大叶杨配囊及胚珠的形成和发育

本文应用细胞化学方法研究了大叶杨胚珠、胚囊的形成和发育过程中核酸、蛋白质及不溶性多糖的分布和消长。大孢子母细胞、大孢子四分体及功能大孢子中含较少不溶性多糖,但却含丰富的RNA和蛋白质。功能大孢子经分裂发育成八核的蓼型胚囊。四核胚囊开始积累细胞质多糖,成熟胚囊中除反足细胞外充满淀粉粒。反足细胞形成后不久即退化。助细胞具多糖性质的丝状器,受精前两个助细胞退化。卵细胞核对Feulgen反应呈负反应。二极核受精前由胚囊中部移向卵器,与卵器接触后融合形成次生核。发育早期的胚珠为厚珠心,双珠被。晚期,内珠被退化,故成熟胚珠为单珠被。四核胚囊时期,珠孔端珠心组织退化,胚囊伸向珠孔形成胚囊喙。合点端珠心组织含丰富的蛋白质和核酸,这一性质与绒毡层性质相似,可能涉及胚囊的营养运输。胚囊的营养来源于子房和胎座细胞内贮存的淀粉粒。     

天竺葵雌性生殖单位的超微结构

应用透射电镜研究了临近受精时天竺葵(Pelargonium hortorum Bailey)胚囊中的卵细胞、助细胞和中央细胞的结构。证明了卵细胞与助细胞以及助细胞与助细胞之间从合点端至珠孔端有很大的面积以质膜分界,仅珠孔端少部分以壁分隔。卵细胞与中央细胞之间同样缺乏细胞壁。在卵细胞的合点端,两质膜不同程度地分离形成宽窄相间的间隙。在间隙的絮状基质中存在小泡,这些小泡的产生似与卵和中央细胞中周质内质网的活动有关。推测小泡为多糖性质,可能为合子新壁的建造提供物质。卵细胞质中含巨大线粒体,质体和内质网也较丰富。基于超微结构的特征,可认为卵细胞具高度的生理合成活动的潜能。中央细胞极核位于珠孔端与卵器细胞毗邻,有利于在双受精作用中同时发生精细胞与卵细胞和精细胞与中央细胞核的融合。中央细胞的侧壁在珠孔端形成内突,具传递细胞的特点,表明这是雌配子体向孢子体摄取营养的重要部位。助细胞的细胞质含丰富的细胞器,这与多数植物中的相似,但具几个明显的特征,即核中存在微核仁,内质网形成圆球体或脂体,线粒体富集在丝状器的附近。传粉后花粉管进入胚囊之前,两个助细胞中一个退化。     

水稻胚囊超微结构的研究

水稻(Oryza sativa L.)胚囊成熟时,卵细胞的合点端无细胞壁,核居细胞中部,细胞器集中在核周围,液泡分散于细胞周边区域。助细胞珠孔端有丝状器,合点端无壁,核位于细胞中部贴壁处,细胞器主要分布在珠孔端,液泡主要分布在合点端。开花前不久,一个助细胞退化。中央细胞为大液泡所占,两个极核靠近卵器而部分融合,细胞器集中在极核周围和靠近卵器处,与珠心相接的胚囊壁上有发达的内突。反足细胞多个形成群体,其增殖主要依靠无丝分裂与壁的自由生长,反足细胞含丰富活跃的细胞器,与珠心相接的壁上有发达的内突。开花后6小时双受精已完成,合子和两个助细胞合点端均形成完整壁。合子中开始形成多聚核糖体、液泡减小。退化助细胞含花粉管释放的物质,其合点端迴抱合子。极核已分裂成数个胚乳游离核,中央细胞中细胞器呈活化状态。反足细胞仍在继续增殖。讨论了卵细胞的极性、助细胞的退化、卵器与中央细胞间界壁的变化、反足细胞的分裂特点等问题。     

