, and female flowers develop on the ears, which terminate short branches. The development of male flowers in tassels and female flowers in ears is the result of selective abortion of pistils or stamens, respectively, in developing florets. Genetic analysis has shown that stamen abortion and pistil abortion are under the control of two different genetic pathways. Local levels of the plant hormone gibberellic acid determine whether or not stamens are suppressed. Pistil abortion is under the regulation of the tassel seed genes, one of which has been shown to encode a short-chain alcohol dehydrogenase. The tassel seed genes play a role in regulating the fate of inflorescence meristems as well as pistil primordium fate.     

Temperature-induced modifications in the surface features of stamens of a tomato mutant: An SEM study

Summary Stamenless-2 (sl₂/sl₂) is a temperature-sensitive mutant of tomato (Lycopersicon esculentum) which exhibits altered stamen development under different temperatures (Sawhney 1983). By using scanning electron microscopy, this study was conducted to investigate the differentiation of surface features of mutant and normal stamens grown under different temperatures, with the view to further determine the role of temperature in gene expression in stamen development. Mutant stamens grown under intermediate temperatures (23 °C day/18 °C night) differed from the normal in hair production, the shape of epidermal cells and in the pattern of cuticular thickenings. Under low temperatures (18 °C day/15 °C night), all surface features of mutant stamens closely resembled the normal, whereas under high temperatures (28 °C day/23 °C night), the patterns and types of hairs, epidermal cells, stomata, and cuticular thickenings on mutant stamens were similar to that of a gynoecium. The staminal features of normal stamens were not affected by different temperatures. This study shows that the expression of the sl₂/sl₂ allele is influenced by temperature conditions to the extent that the pattern of cellular differentiation characteristic of either the stamens or the carpels can be induced in mutant stamens.     

月季花瓣数量遗传分析

以‘窄叶藤本月季花’( Rosa chinensis ‘Zhaiye Tengben Yuejihua’)ב月月粉’( R. chinensis ‘Old Blush’)杂交群体为材料, 分析其花瓣数量的分离特点, 对单瓣花与重瓣花的花芽分化过程进行观察, 并对花瓣、雄蕊及瓣化雄蕊进行表皮细胞超微结构的观察.结果显示...     

柽柳大、小孢子发生和雌、雄配子体发育的观察

利用常规石蜡制片技术,对柽柳(Tamarix chinensis Lour.)的大、小孢子发生及雌、雄配子体发育过程进行了观察。主要结果如下:(1)花药壁由五层细胞组成,从外向内分别为表皮、药室内壁,两层中层和绒毡层。药壁的发育属于基本型。绒毡层为分泌型。(2)孢原细胞为多孢原起源。小孢子母细胞减数分裂过程中的胞质分裂为连续型,形成的四分孢子为四面体型;同一药室的小孢子母细胞减数分裂几乎完全同步。(3)成熟花粉粒为2细胞型,具3个萌发孔。(4)柽柳为三心皮构成的单室复子房,每子房具有10~20个胚珠,基底胎座,胚珠为双珠被、厚珠心、倒生型。大孢子母细胞减数分裂形成1+3排列的4个大孢子, 4个大孢子全部参与胚囊的形成。(5)胚囊发育为贝母型,反足细胞在胚囊成熟时充分发育。(6)同一朵花中,前期雄蕊的发育早于雌蕊的发育,后期当花粉成熟时,雌配子体也达到成熟,雌雄蕊发育趋于同步。     

ORGAN INITIATION AND THE DEVELOPMENT OF UNISEXUAL FLOWERS IN THE TASSEL AND EAR OF ZEA MAYS

The development of the unisexual male and female flowers of Zea mays from bisexual initials in both tassels and ears has been reinvestigated with SEM and TEM. The early stages of spikelet branch primordia, spikelet initiation, and early flower development are similar in both flowers, though differences in rates of growth of glumes, lemmas, and palea were detected. In both tassel and ear flowers, a pair of stamens arises opposite the lemmas and a third stamen initiates later at right angles to the first pair but from a point on the meristem below its insertion. Gynoecia develop on both tassel and ear flowers first as a ridge which overgrows the apical meristem giving rise to the stylar canal and the elongate silk. Male flowers arise in the tassel through selective vacuolation and abortion of the cells of the early gynoecium. The single female flower in each ear spikelet arises through the vacuolation and abortion of stamens in the upper flower and the repression of growth of and the eventual regression of the lower flower in each spikelet. The significance of these selective organ abortions for practical applications is discussed.     

