菜豆花粉发育中质体和线粒体及其DNA存在的状况——着重阐明质体遗传…

应用电镜和ＤＮＡ的ＤＡＰＩ荧光检测技术研究了菜豆小孢子／花粉发育中质体和线粒体及其ＤＮＡ存在的状况。观察表明：在小孢子分裂时质体全部分配到营养细胞中，初形成的生殖细胞已不含质体。线粒体和质体的ＤＮＡ在花粉发育中也先后降解，生殖细胞从刚形成时发育至成熟花粉时期这两种细胞器ＤＮＡ均不存在。研究结果为菜豆质体母系遗传提供了确切的细胞学证据。遗传分析的研究曾确定菜豆质体为双亲遗传，与对本研究结论不同的原因     

豌豆生殖细胞的发育—着重论述质体母系遗传的细胞学基础

超微结构的研究证明,豌豆（Ｐｉｓｕｍ ｓａｔｉｖｕｍ Ｌ．）生殖细胞自形成直至成熟花粉时期,始终存在少量质体和较多的线粒体。ＤＮＡ 荧光的观察表明,在发育早期的生殖细胞中不含细胞质ＤＮＡ 类核,但在成熟花粉的生殖细胞中有许多的类核。在花粉离体萌发过程中,随着花粉管的生长,生殖细胞中的类核逐渐降解。在花粉培养２４ ｈ 后,生殖细胞的类核全部消失。研究结果确定了豌豆质体母系遗传的细胞学基础,支持遗传分析及ＲＦＬＰ研究的结论,阐明了过去在细胞学上认为是双亲遗传的判断不正确的原因     

杜鹃精细胞的超微结构及DNA荧光观察——着重阐明质体双亲遗传的细胞学基础

迎红杜鹃 ( Rhododendron mucronulatum Turcz.)的成熟花粉为二细胞型 ,精细胞在花粉管中形成。花粉管中的两个精细胞及与营养核之间互相联结 ,形成雄性生殖单位。两个精细胞的细胞质中均含有丰富的细胞器 ,包括质体、线粒体、小泡及微管 ,内质网和高尔基体稀少。具正常结构的精细胞质体在切面上多呈环形或哑铃形 ,内膜不发达 ,基质电子密度高。线粒体为球形或棒状 ,基质电子密度较低。 DNA特异性荧光染色显示 ,生殖细胞及精细胞中均含有大量类核 ( nucleoid) ,两个精细胞中的类核数量无明显差异。结果证明了杜鹃精细胞中存在大量具 DNA的可遗传细胞器 ,为杜鹃属植物的双亲细胞质遗传方式提供了细胞学证据。     

被子植物质体遗传的细胞学研究

植物细胞质遗传涉及细胞质中含DNA的两种细胞器——质体和线粒体从亲代至子代的传递。相对来说线粒体遗传的研究远不及质体的多,这可能是线粒体这种细胞器缺乏合适的表型突变体之故。高等植物质体遗传的研究历史可追溯到本世纪初在杂交试验中对叶色遗传的非孟德尔定律的发现,Baur在马蹄纹天竺葵(Pelargonium zonale)中从叶色突变体(白化体)的杂交遗传分析,发现了双亲质体遗传;而Correns在紫茉莉(Mirabilis jalapa)中则发现了单亲母本质体遗传(见Kuroiwa)。此后,对质体基因组突变性状遗传分析的研究,大量的资料说明了在被子植物中存在双亲质体遗传和单亲母系质体遗传两种类型,而后一种占大多数,仅少数是比较有规律的为双亲质体遗传或偶尔是双亲质体遗传。几十年来应用遗传分析的方法对被子植物质体遗传的研究,着重于揭示不同植物种质体的遗传是单亲母系或是双亲质体传递,以及探索杂种核基因对质体传递方式的影响。     