番茄受精作用及其间隔期的研究

利用常规石蜡切片法研究了番茄受精作用的全过程,具体研究结果为:(1)授粉后2 h,花粉粒在柱头上萌发;约2~4 h,花粉管长入柱头,且末端膨大;约8 h后,生殖细胞进入分裂期;并于约两小时后,分裂为两个精细胞。(2)约14 h,花粉管进入子房腔;约18~24 h,花粉管进入胚囊,破坏一个助细胞,并在其珠孔端释放两个精子;随后被释放的精子移到卵细胞与次生核附近。(3)授粉后约30 h精核进入卵细胞;约34 h,精核与卵核融合,并在卵核内出现分散的雄性染色质,进而出现雄性核仁;44~50 h,雌、雄性核仁融合,形成合子;合子的休眠期为10 h左右。60 h之后,合子分裂形成二细胞原胚。(4)约26 h,另一个精子的精核与次生核核膜相贴伏,随后与之融合;约30~34 h,次生核内出现分散的雄性染色质,随之出现雄性核仁;约38~42 h,雌、雄性核仁融合,形成初生胚乳核。约44 h后,初生胚乳核进行有丝分裂,形成两个胚乳细胞。番茄胚乳发育属于细胞型。初生胚乳核无休眠期。(5)精子与次生核的融合比与卵核的融合快。(6)番茄的受精作用属于有丝分裂前配子融合类型。     

甜菜无融合生殖单体附加系M14成熟胚囊的超微结构特征

以甜菜无融合生殖单体附加系M14（Betavulgaris,2n=18＋1）为实验材料,利用电子显微镜技术对成熟胚囊及其超微结构进行研究。结果表明：M14成熟胚囊包括1个卵细胞、2个退化的助细胞、1个具有次生核的中央细胞和3-6个反足细胞。其卵细胞具有3种不同的形态：（1）极性正常的卵细胞,细胞核位于合点端,细胞质含有大量核糖体、线粒体、内质网等细胞器;（2）细胞核位于细胞中央;（3）细胞核位于珠孔端,且后2种形态细胞器的种类与数量少。大多数胚囊中的2个助细胞在开花前已退化。中央细胞的次生核位于反足细胞附近;未经受精自发分裂前的卵细胞与中央细胞的细胞核大、核仁明显,细胞器的种类与数量多,呈现旺盛代谢活动特征,成为二倍体孢子无融合生殖过程中,卵细胞与次生核自发分裂的细胞学标志。     

羊草受精作用及其胚与胚乳早期发育的观察

利用常规石蜡制片方法研究了羊草受精过程及胚与胚乳的早期发育,其主要结果为：(1)授粉后1h,花粉管破坏1助细胞,释放2精子。精子为眼眉状,难以区分其细胞质鞘;(2)授粉后1～2h,2个精子分别移向卵细胞与极核;(3)授粉后2～3h,精核分别贴附于卵细胞与极核核膜上;(4)授粉后3～10h,精核与卵核融合,并出现雄性核仁,形成合子;(5)授粉后3～4h,精核与极核融合,并出现雄性核仁,形成初生胚乳核,精核与极核的融合比与卵核融合快;(6)传粉后20h,合子分裂,合子的休眠期为10h左右;(7)传粉4h,初生胚乳核分裂,初生胚乳核没有休眠期;(8)羊草双受精作用属于有丝分裂前配子融合类型;(9)胚胎发育属于紫菀型,胚乳发育属于核型胚乳。     

甜菜无融合生殖单体附加系M14 成熟胚囊的超微结构特征

以甜菜无融合生殖单体附加系M14(Beta vulgaris, 2n=18+1)为实验材料, 利用电子显微镜技术对成熟胚囊及其超微结构进行研究。结果表明: M14成熟胚囊包括1个卵细胞、2个退化的助细胞、1个具有次生核的中央细胞和3-6个反足细胞。其卵细胞具有3种不同的形态: (1)极性正常的卵细胞, 细胞核位于合点端, 细胞质含有大量核糖体、线粒体、内质网等细胞器; (2)细胞核位于细胞中央; (3)细胞核位于珠孔端, 且后2种形态细胞器的种类与数量少。大多数胚囊中的2个助细胞在开花前已退化。中央细胞的次生核位于反足细胞附近; 未经受精自发分裂前的卵细胞与中央细胞的细胞核大、核仁明显, 细胞器的种类与数量多, 呈现旺盛代谢活动特征, 成为二倍体孢子无融合生殖过程中, 卵细胞与次生核自发分裂的细胞学标志。     

