红花胚珠和雌配子体发育

用石蜡切片法研究了红花的大孢子发生和雌配子体发育过程,得到以下结果：（１）胚珠发育为薄珠心类型,倒生胚珠,具单珠被。（２）胚囊发育蓼型。（３）有珠被绒毛层,珠被绒毡层起始于大孢子母细胞时期,单核胚囊阶段高度发育,受精后从合点端逐渐退化。珠孔塞细胞呈毛状。     

ScRALF3, a secreted RALF‐like peptide involved in cell–cell communication between the sporophyte and the female gametophyte in a solanaceous species

Small peptides have been shown to regulate numerous aspects of plant development through cell–cell communication. These signaling events are particularly important during reproduction, regulating gamete development and embryogenesis. Rapid alkalinization factor (RALF)‐like genes, a large gene family that encodes secreted peptides, have specific or ubiquitous expression patterns. Previously, five RALF‐like genes with potential involvement during reproduction were isolated from Solanum chacoense. Here, we show that ScRALF3 is an important peptide regulator of female gametophyte development. Its expression, which is auxin‐inducible, is strictly regulated before and after fertilization. Down‐regulation of ScRALF3 expression by RNA interference leads to the production of smaller fruits that produce fewer seeds, due to improper development of the embryo sacs. Defects include loss of embryo sac nuclei polarization, as well as an increase in asynchronous division, accounting for cellular dysfunctions and premature embryo sac development arrest during megagametogenesis. ScRALF3 is expressed in the sporophytic tissue surrounding the embryo sac, the integument and the nucellus, as revealed by in situ hybridization and GUS staining. As expected for a secreted peptide, fluorescence from an ScRALF3–GFP fusion construct is detected throughout the secretory pathway. Therefore, the ScRALF3 secreted peptide may be directly involved in the regulation of multiple aspects of cell–cell communication between the female gametophyte and its surrounding sporophytic tissue during ovule development.     

Evidence for ovarian self-incompatibility as a cause of self-sterility in the relictual woody angiosperm,Pseudowintera axillaris (Winteraceae)

Species within the genus Pseudowintera exhibit high rates of self-sterility. Self-sterility in the genus has been previously posited-but not confirmed-to be the result of late-acting ovarian self-incompatibility (OSI) functioning within nucellar tissue of the ovule to prevent self pollen tubes from entering the embryo sac. Structural and functional aspects of pollen-carpel interactions and early seed development following cross- and self-pollination were investigated in P. axillaris to determine the site, timing and possible mechanisms of self-sterility. No significant differences were observed between pollen tube growth, ovule penetration and double fertilization following cross- and self-pollination. Pollen tubes exhibited phasic growth in an extracellular matrix composed of proteins and carbohydrates, as well as arabinogalactans/arabinogalactan proteins. A uniform failure in embryo sac development prior to division of the zygote was apparent within 15 d following double fertilization by self gametes. Results indicate that SI mechanisms in P. axillaris do not prevent double fertilization from occurring. Instead, mechanisms of self-sterility affect post-zygotic development of the embryo sac. Although self-sterility may be attributed to inbreeding depression, given the post-zygotic nature of failure in embryo sac development, the possibility of late-acting OSI is discussed.     

The female gametophyte and the endosperm control cell proliferation and differentiation of the seed coat in Arabidopsis

Double fertilization of the female gametophyte produces the endosperm and the embryo enclosed in the maternal seed coat. Proper seed communication necessitates exchanges of signals between the zygotic and maternal components of the seed. However, the nature of these interactions remains largely unknown. We show that double fertilization of the Arabidopsis thaliana female gametophyte rapidly triggers sustained cell proliferation in the seed coat. Cell proliferation and differentiation of the seed coat occur in autonomous seeds produced in the absence of fertilization of the multicopy suppressor of ira1 (msi1) mutant. As msi1 autonomous seeds mostly contain autonomous endosperm, our results indicate that the developing endosperm is sufficient to enhance cell proliferation and differentiation in the seed coat. We analyze the effect of autonomous proliferation in the retinoblastoma-related1 (rbr1) female gametophyte on seed coat development. In contrast with msi1, supernumerary nuclei in rbr1 female gametophytes originate mainly from the endosperm precursor lineage but do not express an endosperm fate marker. In addition, defects of the rbr1 female gametophyte also reduce cell proliferation in the ovule integuments before fertilization and prevent further differentiation of the seed coat. Our data suggest that coordinated development of the seed components relies on interactions before fertilization between the female gametophyte and the surrounding maternal ovule integuments and after fertilization between the endosperm and the seed coat.     

