黄檗(芸香科)的珠孔塞和珠孔的形态发育及花粉管在子房室中的生长路径

利用扫描电子显微镜术和光学显微镜术研究了黄檗(Phellodendron amurense Rupr.)的珠孔塞和珠孔的形态发育和花粉管在雌蕊中的路径.黄檗胚珠的珠孔塞起源于珠柄.随着胚珠生长,珠孔塞逐渐增大,胚珠成熟时珠孔塞变得相当大并紧密地覆盖在珠孔上.当雌花进入可传粉期时,珠孔塞的形态发生很大变化,其表面细胞径向延伸,形成柱形、半乳突或乳突细胞.受精后,珠孔塞体积变小并逐步退化.花粉管在子房室中并非一定经过珠孔塞结构.花粉管是否经过珠孔塞取决于它们进入子房室的位置.我们不支持先前研究者关于珠孔塞主要充当对花粉管生长的机械作用的观点.我们对黄檗胚珠的珠孔的形态发育研究显示,在不同的生殖时期,珠孔的结构会发生变化,在传粉时期它的结构显示不对称性.黄檗珠孔塞和珠孔的发育与雌配子体发育存在密切关系.     

珠心喙与珠孔塞

大戟属（Euphorbia）植物的胚珠普遍具有珠心喙和珠孔塞的结构。珠心喙是由珠心伸入珠孔之中形成的喙状结构。珠孔塞是由胎座细胞衍生而形成的毛状细胞（图1）。实验表明,大戟属植物珠心喙的长短与珠孔塞的发达与否密切相关。珠心喙长并伸出珠孔外者,其珠孔塞不明显或无（图2）。珠心喙短不伸出珠孔外者珠孔塞发达,珠孔塞的毛状细胞进入珠孔而与珠心喙相接触（图3．4）。珠心喙和珠孔塞的存在与引导花粉管进入胚囊有关。珠心喙与珠孔塞@齐淑艳$沈阳大学师范学院生物系!110015     

Fuerte和Hass品种油梨的花粉管生长、受精与早期胚和胚乳发育的光学显微镜研究

用光学显微镜对Fuerte和Hass品种油梨的花粉管生长、受精与早期胚和胚乳发育进行了研究。授粉后24小时,花粉管穿入胚珠。当花粉管到达子房时,花粉管贴着子房内壁表面生长,然后沿着珠柄,穿过内珠被形成的珠孔,在珠心顶端的乳突细胞之间生长。它经过一个助细胞进入胚囊。授粉后48小时在胚囊中可见到精核,精核与极核融合后,精卵才融合。胚乳核先分裂,接着细胞壁形成。授粉后5—6天,合子第一次分裂。Fuerte品种授粉后1—2天的胚珠中,虽然在珠被或珠心处经常看到花粉管,但是只有不到20%的胚囊有花粉管进入。在Hass品种中,有60%的胚囊有花粉管穿入。可以认定,Fuerte品种之所以低产可能与花粉管很少进入胚囊有一定的关系。     

陆地棉珠孔的结构及花粉管在其中的生长途径

在大多数被子植物中,花粉管经珠孔进入胚珠进而实现受精,但珠孔结构及花粉管在其中的生长途径是植物受精生物学研究中较薄弱的一个环节。迄今仅在向日葵中从超微结构水平上作了详细的研究［１］,因而需要扩大研究的对象。陆地棉花粉管生长所经雌蕊组织中,关于柱头、花柱［２,３］、珠心［４,５］和助细胞［６］的超微结构已有较详细的研究,但涉及珠孔这一环节,只有简单的描述［７］。本文报道我们在这方面的研究结果。材料和方法取陆地棉（ＧｏｓｓｙｐｉｕｍｈｉｒｓｕｔｕｍＬ．）授粉前和授粉后２２小时的胚珠,切除其合点端部分…     