杜鹃精细胞的超微结构及DNA荧光观察：着重阐明质体双亲遗传的细胞学…

迎红杜鹃（ＲｈｏｄｏｄｅｎｄｒｏｎｍｕｃｒｏｎｕｌａｔｕｎＴｕｒｃａ．）的成熟花粉为二细胞型,精细胞在花粉管中形成。花粉管中的两个精细胞及与营养核之间相联结,形成在雄性殖单位。两个精细胞的细胞质中均含有丰富的细胞器,包括质体,线粒体,小泡及微管,内质网和高尔基体稀少。     

天竺葵质体和线粒体双亲遗传的细胞学机理——雄性和雌性配子超微结构和DNA荧光的研究

天竺葵（Ｐｅｌａｒｇｏｎｉｕｍ ｈｏｒｔｏｒｕｍ Ｂａｉｌｅｙ）生殖细胞和精细胞在发育中始终存在质体和线粒体。在精细胞中,质体的体积大、数量多,具基质浓厚和在切面上多为环状的特点。线粒体在生殖细胞和精细胞中没有差异,体积较质体小得多,球形或杆状,边缘染色较深。在卵细胞中质体的含量比线粒体丰富,这两种细胞器的结构形态与精细胞的有明显的差异。细胞的质体多呈不规则的棒状和含淀粉粒。线粒体比精细胞的大２—３ 倍,许多为环状。ＤＮＡ 荧光的检测证明了在生殖细胞、精细胞和卵细胞中存在质体和线粒体类核。卵中的环状线粒体类核的形态在精细胞中是不存在的。本研究确定了雄性和雌性配子的质体和线粒体在结构形态上各具特点,可作为鉴别它们的标记,从合子中查明雄性质体和线粒体是否传递,以及在胚胎发育的早期雌雄亲本来源的细胞器的动态     

天竺葵质体和线粒体双亲遗传的细胞学机理——雄性和雌性配子微结构…

天竺葵生殖细胞和精细胞在发育中,始终存在质体和线粒体。在精细胞中,质体的体积大、数量多、具基质浓厚和切面上多为呈现的特点。线粒体在生殖细胞和精细胞中没有差异,体积较质体小得多,球形或杆状,边缘染色较深。在卵细胞中质体的含量比线粒体丰富,这两种细胞器的结构形态与精细胞的有明显的差异。细胞的质体多呈不规则的棒状和含淀粉粒。线粒体比精细胞的大２－３倍,许多为环状。ＤＮＡ荧光的检测证明了在生殖细胞、精细胞     

牵牛属质体DNA的父系遗传

利用限制性片段长度多态性（ＲＦＬＰ）技术研究了牵牛属（Ｐｈａｒｂｉｔｉｓ）植物质体ＤＮＡ 的遗传方式。结果表明,在牵牛（Ｐ． ｎｉｌ）×大花牵牛（Ｐ． ｌｉｍｂａｔａ）和大花牵牛×牵牛中,质体ＤＮＡ 为父系遗传。在大花牵牛×牵牛中,质体ＤＮＡ还有可能为双亲遗传。研究证明牵牛属为被子植物中具有质体父系遗传方式的第三个属。牵牛属质体父系遗传机制尚不清楚,作者认为母系质体及其ＤＮＡ 在受精后的释稀、排除和／或降解可能是质体父系遗传的基础     

玉竹雄配子的发育——着重阐明质体在生殖细胞和营养细胞中的分布和变化

玉竹(Polygonatum simizui Kitag)小孢子在分裂前,质体极性分布导致分裂后形成的生殖细胞不含质体,而营养细胞包含了小孢子中全部的质体。生殖细胞发育至成熟花粉时期,及在花粉管中分裂形成的两个精细胞中始终不含质体。虽然生殖细胞和精细胞中都存在线粒体,但细胞质中无DNA类核。玉竹雄性质体的遗传为单亲母本型。在雄配子体发育过程中,营养细胞中的质体发生明显的变化。在早期的营养细胞质中,造粉质体增殖和活跃地合成淀粉。后期,脂体增加而造粉质体消失。接近成熟时花粉富含油滴。对百合科的不同属植物质体被排除的机理及花粉中贮藏的淀粉与脂体的转变进行了讨论。     