Ovule,Female and Male Gametophyte and Fertilization of Kingdonia uniflora Balfour F. et W. W. Smith

The structure of ovule, female and male gametophyte, double fertilization and the distrubution of starch grains during the fertilization have been studied. The main results are as follows: ( 1 ) Ovule The ovule is anatropous, unitegmic and tenuinucellate. The nucetlus appears cylindric, since megaspores and embryo sac development, its internal cells of nucellus become disorganized, so that only a single layer of epidermal cells remains toward the side of the micropyle, On the other hand, the integument is not as long as nucellus, as a result micropyle is not formed. And no vascular bundle is found in the integument. (2) Female gametophyte The mature embryo sac is slender and is composed of an egg cell, two synergids, a central cell and three antipodal cells. The egg cell is situated slightly away from the tip of embryo sac. Some of them contain starch grains. Synergids occupy the tip of embryo sac. Its wall at micropylar region appears irregular in thickenes and irregular in ingrowths to form the filiform apparatus. The centrateell is very large, and strongly vacuolated Two polar nuclei come to contact closely with each other, but not fuse, or to fuse into a large secondary nucleus before fertilization. The polar nuclei or the secondary nucleus are usually situated at the middle-lower position of the central cell or nearer to the chalazal end above the antipodal cell. It is different from egg cell, no starch grains are found here. In most embryo sacs three antipodal cells are found. They are not as large as those in other plants of Ranunculaceae. But six antipodal cells or the antipodal cell with two nuclei may rarely be found. Like synergid, the wall of them appears not only irregularly thickened, but clearly with irregular ingrowths. In a few antipodal cells the starch garins are usually found near the nucleus. By the end of fertilization, antipodal cells become disintegrated. (3) Male gametophyte Most pollen grains are two-celled when shedding, and rich in starch grains. A few of them contain single nucleus or three-celled. (4) The double fertilization The fertilization of Kingdonia unifiora Balfour f. et W, W. Smith is wholly similar to some plants of Ranunculaceae studied. First, the pollen tube penetrates a degenerating synergid. And the pollen tube discharges its contents with two sperm nuclei into the degenerating synergid cell. One of the two sperms fuses with the nucleus of the egg, and the other fuses with two polar nuclei or the secondary nucleus of the central cell. If one sperm nucleus at first fuses with one of the polar nuclei, and then the fertilized polar nuclei again fuses with other polar nucleus. Secondly, the fertilization of the polar nuclei or the secondary nuclei completes earlier than that of the egg. The primary endosperm nucleus begins to divide earlier than the zygote. It seems that one of the sperm nuclei come to contact with egg nucleus, the other has already fused with polar nuclei or the secondary nucleus. The zygote with a single nucleolus appears until the endosperm with 16–20 cell. Thirdly, before and after fertilization there are one to some small nucleoli in egg nucleus and polar nuclei or secondary nucleus. However they increase in quantity from the beginning of the fusion of male nucleis. These nucleoli quite differ from male nucleoli by their small size, and most of them disappear at the end of fertilization. It may be concluded that the small nucleoli increase in quantity is related to the fusion of male and female nuclei. In the duration of fertilization, in ovule starch distribution is in the basal region of integument. But in embryo sac, onlysome egg cells, or zygotes contain starch grains, a part of which was brought in by pollen tube. Sometimes the starch grains are found in some synergids and antipodal cells. No starch grains are found in the central cell.     

