杜鹃雌雄配子体的超微结构及受精过程中精细胞质的传递

杜鹃雌雄配子体的超微结构及受精过程中精细胞质的传递国凤利,胡适宜（北京大学生命科学学院、北京１００８７１）应用透射电镜及ＤＮＡ特异性荧光技术对杜鹃的雌雄配子体进行了研究。在比较了受精前两性配子中细胞器特征的基础上,进一步鉴定了受精以后合子及胚乳细胞中...     

棉花751与多父本杂交受精细胞学研究及性状分析

本文报导了以棉花７５１为母本与鸡脚叶棉、乌干达棉、红叶棉、汉寿大桃、草棉等５个父本的混合花粉进行杂交。授粉后,观察了７５１大小孢子和雌雄配子体的发生、发育及其受精过程。切片观察表明在大小孢子的发生、发育与花蕾大小成正相关。大小孢子的发生在减数分裂前是不同步的。小孢子的发生早于大孢子发生１￣２个时期。减数分裂后它们的发育逐渐趋向同步。开花时达到同步成熟。用多父本的混合花粉授粉,其受精过程仍为单精入卵     

被子植物的胚胎发育

被子植物是植物界中的高级优势类群,在世代交替中,孢子体占绝对优势,具有高度的器官分化和广泛的适应性。雌雄配子体都简化到只有几个细胞,寄生在孢子体上,得到严密的保护和充分的营养。在有性繁殖中有双受精现象,胚和胚乳都是受精后的产物。卵核受精后的合子发育成胚,受精后的极核发育成胚乳;胚珠形成种子;膨大的子房发育成果实。种子萌发后,幼小的胚逐渐发育为成长的孢子体。所以,合子是孢子体的第一个细胞。     

高等植物花器官的特异性基因

生殖过程是高等植物生活史中最重要的阶段,它不能仅被看作是一个精子与卵细胞结合的简单过程。实际上它包括雌雄配子体的发育、雄配子体与雌配子体的识别及相互作用、精子与卵细胞的识别及相互作用等一系列复杂的生理生化变化。对于被子植物来说,还存在着双受精这样一个更为复杂的过程。因此,作为这一生理过程的主要执行器官,花器官的生长发育就格外受到生物学家们的重视。早期对于花器官的研究主要集中     

花生雌雄两性细胞发育关系的研究

本试验结果表明,雌雄配子体形成过程和程序与前人结论相一致,但发现：1)尽管雌雄蕊同时形成,但在雌雄配子形成不同步的情况下,花粉和胚囊中的大量淀粉却能同时充实,并且同时成熟等待受精。2)大孢子母细胞形成比小孢子母细胞迟0.7～0.8片展叶;雌配子体形成比雄配子体迟0.5～0.6片展叶。3)雌雄两性细胞发育特点不同,雄性是小孢子形成快而花粉粒发育慢,雌性是大孢子形成慢而胚囊发育快,为此在所用时间上,花粉粒发育约是小孢子形成的1.6倍,是胚囊发育的2.3倍,而大孢子形成约是胚囊发育的2.2倍,约是小孢子形成的1.4倍。     

款冬的胚胎学

系统报道了中藏药款冬大小孢子的发生、雌雄配子体发育和胚胎的发育过程,药壁发育双子中型,绒毡层发育接近菊科中的“The Comos bipinnatus”型;成熟花粉为3细胞型;单株被,薄珠心,倒生胚珠,具发达的珠被绒毡层;胚囊发育4孢子型,接近五福花（Adoxa)型胚囊,受精作用属于有丝分裂前配子体融合类型,胚乳发育为核型,胚胎发育紫菀型千里光变型。讨论了该种胚胎学特征的生态学及系统学意义。     