甘薯细胞质传递的研究：精细胞的质体和线粒体及其DNA存在的状况

甘薯（ＩｐｏｍｏｅａｂａｔａｔａｓＬａｍ．）成熟花粉为二细胞型,在授粉后萌发之前生殖细胞分裂形成精细胞。仍在花粉粒中的两个精细胞大小和形状基本相似,细胞质中含丰富的质体和线粒体。细胞质ＤＮＡ特异荧光显示精细胞及产生它们的前细胞———生殖细胞中均含有丰富的类核。一对精细胞中类核的数量无明显的差异。精细胞中存在两种形态类核,大而荧光强的类核可能为质体类核,而小的荧光弱的类核为线粒体类核。双亲或父系质体遗传在被子植物中是少数,本研究结果为旋花科的除牵牛属和打碗花属外又提供了新的一属具这种遗传方式的细胞学证据。     

Cytological Basis of Plastid Inheritance in Phaseolus vulgaris-Investigations of Plastids,Mitochondria and Their DNA Nucleoids During Pollen Development

Electron microscopic and DNA fluorescence microscopic observations of the plastids, mitochondria and their DNA in the developing pollen of Phaseolus vulgaris L. have demonstrated that the male plastids were excluded during microspore mitosis. The formed generative cell was free of plastids because of regional localization of plastids in early developing microspore and the extremely unequal distribution during division. The fluorescence observations of DNA showed that cytoplasmic (plastid and mitochondria) nucleoids degenerated and disappeared during the development of microspore/pollen, and were never presented in the generative cell at different development stages. These results provided precise cytological evidence of maternal plastid inheritance in Phaseolus vulgaris, which was not in accord with the biparental plastid inheritance identified from early genetic analysis. Based on authors' previous observations in a variety of common bean that the organelle DNA of male gamete was completely degenerated, the early genetic finding of the biparental plastid inheritance was unlikely to be effected by genotypic difference. Thus those biparental plastid inheritance might be caused by occational male plastid transmission, and plastid uniparental maternal inheritance was the species character of Phaseolus vulgaris.     

CYTOLOGICAL BASIS FOR CYTOPLASMIC INHERITANCE IN PSEUDOTSUGA MENZIESII. I. POLLEN TUBE AND ARCHEGONIAL DEVELOPMENT

Douglas fir (Pseudotsuga menziesii (Mirb.) Franco) ovules were used to study the method of pollen tube formation and penetration of the nucellus, the movement of the body cell down the pollen tube and development of the archegonia. No pollination drop forms but nucellar tip cells produce a minute secretion that may initiate pollen tube formation. Pollen tubes penetrate the nucellus causing degeneration of nucellar cells in contact with the pollen tube tip. The body cell becomes highly lobed and the tube cytoplasm forms thin sheets between the lobes. This may be the mechanism by which the large body cell is pulled down the narrow pollen tube. Body cell plastids and mitochondria remain unaltered during pollen tube growth, whereas tube cell organelles show signs of degeneration. The pollen tube penetrates the megaspore wall and settles in the archegonial chamber. During pollen elongation and pollen tube growth the egg matured. Egg cell plastids were transformed into large inclusions which filled the periphery of the egg while mitochondria migrated to the perinuclear zone. The neck cells, ventral canal cell and archegonial jacket cells are described. The significance of the body cell and egg cell ultrastructure is discussed in light of recent restriction fragment length polymorphism studies of plastid and mitochondrial inheritance in the Pinaceae.     