New insights into the functional roles of reactive oxygen species during embryo sac development and fertilization in Arabidopsis thaliana

Previously considered as toxic by-products of aerobic metabolism, reactive oxygen species (ROS) are emerging as essential signaling molecules in eukaryotes. Recent evidence showed that maintenance of ROS homeostasis during female gametophyte development is crucial for embryo sac patterning and fertilization. Although ROS are exclusively detected in the central cell of mature embryo sacs, the study of mutants deficient in ROS homeostasis suggests that controlled oxidative bursts might take place earlier during gametophyte development. Also, a ROS burst that depends on pollination takes place inside the embryo sac. This oxidative response might be required for pollen tube growth arrest and for sperm cell release. In this mini-review, we will focus on new insights into the role of ROS during female gametophyte development and fertilization. Special focus will be made on the mitochondrial Mn-Superoxide dismutase (MSD1), which has been recently reported to be essential for maintaining ROS homeostasis during embryo sac formation.     

羊草大、小孢子发生与雌、雄配子体发育的观察

利用常规石蜡制片技术研究了羊草大、小孢子发生及雌、雄配子体发育过程。主要结果是：(1)花药壁由4层结构组成,最外层为表皮,其次为药室内壁,1层中层,最内层绒毡层为分泌型;(2)小孢子母细胞减数分裂过程中的胞质分裂为连续型,四分孢子为左右对称型;(3)成熟花粉粒为3细胞型,具单萌发孔;(4)羊草为单子房、单胚珠,双珠被、薄珠心、倒生型胚珠,大孢子母细胞减数分裂形成线型或T型排列的4个大孢子;合点端大孢子具功能;(5)具有双孢原,双大孢子母细胞、双大孢子四分体和双胚囊的情况;(6)胚囊发育为蓼型,反足细胞经无丝分裂形成4～6个细胞的反足细胞群;(7)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。     

五唇兰雌配子体发育和胚胎发生的研究

五唇兰的胚珠倒生型,具薄珠心,两层珠被。胚囊发育为双孢子葱型,成熟胚囊８核。从传粉到受精约５０ｄ,正常双受精。胚具５－６细胞的胚柄,种子成熟时胚柄及胚乳核消失,成熟种子只具单层细胞的种皮和一个未分化的珠珠形胚。     

The MADS box genes SEEDSTICK and ARABIDOPSIS Bsister play a maternal role in fertilization and seed development

The haploid generation of flowering plants develops within the sporophytic tissues of the ovule. After fertilization, the maternal seed coat develops in a coordinated manner with formation of the embryo and endosperm. In the arabidopsis bsister (abs) mutant, the endothelium, which is the most inner cell layer of the integuments that surround the haploid embryo sac, does not accumulate proanthocyanidins and the cells have an abnormal morphology. However, fertility is not affected in abs single mutants. SEEDSTICK regulates ovule identity redundantly with SHATTERPROOF 1 (SHP1) and SHP2 while a role in the control of fertility was not reported previously. Here we describe the characterization of the abs stk double mutant. This double mutant develops very few seeds due to both a reduced number of fertilized ovules and seed abortions later during development. Morphological analysis revealed a total absence of endothelium in this double mutant. Additionally, massive starch accumulation was observed in the embryo sac. The phenotype of the abs stk double mutant highlights the importance of the maternal-derived tissues, particularly the endothelium, for the development of the next generation.     

Early Events in the Penetration of the Embryo Sac in Torenia fournieri(Lind.)