蕨类植物孢子与种子植物花粉萌发的比较

蕨类植物孢子与种子植物花粉在有性生殖过程中都具有重要的作用。花粉作为种子植物的雄配子体, 通过萌发后极性生长的花粉管将精细胞送到胚囊完成受精作用。蕨类植物孢子作为配子体的原始细胞, 通过不对称的有丝分裂产生一大一小两个细胞, 小细胞萌发出极性生长的假根, 大细胞继续分裂发育为原叶体(配子体)。成熟的花粉和蕨类植物孢子都是代谢高度静止的细胞, 两者的萌发过程不仅都受到各种不同环境因子的影响, 而且在信号转导、极性建立和能量代谢等方面可能有着相似的调控机制。本文综述了蕨类植物孢子和种子植物花粉萌发过程的差异和保守性特征。     

蕨类植物孢子与种子植物花粉萌发的比较

蕨类植物孢子与种子植物花粉在有性生殖过程中都具有重要的作用。花粉作为种子植物的雄配子体,通过萌发后极性生长的花粉管将精细胞送到胚囊完成受精作用。蕨类植物孢子作为配子体的原始细胞,通过不对称的有丝分裂产生一大一小两个细胞,小细胞萌发出极性生长的假根,大细胞继续分裂发育为原叶体（配子体）。成熟的花粉和蕨类植物孢子都是代谢高度静止的细胞,两者的萌发过程不仅都受到各种不同环境因子的影响,而且在信号转导、极性建立和能量代谢等方面可能有着相似的调控机制。本文综述了蕨类植物孢子和种子植物花粉萌发过程的差异和保守性特征。     

樱桃品种‘拉宾斯’雌雄配子体发育的解剖学研究

以‘拉宾斯’、‘艳阳’、‘马哈利CDR-1’为材料,对其自花授粉坐果率进行统计分析。用石蜡切片技术对拉宾斯雌雄配子体发育过程进行解剖观察。结果表明：(1)‘拉宾斯’自花授粉坐果率明显低于‘艳阳’和‘马哈利CDR-1’。(2) 拉宾斯从小孢子母细胞形成至成熟花粉的各发育阶段均观察到小孢子退化、花粉囊中的花粉干瘪、出现空腔。花粉成熟阶段部分花药绒毡层细胞不发生退化,花粉难以散出,造成雄性败育。(3)雌配子体在大孢子母细胞发育阶段出现大孢子发育不良或不能形成等异常发育雌配子体败育类型。(4)雄配子体在3月1日左右开始发育,3月14日左右趋近成熟;雌配子体则发育相对较晚,3月12日左右开始发育,3月25日左右趋近成熟。研究表明,‘拉宾斯’雌、雄配子体在其形成发育过程中的各种异常使其不能正常受精并且雌雄配子体发育不同步,最终导致坐果量低下。     

春砂仁传粉及其生殖生物学研究进展(综述)

春砂仁是我国南方一种重要的中草药,受自身遗传特性和环境因素的影响,其自然结实率极低,严重制约了其医疗用药。为了探讨影响春砂仁产量的生物学机理,进而为生产上提高春砂仁果实产量提供理论基础,本文对春砂仁雌、雄配子体发育及传粉、受精过程、果实和种子发育等方面的研究状况进行概述,并对存在的问题和未来研究趋势进行展望。     

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.     