油橄榄授粉受精和胚胎发育过程观察

该研究以油橄榄“鄂植-8”为材料,应用压片荧光观察花粉在柱头和花柱中的萌发及生长情况,采用石蜡制片法观察油橄榄的胚珠结构特点和授粉受精过程。结果表明：油橄榄为1子房2心室4胚珠,珠心较发达,由多层细胞构成,属于厚珠心椭圆型胚珠,胚珠直生;花开时花粉粒落到柱头上立刻萌发,随后花粉管在花粉通道中生长,再后进入子房经子房内表面,大部分花粉管不能到达胚珠,仅少数沿胎座生长经珠柄进入珠孔,释放2个精子,2个精核分别进入卵细胞和极核,并与卵核及极核相互融合;观察到合子中雌、雄性核仁融合的过程。授粉受精各阶段经历的时间为花粉落到柱头上立刻萌发;授粉后6d左右,花粉管长入胚珠的珠孔,随后释放精子;10 d左右完成授粉受精,30 d左右形成心型胚,40 d左右胚发育几乎成熟。在进行子房石蜡切片中,共观察到612个完整子房切片,发育完整的子房为97.53％,完成授粉受精的胚珠为15.52％。油橄榄在自然授粉下,约85％的胚珠授粉受精过程不能完成,在一定程度上影响其坐果。该研究结果为油橄榄授粉受精、高效生产等提供了理论依据。     

鹤顶兰花粉管在子房中的生长途径

运用扫描电镜对鹤顶兰(Phaiustankervilliae(Aiton)Bl.)花粉管在子房内的生长途径进行了观察。结果表明:花粉管在子房中的生长途径可以分为3个阶段:(1)沿子房壁轴向生长阶段,从授粉开始至大孢子母细胞四分体时期,花粉管经过合蕊柱到达子房,经由胎座基部沿子房壁轴向生长;(2)沿子房径向生长阶段,二核胚囊之后,花粉管在胚珠之间穿梭,以径向生长为主;(3)朝珠孔定向生长阶段,胚囊成熟时,花粉管朝珠孔定向生长进入胚囊。实验结果说明花粉管的定向生长受胚珠的分子信号调控。     

向日葵柱头,花柱和珠孔中钙分布的超微细胞化学定位

用焦锑酸盐沉淀法对向日葵（ＨｅｌｉａｎｔｈｕｓａｎｎｕｕｓＬ．）授粉前后柱头、花柱和珠孔中的钙进行了超微细胞化学定位。同时还运用Ｘ射线能谱（ＥＤＸ）和波谱（ＷＤＸ）两种方法进行了Ｘ射线定性分析,证明了前法所得沉淀确系焦锑酸钙。观察表明,花粉萌发和花粉管生长所经的柱头接受面,花柱引导组织和珠孔引导组织中含钙较柱头、花柱和珠孔的其它部位明显地多。柱头乳突细胞的表面和花柱引导组织的胞间基质中、尤其胞间基质与细胞壁外层相接之处钙很密集。在珠孔外端引导组织中,以角质层为界,钙主要分布于其近珠柄侧。花粉管壁果胶质层中有相当多的钙。结合向日葵中已有的研究和其他文献,讨论了钙的分布与花粉管生长的关系     

美味猕猴桃和软枣猕猴桃种间杂交花粉管行为和早期胚胎发生的观察

美味猕猴桃（Ａｃｔｉｎｉｄｉａ ｄｅｌｉｃｉｏｓａ Ｎｏ．２６）和软枣猕猴桃（Ａ． ａｒｇｕｔａ）杂交,观察了花粉管在花柱内的行为和早期胚胎发生,结果如下： 花粉粒在柱头的乳突细胞表面萌发,在开放型的Ｖ 形花柱道内生长,生长速度比对照的缓慢, 到达胚珠珠孔处的时间延迟５０ 到６０ ｈ。当花粉管生长到花柱基部时出现波纹状弯曲;顶端膨大或尖细;部分花粉管的直径发生变化;少数花粉管生长极不规则。花粉管中胼胝质沿管壁不规则沉积, 有的不形成胼胝质塞,或整个管壁被胼胝质所覆盖。约有２６．７４％ 的胚珠受精, 并发育成种子,其中正常种子占６８．５０％ ;败育种子占３１．５０％ ,其中约有１１．４１％ 为空瘪种子;未受精胚珠占６１．４５％ 。正常种子及其胚均较对照的小。胚发育为茄型,胚乳为细胞型,合子保持休眠十几天后开始分裂形成胚,传粉后５０ｄ 见到子叶胚     

非洲狼尾草胚珠(着重胚珠附器)发育的特点

用光学显微镜对非洲狼尾草胚珠发育做了进一步观察。结果表明有如下几个特点：（１）珠孔区域的珠心细胞发生特化,膨大１伸长并进入珠孔,形成胚珠附器。（２）珠心表皮平周分裂产生周缘珠心组织。（３）珠孔由内珠被和腹侧外珠被构成。     