At maturity, Torenia fournieri(Lind.) has an embryo sac whichprotrudes through the micropyle placing the synergids, egg celland part of the central cell within the ovary locule adjacentto the placenta. The present study utilized this unique attributein combination with confocal and light microscopy to characterizethe timing and associated structural changes during pollinationevents leading to double fertilization. The observation of spermnuclei in living gametophyte tissue is an important advancein the identification, in real time, of stages leading to fertilizationin angiosperms. A continuum of fertilization occurred between12 and 16 h after pollination (hap), with peak frequency ofegg and sperm fusion at 14 hap (43%). Movement of the spermcells through the degenerated synergid took several hours andfusion between sperm and their respective female nuclei occurredsimultaneously. Changes in embryo sac structure were also documented.Cell walls in the region between the synergids and egg cellwere poorly developed prior to pollen tube penetration. Thickenedcell walls were observed around the periphery of the synergidsand egg cell following pollination, and in the central cellwhere it lay within the body of the ovule. Starch was observedin the cells of the embryo sac, although the number and distributionof granules varied before and after pollination. These temporaland spatial observations of the embryo sac inTorenia fournieriprovide a basis for further research to determine control mechanismsoperating during specific double fertilization events in angiosperms.Copyright 2000 Annals of Botany Company Double fertilization, embryo sac, sperm nuclei, Hoechst, Torenia fournieri     

小蓬草的胚胎学研究

对小蓬草(Conyza canadensis)大小孢子发生、雌雄配子体形成、受精、胚及胚乳发育过程进行了研究,主要结果如下:花药四室,药壁由表皮、药室内壁、中层和绒毡层组成.表皮退化;药室内壁宿存,细胞柱状伸长,纤维状加厚;中层细胞退化较早,在小孢子母细胞减数分裂开始时仅存残迹;绒毡层于小孢子母细胞减数第一次分裂前期开始原位变形退化,属于腺质型绒毡层;小孢子母细胞减数分裂为同时型,四分体的排列方式主要为四面体形和左右对称形;成熟花粉粒多为3细-胞花粉粒,偶见2细-胞花粉粒.子房下位,2心皮,1室,单胚珠,基生胎座;单珠被,薄珠心,倒生胚珠,具发达的珠被绒毡层.珠心表皮下分化出大孢子孢原细胞,孢原细胞直接发育为大孢子母细胞,大孢子母细胞减数分裂形成4个大孢子直线形排列,仅合点端的大孢子发育成功能大孢子母细胞,胚囊发育为蓼型.两个极核在受精前融合为次生核,珠孔受精.胚乳发育属于核型,胚胎发育为紫菀型;具胚乳吸器.     

Attraction and transport of male gametes for fertilization

 Two capabilities are critical in attracting and transporting male gametes for fertilization: (1) the pollen tube must locate, enter and discharge its contents at the correct site within the female gametophyte, and (2) once inside the embryo sac, the non-motile male gametes must be transported to the egg and the central cells for double fertilization. This review summarizes current information about evidence for communication between embryo sac and pollen tube and the means by which the non-motile male gametes are transported from the aperture of the pollen tube to the site of gamete fusion. Received: 6 June 1996 / Revision accepted: 9 July 1996     

大花君子兰大小孢子发生与雌雄配子体发育的研究

该研究运用常规石蜡切片技术,对大花君子兰(Clivia miniata Regel)大、小孢子发生及雌、雄配子体发育进程进行解剖学观察分析,以探讨君子兰生殖生物学解剖特征,为君子兰种子发育和育种提供理论依据。结果表明：(1)大花君子兰花药4室,具分泌型绒毡层。(2)小孢子母细胞减数分裂的胞质分裂为连续型,小孢子四分体为左右对称型,成熟花粉为二细胞型。(3)倒生胚珠,双珠被,厚珠心和雌配子体发育为蓼型。(4)记录了雌雄配子体发育的对应关系,发现雄配子体发育趋于同步,雌配子体发育不同步。(5)开花散粉时,雌配子体尚有处于四核、八核胚囊的时期;成熟胚囊阶段,中央细胞的2个极核位于反足细胞端,反足细胞呈退化状态。具承珠盘结构。     