山鸡椒雄花花芽发育形态解剖特征观察

采用体视显微镜、扫描电镜和石蜡切片技术对山鸡椒（Litsea cubeba（Lour.） Pers.）雄花花芽分化发育的外部形态和内部解剖结构进行了观察研究。结果显示:（1）山鸡椒雄花花芽分化发生可分为5个时期,即未分化期、花序原基分化期、苞片原基分化期、花原基分化期和花器官分化期,其中花器官分化期又可细分为花被原基分化期、雄蕊原基分化期和雌蕊原基分化期;各相邻分化时期存在一定重叠现象;花期从翌年1月上旬至3月下旬。（2）雄花成熟结构中具有独特的雄蕊蜜腺,蜜腺绿色且形态不规则,着生于内轮雄蕊基部,分布于花丝两侧,夹在内外轮雄蕊的花丝之间,与内轮花丝紧密相连。（3）雄蕊花药四室,花药壁发育属于基本型;腺质绒毡层;小孢子母细胞减数分裂过程中胞质分裂属于连续型;成熟花粉为2-细胞花粉粒;成熟花粉粒外壁刺突较多,刺突基部膨大,外壁露出部分粗糙,无薄壁区,有少数小穿孔。（4）山鸡椒雄花中绝大多数雌蕊发育至腹缝线卷合形成子房室时停止,柱头发育不良或者败育,花柱缩短或缺失,不能受精,直到开花结束,即发生退化。本研究明确了山鸡椒雄花花芽发育发生各个阶段时间、形态变化特点及外部形态变化特征,山鸡椒小孢子发生、雄配子体发育至散粉期变化特点和规律以及雄花中退化雌蕊发育的进程,可为山鸡椒优良品种选育、调控花期和提高结实率提供一定的参考。     

ROX1, a gene induced by rolB, is involved in procambial cell proliferation and xylem differentiation in tobacco stamen

The Agrobacterium rhizogenes oncogene rolB mimics the effects of auxin in that it increases the sensitivity of transformed cells to this hormone. Here we isolated a tobacco gene, ROX1, acting downstream of rolB. We show that plants with reduced levels of ROX1 mRNA, due to the expression of a 35S-driven ROX1-antisense construct, have flowers with stamens and pistils longer than normal because of an increased number of cells. Localized expression of rolB in anthers results in overexpression of ROX1 and reduced growth of stamens, due to a reduced number of cells. In addition, the longer stamens of antisense plants show a delayed xylem differentiation in the lateral bundles, primarily of the junction region between anther and filament, while the shorter stamens of ROX1-overexpressing plants show a precocious differentiation of xylem cells in the same tissues. Expression of ROX1 in stamens peaks at early stages of stamen growth, and ROX1 mRNA is localized mostly in anther procambial cells. The sequence of ROX1 shares a conserved element with a number of plant genes, including TED3, which is involved in xylem differentiation. These results point to a role of ROX1 in the balance between proliferation of procambial cells and xylem differentiation during stamen development.     

光果甘草二体雄蕊的发育及其适应意义

甘草属(Glycyrrhiza)植物具“9 + 1”二体雄蕊, 其中9枚合生雄蕊的上部花丝分离, 分离的花丝在发育过程中存在由早期长、短交错的二组排列方式转变为后期以雄蕊管最长的1枚雄蕊为中心向两边渐次缩短的倒“V”形排列。为了解这种雄蕊发育动态、分化现象及发育成熟后的适应意义, 该文以光果甘草(G. glabra)为实验材料, 比较了雄蕊发育过程中的形态变化、成熟花粉的理化性质及在传粉中的作用。结果显示: 雄蕊发育早期长、短两组雄蕊在花药大小与形状上存在分化, 但后期伴随着花丝的快速生长与花粉的成熟、散出, 花药大小与形状趋于一致; 花粉组织化学成分及授粉成功率无差异, 但成熟花粉的数量和花粉活力存在差异; 去雄处理虽然使访花者在一天内的两个访花高峰期的访花频次降低, 但结实率高于自然对照, 说明以异交为主的花去除雄蕊后, 降低了雌、雄蕊间的功能干扰, 提高了传粉昆虫的授粉率; 发育早期长、短交错排列的二组雄蕊到成熟期时发生的倒“V”形排列的转变, 使不同数量与活性的花粉分布在花内不同空间, 最大化接触访花者, 实现了资源节约, 提高了雄蕊的雄性适合度, 即在有限的空间内用最节约的雄性资源投入、使传粉空间与传粉几率最大化的方式, 来提高雄性功能。