甘蔗雌雄蕊的解剖学研究

甘蔗雄蕊花药的壁由四层细胞构成,药室内壁在花药成熟时才发育。花粉母细胞减数分裂属连续型,四分体正常发育为小孢子,但当开花时,只有少数进一步发育为成熟的花粉粒。甘蔗雌蕊柱头是分枝的,每一柱头毛是由数个细胞质浓密的细胞,呈不规则的纵列而组成。倒生胚珠一个,着生于子房室的内侧;内、外珠被均由两层细胞构成,外珠被上侧未能生长到珠孔,内珠被的内层细胞在胚珠发育中不断增大体积。石蜡制片是以樟油为透明剂的。     

Mode of pollen-tube growth in Pistils of Myrica rubra (Myricaceae): a comparison with related families

BACKGROUND AND AIMS: It is generally known that fertilization is delayed for more than a few weeks after pollination in Fagales. Recent studies showed that, during that period, pollen tubes grew in pistils in close association with the development of the ovule in a five-step process in Casuarina (Casuarinaceae) and a four-step process in Alnus (Betulaceae). The number of pollen tubes was reduced from many to one, a fact suggesting that delayed fertilization plays a role for gametophyte selection. Myrica (Myricaceae) also shows delayed fertilization for >2 weeks after pollination, but nothing is known of how pollen tubes grow in the pistil during that period. METHODS: Pollen-tube growth and the development of the ovule in pistils was investigated by fluorescent and scanning electron microscopy and analysis of microtome sections of the pistils. KEY RESULTS: Developmental study of the pollen-tube growth in the pistil of M. rubra showed that the tip of the pollen tube was branched or lay in a zigzag pattern in the upper space of the ovarian locule or near the tip of the integument, and subsequently was swollen on the nucellar surface. Such morphological changes indicate that the pollen-tube growth was temporarily arrested before fertilization. The pollen-tube growth in M. rubra can therefore be summarized as occurring in three steps: (1) from the stigma to the ovarian locule; (2) from the ovarian locule to the nucellar surface; and (3) from the nucellar surface to the embryo sac. CONCLUSION: Myrica differs from other families in that the pollen tubes arrest their growth on the nucellar surface, probably digesting nutrient from nucellar cells. There is little information on five other families of Fagales. An extensive study is needed to better understand the diversity and function of the mode of pollen-tube growth within the order.     

Evidence for Stigmatic Self-incompatibility, Pollination Induced Ovule Enlargement and Transmitting Tissue Exudates in the Paleoherb, Saururus cernuus L. (Saururaceae)

Saururus cernuus, a species belonging to the primitive herbaceousangiosperm family Saururaceae, exhibits high rates of self-sterility.We investigated the structural and functional aspects of pollen-carpelinteractions following cross and self pollination to assessthe tissue specific site and timing of self-sterility and factorsimportant for successful cross pollen tube growth. Self-sterilitywas due to inhibition of self pollen germination at a dry stigma.Self pollination was associated with anomalous foot formation,reduced cell wall expansion and secretory activity of stigmaticpapillae, and callose production in stigmatic papillae. Followinggermination, cross compatible pollen tubes entered a solid coreof transmitting tissue and grew to the base of a short style.Entry of cross pollen tubes into the ovary was coincident withovule enlargement which placed the micropyle in the proximityof cross pollen tube tips. Ovule enlargement also occurred followingself pollination. Cross pollen tubes either entered an exudate-filledmicropyle directly from the style, or growth in the ovary waslocalized to the epidermis of the locule and outer integumentprior to entry into the micropyle. Prior to pollination, thetransmitting tract was void of secretions except for exudatein the micropyle. Growth of pollen tubes on the locule and integumentwas associated with exudate apparently arising from transmittingcells adjacent to growing pollen tubes. The present study providesthe first evidence in a primitive herbaceous species of stigmaticself-incompatibility (SI) in association with a dry stigma,pollination-induced signalling events affecting developmentof carpellary tissues, and micropylar exudates. Copyright 1999Annals of Botany Company SI evolution, dry stigma, exudates, pollen-carpel signalling, Saururaceae.     