Fertilization in Pinus Bungeana

Pinus bungeana is a species endemic to China and as yet its embryology has not been reported. The present paper deals with its process of fertilization in some details. 1. The development of the male gamete and the structure of the archegonium. The spermatogenous cell has already divided into two uniqual male gametes in the middle of May (in 1978, at Peking), about ten days before fertilization. Both sperms are spheroidal to ellipsoidal. The larger sperm is about 94 × 65 μm and the smaller one, about 72 × 58 μm in size. As the pollen tube approaches the archegonium the two sperms move toward the apex of the tube together with the remaining contents. Generally the larger sperm precedes the smaller one. The cytoplasmic contents also contain a sterile cell, 3—43×2—29 μm in size and a tube nuleus, 15—30 μm in diamter, besides the sperms. A mass of starch grains of more or less similar to sperm in size is also included in the contents of the pollen tube. Generally 3—4, even up to 7–8 pollen grains germinate normally within an ovule. Therefore, many sperms (up to 14—16) may be present on the same nucellus. The archegonium is elongato-ellipsoidal, about 870 ×500 μm in size. Arehegonia are single, 2—(3—5) in number, with 2 neck cells and a layer of jacket cells. The central cell divided in the middle of May and gave rise to the ventral canal cell and the egg. As the archegonium matures the cytoplasm becomes radiate fibrillae around the egg nucleus. The egg nucleus is large, 150—226 μm in diameter. One large nucleolus, 22—25 μm in diameter and sometimes up to 50; small nueleoli are present within the nucleus. 2. Fertilization Pollination takes place in the first week of May and fertilization will be effected from the end of May to the first week of June of next year. The interval between pollinatin and fertilization in P. bungeana is about thirteen months and the lapse of time is almost similar to most of the Pinus so far recorded. When the pollen tube contacts the archegonium through the neck cells all its contents are discharged into the egg cell. Usually the larger sperm fuses with the egg nucleus and the rest of the contents stays in the upper part of the egg cell. It is interesting to note that the nonfunctional second sperm also moves toward the egg nucleus and often divides by mitosis; and this phenomenon is not reported elsewhere. At the earlier stage of the fusion between male and female nuclei the male nucleoplasm is dense and finely granular while the female nucleoplasm is thin and coarsely granular, hence the boundary between them is very clear. The nuclear membranes of both nuclei persist for a long time. After the male nucleus sinks into the female nucleus completely, both nuclei begin to divide and enter into the prophase and then the metaphase simultaneously. By this time the paternal and maternal chromosome sets with their spindles still remain at certain distance from each other. Then the paternal chromosomes with their spindle move gradually toward the maternal ones. At first a multipolar common spindle appears as the maternal and paternal spindles with their chromosomes merge together. Finally a regular bipolar spindle is formed and both the maternal and paternal chromosomes become arranged on the equatorial plate. In the meantime, the process