软毛猕猴桃的早期胚胎发生的研究

本文报道软毛猕猴桃的早期胚胎发生的研究:——1.子房上位,具34—45个心皮,每心皮含11—45个胚珠。胚珠倒生,单珠被,薄珠心。孢原为1—3个细胞组成,仅有一个转变为大孢子母细胞,其余的退化。2.成熟花粉粒为二细胞型。胚囊蓼型。在大多数胚囊中,两个极核受精前合并为次生核,少数到受精时仍不合并。一个助细胞为花粉管所破坏,另一个在受精期间解体,个别的保持到初生胚乳核分裂时期。3.根据观察,在北京地区,软毛猕猴桃的双受精大多发生在授粉后30—72小时内。受精时,一个精子同卵细胞核融合,另一个同次生核融合,其融合过程较卵核和精子融合结束的早。4.胚乳发育为细胞型。     

星星草受精作用及其胚与胚乳早期发育的观察

利用常规石蜡切片法对星星草[Puccinellia tenuiflora(Griseb．)Scribn．et Merr．]受精过程及胚与胚乳的早期发育进行了观察,主要结论如下：(1)开花后2h,花粉管破坏1个助细胞,释放2个精子,精子呈逗点状。(2)开花后2～3h,2个精子分别移向卵细胞与极核。(3)开花后3～5h,精核分别贴附于卵细胞与极核的核膜上。(4)开花后5～10h,精核与卵核融合,雄性核仁出现,合子形成。(5)开花后5～6h,精核与极核融合,并出现雄性核仁,形成初生胚乳核,精核与极核的融合比与卵核融合要快。(6)开花后20h左右,合子分裂。(7)开花后8h,初生胚乳核。     

韭菜胚囊发育与胚胎发生

韭菜胚囊发育为葱型,胚胎发生属柳叶菜型。成熟胚囊中,三个反足细胞形态上常类似卵器,其中二个呈助细胞状,一个呈卵细胞状。卵状反足细胞可分裂成多细胞原胚,但随着胚乳的发育而退化。在未受精胚囊中,卵细胞和卵状反足细胞均可分裂,它们的发生过程与合子胚相似,但因无胚乳哺育,均不能继续发育。论证了反足细胞胚的性质,初步探讨了胚乳与反足细胞无配子生殖的关系。     

Fertilization and Histochemical Investigation on Pulsatilla chinensis (Bung) Regel

Fertilization and variation of protein and starch grains in Pulsatilla chinensis (Bung) Regel have been studied at light microscopic level with histochemical test. Based upon the observations, the main conclusions are summarized as follows: The mature pollen grains are two-celled in which the generative cell shows the stronger protein staining than the vegetative cell. And vegetative cells are full of starch garins. When the pollen tube enters into the embryo sac, one synergid is destroyed, or in a few cases synergids are intact. Occasionally two synergids are disorganized as pollen tube penetrates. However, most of the remaining syuergids break down during fertilization, only in a few cases it remains till early stage of embryo development. The contents discharged by the pollen tube consist of two sperms, which stain intensely blue with protein dyes, a great amount of protein and starch grains. Mature female gametophyte (embryo sac) consists of an egg apparatus, central cell, which has a huge secondary nucleus, and antipodal apparatus which retain in course of fertilization. A few of embryo sac contain two sets of egg apparatus, a central cell with two huge secondary nuclei and two sets of antipodal apparatus. In some nucleoli of the central cell the comb-like structure pattern may be detected clearly. There are 1–2 small nucleoli in some egg cells and central cells. All the cells in embryo sac show protein positive reaction. According to the different shades of the color in cells, its may be arranged in the following order: antipodal cells, synergids, central cell and egg cell. Only a few small starch grains are present near nuclei of central cell and egg cell before fertilization, but no starch grains remain in most of the central cell, the synergids and antipodal cells. The fertilization is of the premitotic type. The fusion of the sexual nuclei progresses in the following order: 1, sperms approach and lie on the egg nucleus and secondary nucleus; 2, sperm chromatin sinks themselves into female nucleus, and male nucleolus emerges with the sperm chromosome; and 3, male nucleoli fuse with the nucleoli of egg nucleus and central cell nucleus, and finally forming the zygote and the primary endosperm cells respectively. Nevertheless, as it is well known, the fertilization completes in central cell obviously earlier than that in egg cell. Though it has been explained in cereals and cotton, in Pulsatilla chinensis the main reason is that nucleolar fusion of the male and female nucleoli in egg nucleus is slower than that in secondary nucleus. And the dormancy of the primary endosperm nucleus is shorter than that of the zygote. In the process of fertilization, histochemical changes are considerably obvious in the following three parts: 1, from the begining of fusion of male and female nuclei to form zygote and primary endosperm cell, Protein staining around female nucleus appears to increase gradually; 2, no starch grains are detected in embryo sac. Though only starch grains are carried in by pollen tube, they are completely exhausted during this period; and 3, near completion of fertilization starch grains appear again in zygote, however, not yet in primary endosperm nucleus till its dividing for the first time. The present study reveals that antipodal cells and synergids seem to play a significant role in nutrition of the embryo sac during the fertilization.     