SOME ASPECTS OF CARPEL STRUCTURE IN CALTHA PALUSTRIS L. (RANUNCULACEAE)

Caltha palustris L. carpels obtained from closed flower buds, from flowers that had just reached anthesis, and from older flowers were examined by light and electron microscopy. Trichomes located on either side of the cleft towards the base of each carpel, cells along the margins of the carpel cleft, and transfer cells along the locule lining immediately beneath the micropyle of the anatropous ovule were examined. Numerous, smooth endoplasmic reticulum cisternae and dictyosomes, the presence of material between the cell wall and cuticle, and droplets of material in the region of trichomes is evidence that the trichomes are nectaries. The cells lining the cleft and the transfer cells which have wall ingrowths along the tangential wall facing the locule may be involved in the secretion of substances for pollen tube growth.     

Pollen tube growth and guidance is regulated by POP2, an Arabidopsis gene that controls GABA levels

During angiosperm reproduction, pollen grains form a tube that navigates through female tissues to the micropyle, delivering sperm to the egg; the signals that mediate this process are poorly understood. Here, we describe a role for gamma-amino butyric acid (GABA) in pollen tube growth and guidance. In vitro, GABA stimulates pollen tube growth, although vast excesses are inhibitory. The Arabidopsis POP2 gene encodes a transaminase that degrades GABA and contributes to the formation of a gradient leading up to the micropyle. pop2 flowers accumulate GABA, and the growth of many pop2 pollen tubes is arrested, consistent with their in vitro GABA hypersensitivity. Some pop2 tubes continue to grow toward ovules, yet they are misguided, presumably because they target ectopic GABA on the ovule surface. Interestingly, wild-type tubes exhibit normal growth and guidance in pop2 pistils, perhaps by degrading excess GABA and sharpening the gradient leading to the micropyle.     

Ultrastructure of the micropyle and its relationship to pollen tube growth and synergid degeneration in sunflower

Summary Ultrastructural studies made on the micropyle of sunflower before and after pollination resulted in the following observations. (1) The micropyle is closed instead of a hole or canal. The inner epidermis of the integument on both sides of the micropyle is in close contact at the apex of the ovule. The boundary between the two sides consists of two layers of epidermal cuticle. (2) The micropyle contains a transmitting tissue. The micropyle is composed of an intercellular matrix produced by the epidermal cells of the integument. (3) The micropyle is asymmetrical, and is much wider on the side proximal to the funicle. On the funicle side the cells adjacent to the micropyle are similar to those of the transmitting tissue: they have large amounts of intercellular matrix and contain abundant dictyosomes, rough ER, and starch grains, and provide an appropriate environment for growth of the pollen tubes. The cells distal to the funicle are rich in rough ER and lipid bodies; they lack large intercellular spaces. (4) The micropyle is variable in the axial direction, i.e., it is much larger and more asymmetric at the level distal to the embryo sac than at a level close to the embryo sac. After pollination, one to four pollen tubes are seen in a micropyle. During their passage through the micropyle, most pollen tubes are restricted to the side proximal to the funicle. There is a greater tendency (81%) for the degenerate synergid to be located toward the funicle, i.e., at the same side as the pollen tube pathway. The data indicate a close relationship between micropyle organization, orientation of pollen tube growth, and synergid degeneration.     

Self-incompatibility inCrocus vernus subsp.vernus (Iridaceae)

Outcross, self- and mixed pollinations were performed inCrocus vernus subsp.vernus, a species with bicellular pollen, dry stigmas and hollow style. No differences were noted among the above pollinations concerning the germination of pollen and the growth of pollen tubes until the top of ovary. Within 45 min after pollinations 62% of pollen grains germinated. Pollen tubes penetrated the papilla cuticle extending along the papilla wall; on entry into stigmatic lobes they continued growth in the stylar secretion to ovarian locules. Here, however, self-pollen tubes failed to reach or to enter the ovule micropyle; while pollen tubes from either outcross- or mixed pollinations grew until fertilizing ovules. These observations gave evidence of a self-incompatibility system inCrocus, which appeared to be neutralized by mentor effect. The ovary as site of incompatibility response is discussed.     