牵牛花受精前后卵细胞质体和线粒体及其细胞质DNA的变化

应用透射电镜研究了圆叶牵牛（Ｐｈａｒｂｉｔｉｓｐｕｒｐｕｒｅａ （Ｌ．） Ｖｏｙｇｈｔ）和大花牵牛（Ｐ． ｌｉｍ ｂａｔａ Ｌｉｎｄｌ．）卵细胞在受精前和受精后质体和线粒体的变化。观察到卵细胞在受精前高度液泡化,在核周及卵细胞周围的薄层细胞质中有质体和线粒体。质体含１～２ 个较大淀粉粒。线粒体多呈环形或杯状。卵细胞受精后,细胞质增多,质体的数量也明显增加。质体基质变得更加浓厚,并且普遍含有嗜锇的球体。合子中线粒体丰富,但缺乏卵细胞中那种环形或杯状线粒体,多呈圆形。细胞质ＤＮＡ检测的结果表明,卵细胞质ＤＮＡ荧光有大小和形状不同的两类,它们在细胞中随机分布。一类较大,呈环状,可能是线粒体中的ＤＮＡ显示的荧光;另一类小的点状荧光,可能大多是质体ＤＮＡ荧光,前者比后者多。卵细胞受精后,细胞质类核发生变化,表现在数量上明显地比受精前少。研究揭示了牵牛花合子中细胞质ＤＮＡ 减少的现象,说明了母系质体ＤＮＡ 在受精后可能被降解,提供了母系质体不传递到后代的可能机制。     

黑麦雄性细胞发育过程中细胞器DNA动态的荧光研究

DAPI(4’,6-diamidino-2-phenylindole)是一种DNA特异结合的荧光染料,可以用于在荧光显微镜下观察和检测各种DNA,尤其是细胞内含量甚微的DNA,包括质体DNA和线粒体DNA,其灵敏性和可靠性是被公认的,并得到了越来越多的Southern杂交实验的证明,而且实验操作简便易行。近几年,DAPI荧光技术已在细胞质遗传的研究领域获得了成功的应用。     

八倍体小偃麦与硬粒小麦及其杂种F_1的小孢子发生和花粉发育的细胞学研究

用压片法对八倍体小偃麦与硬粒小麦及其杂种F_1小孢子发生和花粉发育进行了详细的细胞学观察。结果表明,两亲本八倍体小偃麦和硬粒小麦的小孢子发生和花粉发育是基本正常的,二者结实率也基本正常。[八倍体小偃麦×硬粒小麦]F_1小孢子发生过程非常紊乱,花粉母细胞减数分裂中期Ⅰ出现较高频率的单价体和多价体,其相对紊乱系数高达0.67;减数分裂晚后期,每个细胞落后染色体平均3.56条;四分体期,每个四分体微核数平均2.82个。在杂种F_1花粉发育的每个时期都可产生不同频率的败育花粉。在三细胞成熟花粉期,杂种F_1可育花粉百分率为67.36%,基本能够满足传粉受精的需要,但其结实率仅为2.79%,远远低于两亲本结实率。因此杂种F_1花粉的败育不是导致其结实率低的主导因素。     

CYTOLOGICAL BASIS FOR CYTOPLASMIC INHERITANCE IN PSEUDOTSUGA MENZIESII. II. FERTILIZATION AND PROEMBRYO DEVELOPMENT

Douglas fir (Pseudotsuga menziesii [Mirb.] Franco) ovules were used to study male gamete formation, insemination of the egg, and free nuclear and cellular proembryo development. Two male nuclei form as the pollen tube either reaches the megaspore wall or as it enters the archegonial chamber. No cell wall separates them. They are contained within the body-cell cytoplasm. A narrow extension of the pollen tube separates the neck cells and penetrates the ventral canal cell. The pollen tube then releases its contents into the egg cytoplasm. The two male gametes and a cluster of paternal organelles (plastids and mitochondria) migrate within the remains of the body-cell cytoplasm toward the egg nucleus. Microtubules are associated with this complex. The leading male gamete fuses with the egg nucleus. The zygote nucleus undergoes free nuclear division, but the cluster of paternal organelles remains discrete. Free nuclei, paternal and maternal nucleoplasm, maternal perinuclear cytoplasm, and the cluster of paternal organelles migrate en masse to the chalazal end of the archegonium. There, paternal and maternal organelles intermingle to form the neocytoplasm, the nuclei divide, and a 12-cell proembryo is formed. The importance of male nuclei or cells, the perinuclear zone, and large inclusions in cytoplasmic inheritance are discussed in the Pinaceae and in other conifer families. This completes a two-part study to determine the fate of paternal and maternal plastids and mitochondria during gamete formation, fertilization, and proembryo development in Douglas fir.