of fusion is complete and the zygote is at the stage of metaphase. At the moment the spindle looks greater in width than in length, being about 80×65—70 μm in size. 3. Supernumerary nuclei and sperms. The ventral canal cell degenerates soon after its formation. While the supernumerary sperms divide usually after their entrance into the egg cell. Therefore, the supernumerary nuclei probably derive directly from the smaller sperms or indirectly from mitoses of the larger ones Generally the nucleoplasm of the supernumerary nuclei is rather thin while the nucleoplasm of the undivided sperms is rather dense. This shows that the former is in the state of degeneration. The supernumerary nuclei of P. bungeana are as many as 7, their usual size being 43—58×32—43 μm. In the upper part of some egg cells there are still secondary smaller sperms about the size of 36 × 29 μm, Their volume is just about half of the usual smaller sperm. Probably they are derived from the division of the smaller sperms.     

The Gametophytes and Fertilization in Pseudotaxus

The development of the gametophytes and fertilization of Pseudotaxus chienii Cheng has been investigated. Pollination first occurred on April 17 (1964). The pollen grains shed at the uninucleate stage and germination on the nucellus is almost immediate. The pollen tubes approached the freenucleate female gametophyte about May 5. The spermatogenous cell is continuously enlarging with the growth of the pollen tube and two unequal sperms are formed after its division. Occasionally the small sperm may divide further into two smaller ones. During pollination the megaspore mother cell is in meiosis and 3 or 4 megaspores are formed. Generally 2 or 3 megaspores at the micropylar end are going to degenerate while the chalaza] megaspore is rapidly enlarging. After 8 successive simultaneous divisions of the functional megaspore 256 free nuclei are resulted and they are evenly distributed at the bulge of the famale gametophyte. Then the wall formation follows. Sometimes there are more than two, even as many as 5–6 gametophytes developed within a single ovule. The archegonial initials become differentiated at the apical end of the female gametophyte. They are usually single and apical, rarely lateral in position. The number of the archegonia vary from 3 to 7, usually 4–6. There are 2–8 neck cells in each archegonium which is surrounded by a layer of jacket cells. The central cell divided about May 20–26 (1964) and the division of the central cell gives rise to the egg and the ventral canal nucleus, the latter being degenerated soon. There are many proteid vacuoles near the nucleus of the matured egg. The fertilization took place about May 23–26 (1964). At first, the pollen tube discharges its contents into the egg, then the larger sperm fuses with the egg nucleus in the middle part of the archegonium. At the same time the male cytoplasm also fuses with the female cytoplasm and a layer of densely-staining neocytoplasm is formed around the fused nucleus. The smaller sperm, tube nucleus and sterile cell usually remain in the cytoplasm above the egg nucleus for some time. Based upon the observations of the development of the gametophytes and fertilization the authors conclude that Pseudotaxus is more close related to Taxus than any other member of Taxaceae.     