土麦冬离体萌发花粉管中生殖细胞与营养核的动态变化

主要报道了土麦冬人工培养萌发花粉管中生殖细胞与营养核的动态变化。多数花粉管中,生殖细胞与营养核贴合后,开始进行有丝分裂,贴合时,营养核略呈弥散状态。在分裂早中期,生殖细胞与营养核分开,从贴合到分开大约经历３－５ｈ,精子形成后,不与营养核连接。ＤＡＰＩ对生殖细胞的有丝分裂有抑制作用。少数花粉管中,生殖细胞核进行无丝分裂,有缢裂和劈裂两种方式。生殖细胞核发生缢裂的花粉管中,未观察到生殖细胞与营养核的贴     

The initiation and organization of microtubules in germinating pear (Pyrus communis L.) pollen

To study microtubule organization in germinating pear (Pyrus communis L., cv., Bartlett) pollen, we removed the pollen wall by freeze-fracturing before treating the resultant pollen protoplasts by conventional immunofluorescence procedures. Results reveal that axial bundles of microtubules are present in the generative cell of both inactivated and activated pollen grains. Microtubules are not present in the vegetative cells of inactivated pollen, but they are present in the vegetative cells of activated pollen grains. Microtubule nucleation occurs in the vegetative cell cortex. Subsequently, the microtubules grow as branching arrays through most of the vegetative cell cortex except at the apertures where they form localized converging or criss-cross patterns. Eventually, in a germinated pollen grain, the microtubules form network-like arrays through most of the pollen grain and a collar of short arrays at the base of the pollen tube. It is suggested that the role of vegetative cell microtubules in pollen germination is indirect through their mediation of the conformational changes in actin organization that are essential for pollen germination.     

An autoradiographic study of RNA synthesis during maturation and germination of pollen grains ofHyoscyamus niger

Summary The pattern of RNA synthesis during maturation and germination of pollen grains ofHyoscyamus niger was studied using³H-uridine autoradiography. Incorporation of label during pollen maturation was periodic with peak RNA synthesis occurring in the uninucleate, nonvacuolate pollen grains and in the vegetative cell of the bicellular pollen grains. During the early stages of germination, isotope incorporation occurred predominantly in the nucleus of the vegetative cell with little or no incorporation in the generative cell. With the appearance of the pollen tube, incorporation of³H-uridine in the vegetative cell nucleus decreased and completely disappeared at later stages of germination. No incorporation of isotope was observed in the sperms formed in the pollen tube by the division of the generative cell. From a comparison of the results of this study with those of previous works on RNA synthesis during pollen embryogenesis in cultured anthers ofH. niger, it is concluded that in contrast to embryogenic development, there is no requirement for sustained RNA synthesis by the generative cell nucleus for normal gametophytic development.     

Differential localization of the LIM domain protein PLIM-1 in microspores and mature pollen grains from sunflower

 PLIM-1 is a LIM domain protein specifically expressed in pollen grains. Using two PLIM-1-specific monoclonal antibodies we studied its expression and intracellular location at various developmental stages of sunflower (Helianthus annuus L.) pollen. Our studies show that the protein appears at the microspore stage in a limited number of cytoplasmic bodies, becomes undetectable in bicellular pollen, and reappears in tricellular pollen grains in cortical patches particularly concentrated in the F-actin-enriched germination cones of the vegetative cell. The developmental stage-dependent, different location of the protein suggests a dual function during pollen development. While this function in microspore development remains obscure, the high concentration of PLIM-1 in the germination cones of mature pollen suggests that it participates in the germination process as well as in pollen tube growth. Received: 11 August 1998 / Revision accepted: 15 December 1998     