小蓬草的胚胎学研究

对小蓬草（Conyzacanadensis）大小孢子发生、雌雄配子体形成、受精、胚及胚乳发育过程进行了研究,主要结果如下：花药四室,药壁由表皮、药室内壁、中层和绒毡层组成。表皮退化;药室内壁宿存,细胞柱状伸长,纤维状加厚;中层细胞退化较早,在小孢子母细胞减数分裂开始时仅存残迹;绒毡层于小孢子母细胞减数第一次分裂前期开始原位变形退化,属于腺质型绒毡层;小孢子母细胞减数分裂为同时型,四分体的排列方式主要为四面体形和左右对称形;成熟花粉粒多为3-细胞花粉粒,偶见2-细胞花粉粒。子房下位,2心皮,1室,单胚珠,基生胎座;单珠被,薄珠心,倒生胚珠,具发达的珠被绒毡层。珠心表皮下分化出大孢子孢原细胞,孢原细胞直接发育为大孢子母细胞,大孢子母细胞减数分裂形成4个大孢子直线形排列,仅合点端的大孢子发育成功能大孢子母细胞,胚囊发育为蓼型。两个极核在受精前融合为次生核,珠孔受精。胚乳发育属于核型,胚胎发育为紫菀型;具胚乳吸器。     

矮沙冬青雌配子体及胚胎发育研究

矮沙冬青子房单心皮1室,边缘胎座,弯生胚珠,胚珠具双珠被、厚珠心。大孢子孢原细胞发生于珠心表皮下,大孢子母细胞减数分裂形成直线排列的四分体,合点端大孢子具功能,并按蓼型胚囊发育,雌配子体成熟于4月中旬。双受精后,胚乳发育为核型。在矮沙冬青大孢子发生、雌配子体和胚胎发育过程中未发现异常现象,因此认为矮沙冬青濒危不存在雌性生殖结构与发育过程异常的内在因素。     

鹅掌楸雌配子体败育对生殖的影响

胚珠和雌配子体败育是限制鹅掌楸生殖成功的一个重要因素。中国东部和西部鹅掌楸种群在雌配子体发育的各阶段上的败育程度有差异,以西部种群的发育较好。西部分布区较合适的生境促进了胚囊的发育,一定温度和湿度的环境可以活化珠心细胞输送营养物质供给雌配子体发育,提高受精和结籽的能力     

八角莲大孢子发生和雌配子体形成

首次报道了八角莲(Dysosma versipellis (Hance)M.cheng)大孢子发生和雌配子体形成的过程.结果:双珠被,多为厚珠心胚珠,少数为假厚珠心,胚珠多为横生,少数为弯生;边缘胎座,子房一室,多胚珠,珠孔由两层珠被共同形成,呈"之"字形;多为单孢原,位于珠心表皮下:偶见2～3个孢原细胞位于珠心表皮下;大孢子母细胞有两种发生方式;直线形大孢子四分体,合点端的大孢子发育为功能大孢子,蓼型胚囊;成熟胚囊中,二个极核在受精前合并为次生核;三个反足细胞不发达,较早退化;"品"字形卵器极性明显,其中卵细胞与助细胞极性相反;助细胞发达,其丝状器在不同发育时期形态及大小不同,且具吸器功能.     

千里光配子体发育、配子形成与双受精的细胞学特征

千里光（Senecio scandens Buch.-Ham.ex D.Don）是具有良好开发前景的传统抗菌中草药。为了探讨该物种有性生殖的细胞学机制,实现在物种的遗传育种过程中的指导作用,本研究从细胞学角度观察了千里光配子体发育、配子形成和双受精作用。结果表明,在生殖核分裂为精细胞的过程中,其位置和形态呈现差异,表现为"二态"精细胞;此外,雌配子体的形成符合孢子发育模式,成熟胚囊的形态结构为卵形或梨形,成熟的卵细胞和极核位于珠孔端。根据上述结果可推断,合子在随后二分裂时表现为"基细胞-顶细胞"极性,可能与"二态"精细胞导致的合子细胞质不均一有关。此外,从植物分类学角度,千里光的"二态"精细胞,胚囊极性和合子极性可成为菊科近缘植物分类的细胞学依据。