Studies in Fertilization of Fokienia

Material of Fokienia hodginsii was collected in 1964 from Fengyangshan (alt. 1000–1400 M) in Lungchuan county, Chekiang province. This paper deals with the fertilization in Fokienia. It includs the structure of male and female gametes as wed1 as the process of fusion of their nuclei and cytoplasm respectively. The division of the spermatogenous cell of Fokienia occurred by the end of June (1964) and two sperms similar in shape and size were formed when pollen tube reached the top of archegonia. Two equalsperms look like two hemispherical bodies conjoined togather. The sperm possesses cell wall and is about 65 μ in diameter. Its nucleus is rather large and about 45–50 μ in diameter. There is a nucleolus in the nucleus. Outside the nucleus the dense cytoplasm forms the deep colored zone, some 10 μ in thickness. This zone is separated from the nucleus by a narrow perinuclear zone, and from the plasmalemma by a marginal zone. The perinuclear zone is about 2 μ thick, and the mariginal zone is from 3 to 4 μ thick. Both zones have transparent cytoplasm. When the archegonium is formed, the central cell has a small nucleus which is located below the neck ceils. At the middle of June (1964), the central cell divides to form the ventral nucleus and the egg nucleus. The egg nucleus sites primarily at the upper part of archegoninm and has only one nucleolus. Then the egg nucleus increases gradually in sim and moves to the central part of the archegoninm. In mature archegonium there are usually 4–5, rarely 6–7 nucleoli in the egg nucleus, each of them is about 15 μ in diameter. The egg cell in Fokienia hodginsii is about 500 in length. The female nucleus is larger than the male one. After egg cell matures, its cytoplasm increases gradually, while the central vacuole decreases gradually and almost disappears completely after fertilization. It is interesting to note that there are 1–2 dense cytoplasm masses at the upper or lower part of egg nucleus. The shape of the mass is similar to that of the egg nucleus but no membrane is formed. These cytoplasm masses are about 50–70 μ in diameter in some cases. The fertilization of Fokienia took place at the end of June when the growing tip of pollen tube had reached the top of the archegoninm. Then the neck cells become disorganized and degenerated. It is possible that all the cytoplasmic contents of pollen tubes are released into the archegoninm. Before fertilization, the cytoplasm around the sperms and sterile cell and tube nucleus are in front of these two sperms. Then the sperms separate from each other and come down into the cytoplasm of the egg. When the mede nucleus contacts with the egg nucleus, both become flattened along their contact surface. Then the nuclear membranes of both sperm and egg nuclei become ultimately disintegrated. Thus the fusion process is complete. However, it is nia, though the opposite is the case in an exceptional example. When the sperm nucleus passes into the cytoplasm of egg cell, its cytopasm is released inside the archegonium along with it. During the course of fusion of the male and female nuclei, tile fertilized nucleus is surrounded by both female and male cytoplasm. Thus the male cytoplasm along with the peripheral cytoplasm of the egg cell invests the two nuclei lying in contact and forms a dense neocytoplasm. When the zygote divides, the neoeytoplasm is full of the starch grains and a dense cytoplasm sheath is formed. After fertilization, the fused nucleus moves toward the base of the egg cell. It seems that the movement of the fused nucleus is not a simple mechanical movement but turned over repeatedly toward the base of the arehegonium. Sometimes the position of the sperm and egg nuclei makes a turn of 180. At the same time the track of the fertilized egg nucleus with vacuoles in the archegonium may be traced. After zygote moves into basal part of the archegonium, first intranuclear mitosis occurs. The nuclear envelop of zygote disappears gradually at the telophase of the first mitosis. Then division of the free nuclei of proembryo follows. From fertilization to the stage of proembryo formation, the second sperm may sometimes enter into the cytoplasm of the egg cell. Mitosis of the second sperm nucleus may take place in the upper part of the archegonium. In addition, there are often several supernmnerary nuclei (as many as 7–8 in number) in the same egg cell. These nuclei are also surrounded by dense cytoplasm. They may persist for some time and be recognizable at somewhat later stages of the proembryo or even after the elongated suspensors are formed. In some cases, there are some cell groups above the upper tier of proembryo. These cell groups are also surrounded by dense cytoplasm. Either the supernumerary nuclei or cells are surrounded by the dense cytoplasm. Probably they are derived from the mitosis or amitosis of the second sperm. Investigations on submicroscopic structures of sperm and egg in relation to the fertilization of Cupressaceae have been carried out extensively during the last decade. The fate of male cytoplasm has been debated for a long time and this problem attracted attention again in the nineteen seventies. At last the concept of neocytoplasm has been established soundly based upon the information from observation of electron microphotographs. The neocytoplasm is also visible under the light microscope though the components are not recognizable. The sperms of Fokienia are similar to those of Cupressus funebris, Juniperus communis, Sabina virginiana, Tetraclinis articulata, Chamaecyparis pisifera as well as the genus Thujopsis and others. Two sperms are all effective in fertilization and this is the common phenomenon of the family Gupressaceae.     