DYNAMICS OF POLLEN TUBE GROWTH IN THE WILD RADISH RAPHANUS RAPHANISTRUM (BRASSICACEAE). II. MORPHOLOGY,CYTOCHEMISTRY AND ULTRASTRUCTURE OF TRANSMITTING TISSUES,AND PATH OF POLLEN TUBE GROWTH

The relative importance of prezygotic mechanisms of gametophytic competition and selection are often unclear due to an absence of observations on the gynoecium and pollen tube growth in vivo. We used LM, SEM, and TEM to study the structure of the gynoecium and the path of pollen tube growth in Raphanus raphanistrum, a sporophytically self-incompatible annual. Wild radish has a papillate stigma and a solid style. A septum, which is characteristic of cruciferous gynoecia, is present in the ovary. After germination on the stigma, pollen tubes grow in the secretion of the transmitting tract of the style. The stylar secretion stains positive for acidic polysaccharides and insoluble carbohydrates, and negative for lipids and protein. In the ovary, the transmitting tissue is contained inside the septum. The secretion in the ovary stains positive only for acidic polysaccharides. Pollen tubes travel inside the septum as they enter the ovary and must exit to the surface of this tissue before ovule fertilization can occur. Pollen tube growth on the septum tracks the intercellular junctions of the septum epidermis where the secretion leaks out through a torn cuticle. Tubes must grow across the obturator before reaching the micropyle of an ovule. The temporal pattern with which tubes growing into the ovary exit the septum can contribute to the previously observed nonrandom patterns of fertilization (Hill and Lord, 1986).     

Guidance in vitro of the pollen tube to the naked embryo sac of torenia fournieri

The precise guidance of the pollen tube to the embryo sac is critical to the successful sexual reproduction of flowering plants. We demonstrate here the guidance of the pollen tube to the embryo sac in vitro by using the naked embryo sac of Torenia fournieri, which protrudes from the micropyle of the ovule. We developed a medium for culture of both the ovule and the pollen tube of T. fournieri and cocultivated them in a thin layer of solid medium. Although pollen tubes that had germinated in vitro passed naked embryo sacs, some pollen tubes that grew semi-in vitro through a cut style arrived precisely at the site of entry into the embryo sac, namely, the filiform apparatus of the synergids. When pollen tubes were unable to enter the embryo sac, they continuously grew toward the same filiform apparatus, forming narrow coils. Pollen tubes selectively arrived at complete, unfertilized embryo sacs but did not arrive at those of heat-treated ovules or those with disrupted synergids. These results convincingly demonstrate that pollen tubes are specifically attracted to the region of the filiform apparatus of living synergids in vitro.     

Structural aspects of in vitro pollen tube growth and micropylar penetration inGasteria verrucosa (Mill.) H. Duval andLilium longiflorum Thunb.

Summary In vitro penetration of the micropyle of freshly isolatedGasteria verrucosa ovules by pollen tube was monitored on agar medium. 40–60% of the micropyles were penetrated, comparable with in vivo penetration percentages. When germinated on agar,Gasteria pollen tube elongation lasts for up to 8 h while plasma streaming continues for about 20–24 h. The generative cell divides between 7 and 20 h after germination, and after 20 h the pollen tube arrives at one of the synergids. The sperm cells arrive after 22 h. The whole process takes more time in vitro than in vivo. In fast growing pollen tubes, a pulsed telescope-like growth pattern of tube elongation is observed. The formation of pollen tube wall material precedes tube elongation and probably prevents regular enlargement of the pollen tube tip-zone. Rapid stretching of the new pollen tube wall material follows, probably due to gradually increased osmotic pressure and the use of lateral wall material below the tip. The stretching ceases when the supplies of plasma membrane and excretable wall material are exhausted. Multiple pollen tube penetration of the micropyle occurs in vitro as it does in vivo. Most pollen tube growth ceases within the micropyle but, if it continues, the pollen tubes curl. Inside the micropyle the pollen tube shows haustorial growth. At the ultrastructural level, the wall thickening of in vitro pollen tubes is quite similar to that in vivo. Before transfer of pollen tube cytoplasm a small tube penetrates one of the synergids. Sperm nuclei with condensed chromatin are observed in the pollen tube and the synergid. In vivo prometaphase nuclei are found in the most chalazal part of a synergid, against the egg cell nucleus and nucleus of the central cell at a later stage. Using media forLilium ovule culture,Gasteria ovules were kept alive for at least 6 weeks. Swelling of the ovule depends on pollen tube penetration. The conditions for fertilization to occur after in vitro ovular pollination seem to be present.