Ultrastructure of the sperms ofPlumbago zeylanica

Summary Male gametes ofPlumbago zeylanica were examined in pollen grains and tubes using light and electron microscopy of chemically and physically fixed tissues, and Nomarski interference microscopy of isolated, living sperm cells. Male gametes are elongate, spindleshaped cells containing a nucleus, mitochondria, ER, ribosomes, vesicles, dictyosomes, probable microfilaments, and a variable number of plastids. In mature pollen grains ofP. zeylanica, the two sperm cells are directly linked; they share a transverse cell wall with plasmodesmata and are enclosed together by the inner vegetative cell plasma membrane. One of these two sperms is also associated with the vegetative nucleus as a consistent feature of pollen grain organization. The basis of this association appears to be a long, narrow projection of the sperm cell (averaging < 1 m wide and about 30 m long) which wraps around the periphery of the vegetative nucleus and occupies embayments of that nucleus. This association is maintained throughout pollen tube growth but becomes less extensive near the completion of tube growth and is severed following tube discharge. The consistent occurrence of the sperm-vegetative nucleus association in pollen grains, tubes and isolated pollen cytoplasm suggests that the two structures may be directly connected, but attempts to visualize this type of connection were unsuccessful. Possibly, the entwining nature and extent of complementary interfaces between vegetative nucleus and sperm may have a role in stabilizing their association. Functionally, the two sperms and vegetative nucleus appear to travel as a linked unit within the pollen tube, possibly increasing the effectiveness of gamete delivery and helping to ensure nearly simultaneous transmission of sperms into the receptive megagametophyte.     

Studies on Fertilization and Embryogenesis of Red Vinespinach (Basella rubra L.)

This paper presents detailed report on the process of fertilization and the develop- ment of embryo and endosperm of Basella rubra L. The results obtained are summarized as follows: About 4–6 hours after anthesis a great deal pollen grains germinated on the stig- ma; 6–11 hours, the pollen tube passed through the style; 11–16 hours, the pollen tube reached the ovary cavity. About 16–18 hours, one sperm nucleus entered into the egg and the other one entered into the secondary nucleus. In most cases, after 16–24 hours the double fertilization had completed. After 2–8 days, two-celled proembryo was first shown. Finally, the proembryo gave rise to multicellular globular embryo proper. The development of the embryo of Basella rubra L. conforms to the Asterad type. Whether the Asterad type had a high frequency needs further to be studied. Although the fertilization of the sperm nucleus with the secondary nucleus began later, the fusion of two sexual nuclei and the development of the endosperm proceeded often quickly. After 20 hours, the free nuclei period began, and after 2–4 days the free nuclei of endosperm were rapidly formed. The endosperm of Basella rubra L. is a nuclear type. And at the time When the seed had ripened the endosperm tissues were all absorbed by the developing embryo. The author observed that there are different distributions of the vegetative nucleus and two sperms in the pollen tube, and that two sperms entered a egg or secondary nucleus. The changes of the starch accumulated and distributed et al. in the floral organs are also studied and discussed.     

The pollinium ofLoroglossum hircinum (Orchidaceae) between pollination and pollen tube emission

The structure of the massulae composing the pollinium ofLoroglossum hircinum was studied before pollination and 12 and 24 hours afterwards. The grains are grouped in tetrads closely packed in massulae. The exine is only present on the outside of the massulae. The intine consists of two layers: a compact layer surrounding the pollen grain and a looser layer surrounding the pollen grain and a looser layer surrounding the tetrad. Twelve hours after pollination, pollen volume and the space between the tetrads increase due to vacuolization. Twenty-four hours after pollination, pollen volume and tetrad spacing are higher due to vacuolization and some grains have emitted pollen tubes. Pollen growth due to vacuole formation, and the absence of common walls between adjacent tetrads lead to crumbling of the massulae. The mature pollen grain does not have apertures: the site of pollen tube emission is determined after pollination. The first grains to germinate are those in the centre of the massula. The vegetative cell nucleus is the first to enter the pollen tube; the generative cell elongates and undergoes the second haploid mitosis shortly after entering the pollen tube.