Development of Gametophytes in Fokienia Cupressaceae

The structure of the ovule and the development of the gametophytes in Fokienia, an endemic genus of Cupressaceae are described in some detail. Two wings, one small and one large, are developed along the micropylar end of the ovule and two resin canals are present in each of them. The material collected in the middle of April was already pollinated and the pollen began to germinate on the nucellus. The sterile cell, tube cell and spermatogenous cell have been formed in the tube in the first collection of April 17. At the end of June the division of the spermatogenous cell results in two sperms of Similar size and shape and the division plane is usually parallel to long axis of the pollen tube. Both sperms are effective in fertilization. 4096 free nuclei (actual counting, 3733—4224 ones) are produced through 12 times of repeated divisions of the functional megaspore, then cell walls appear among the free nuclei and cellular female gametophyte is formed. The number of archegonia varies from 6 to 16, mostly 9–12. The archegonial complex is enclosed by 2–3 layers of jacket cells. The neck cells are usually 4 in number, arranged in 1–2 layers. The central cell divides and results in the formation of one ventral canal nucleus and one egg nucleus. Fertilization takes place in the middle of the archegonium. The development of the gametophytes of Fokienia is more or less similar to that of Sabina.     

FERTILIZATION IN PLUMBAGO ZEYLANICA: GAMETIC FUSION AND FATE OF THE MALE CYTOPLASM

Developmental phases surrounding the processes of gametic delivery and fusion were examined ultrastructurally in the reduced megagametophyte of Plumbago zeylanica, which lacks synergids. Gametic delivery occurs at the end of pollen tube growth and results in deposition of two male gametes, a vegetative nucleus, and a limited amount of pollen cytoplasm between the egg and central cell. Discharge of these materials from the tube is accompanied by loss of inner and outer pollen tube plasma membranes, loss of sperm-associated cell wall components, and disruption of the formerly continuous cell wall between the egg and central cell. The dispersion of egg cell wall components directly exposes female reproductive cell membranes to the unfused male gametes and pollen tube without disrupting gametic cell plasma membranes. Presence of unfused sperms within the female gametophyte appears to be a transitory phenomenon, lasting less than 5 min at the end of over 8½ hr of pollen tube growth. At the time of gametic deposition, plasma membranes of unfused sperm cells become directly appressed to plasma membranes of both the egg and central cell. Gametic fusion is initiated by a single fusion event between membranes of participating male and female cells, which is rapidly followed by subsequent, secondary fusion events between the same two cells at different locations along their surface. Gametic fusion results in the transmission of male gamete nuclei with co-transmission of nearly the entire sperm cytoplasmic volume and organellar complement, and it is possible to identify heritable male cytoplasmic organelles within both the incipient zygote and endosperm. Paternally originating plastids may be distinguished from maternal plastids by differences in morphology and staining characteristics, whereas paternal mitochondria may be distinguished from maternal mitochondria by populational differences in mitochondrial size which are statistically significant. Such observations further indicate that transmitted paternal mitochondria seem to remain viable, as judged by their ultrastructural appearance, and are transmitted exclusively by sperm cytoplasm rather than discharged pollen cytoplasm. The presence of anucleate, membrane-bounded cytoplasmic bodies between the egg and central cell are identifiable on the basis of their enclosed organelles and indicate that fragmentation of a small amount of the sperm cytoplasm associated with the vegetative nucleus commonly occurs. The presence and identification of sperm cytoplasmic organelles and associated membranes within female reproductive cells following gametic transmission represents strong evidence in support of the cellular basis of nuclear and cytoplasmic transmission during sexual reproduction in Plumbago.     

水稻双受精过程的细胞形态学及时间进程的观察

应用常规石蜡切片和荧光显微镜观察水稻（Oryza sativa）受精过程中雌雄性细胞融合时的形态特征及时间进程,确定合子期,为花粉管通道转基因技术的实施提供理论依据。结果表明：授粉后,花粉随即萌发,花粉管进入羽毛状柱头分支结构的细胞间隙,继续生长于花柱至子房顶部的引导组织的细胞间隙中,而后进入子房,在子房壁与外珠被之间的缝隙中向珠孔方向生长,花粉与花粉管均具有明显的绿色荧光。花粉管经珠孔及珠心表皮细胞间隙进入一个助细胞,释放精子。精子释放前,两极核移向卵细胞的合点端：两精子释放于卵细胞与中央细胞的间隙后,先后脱去细胞质,然后分别移向卵核和极核,移向卵核的精核快于移向极核的精核：精核与两极核在向反足细胞团方向移动的过程中完成雌雄核融合。大量图片显示了雌雄性核融合的详细过程以及多精受精现象。水稻受精过程经历的时间表如下：授粉后,花粉在柱头萌发：花粉萌发至花粉管进入珠孔大约需要0．5小时：授粉后0．54,时左右,花粉管进入一个助细胞,释放精子：授粉后0．5—2．5小时,精卵融合形成合子：授粉后约10．0小时,合子第1次分裂,合子期为授粉后2．5-10．04,时：授粉后1．0-3．04,时,精核与两极核融合：授粉后约5．0小时,初生胚乳核分裂。’     

水稻双受精过程的细胞形态学及时间进程的观察

应用常规石蜡切片和荧光显微镜观察水稻(Oryz a sativa)受精过程中雌雄性细胞融合时的形态特征及时间进程, 确定合子期, 为花粉管通道转基因技术的实施提供理论依据。结果表明: 授粉后, 花粉随即萌发, 花粉管进入羽毛状柱头分支结构的细胞间隙, 继续生长于花柱至子房顶部的引导组织的细胞间隙中, 而后进入子房, 在子房壁与外珠被之间的缝隙中向珠孔方向生长, 花粉与花粉管均具有明显的绿色荧光。花粉管经珠孔及珠心表皮细胞间隙进入一个助细胞, 释放精子。精子释放前, 两极核移向卵细胞的合点端; 两精子释放于卵细胞与中央细胞的间隙后, 先后脱去细胞质, 然后分别移向卵核和极核, 移向卵核的精核快于移向极核的精核; 精核与两极核在向反足细胞团方向移动的过程中完成雌雄核融合。大量图片显示了雌雄性核融合的详细过程以及多精受精现象。水稻受精过程经历的时间表如下: 授粉后, 花粉在柱头萌发; 花粉萌发至花粉管进入珠孔大约需要0.5小时; 授粉后0.5小时左右, 花粉管进入一个助细胞, 释放精子; 授粉后0.5-2.5小时, 精卵融合形成合子; 授粉后约10.0小时, 合子第1次分裂, 合子期为授粉后2.5-10.0小时; 授粉后1.0-3.0小时, 精核与两极核融合; 授粉后约5.0小时, 初生胚乳核分裂。     

Paternal cytoplasmic transmission in Chinese pine (Pinus tabulaeformis)

Summary. In this paper, the stages of normal sexual reproduction between pollen tube penetration of the archegonium and early embryo formation in Pinus tabulaeformis are described, emphasizing the transmission of parental cytoplasm, especially the DNA-containing organelles – plastids and mitochondria. The pollen tube growing in the nucellus contained an irregular tube nucleus followed by a pair of sperm cells. The tube cytoplasm contained abundant organelles, including starch-containing plastids and mitochondria. The two sperm cells differed in their volume of cytoplasm. The leading sperm, with more cytoplasm, contained abundant plastids and mitochondria, while the trailing one, with a thin layer of cytoplasm, had very few organelles. The mature egg cell contained a great number of mitochondria, whereas it lacked normal plastids. At fertilization, the pollen tube penetrated into the egg cell at the micropylar end and released all of its contents, including the two sperms. One of the sperm nuclei fused with the egg nucleus, whereas the other one was retained by the receptive vacuole. Very few plastids and mitochondria of male origin were observed around the fusing sperm and egg nuclei, while the retained sperm nucleus was surrounded by a large amount of male cytoplasm. The discharged tube cytoplasm occupied a large micropylar area in the egg cell. In the free nuclear proembryo, organelles of maternal and paternal origins intermingled in the neocytoplasm around the free nuclei. Most of the mitochondria had the same features as those of the egg cell, but some appeared to be from sperm cells and tube cytoplasm. Plastids were obviously of male origin, with an appearance similar to those of the sperm or tube cells. After cellularization of the proembryo, maternal mitochondria became more abundant than the paternal ones and the plastids enlarged and began to accumulate starch. The results reveal the cytological mechanism for paternal inheritance of plastids and biparental inheritance of mitochondria in Chinese pine. Correspondence and reprints: State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Science, China Agricultural University, Beijing 100094, People’s Republic of China.     

侧金盏花双受精进程研究

应用荧光显微镜和常规石蜡切片观察侧金盏花(Adonis amurensis)花粉管生长和受精作用的全过程。结果表明,侧金盏花为湿型柱头,授粉后1–2小时,花粉粒与柱头识别;授粉后2–4小时,花粉粒萌发;授粉后4–6小时,花粉管进入柱头。侧金盏花的受精模式为珠孔受精,授粉后10小时,精子被释放;授粉后30小时,精核与卵核融合;授粉后7天合子形成;授粉后15天合子进入分裂期,合子休眠期为8天。2个极核在受精前不融合,授粉后14–16小时,精核与1个极核融合;授粉后20–22小时,受精极核与另1个极核融合形成初生胚乳核。双受精作用属于有丝分裂前配子融合型。通过实验确定了侧金盏花受精过程的雌雄性细胞融合形态变化与相应经历的时间及其合子休眠期。研究结果丰富了侧金盏花胚胎学资料,对其今后的育种及转基因研究具有重要意义。