Western white pine (Pinus monticola Dougl.) reproduction: I. Gametophyte development

Male and female gametophyte development are described from light and transmission electron microscope preparations of ovules from first and second year Pinus monticola Dougl. seed cones. In the first year of development, pollen tubes penetrate about one-third the distance through the nucellus. The generative cell and tube nucleus move into the pollen tube. The megagametophyte undergoes early free nuclear division. First-year seed cones and pollen tubes become dormant in mid-July. In the second year, seed cones and pollen tubes resume development in April and the pollen tubes grow to the megagametophyte by mid-June. Early in June the generative cell undergoes mitosis, forming two equal-size sperm nuclei that remain within the generative cell cytoplasm. The generative cell has many extensions and abundant mitochondria and plastids. The megagametophyte resumes free nuclear division, then cell wall formation begins in early July. Cell wall formation and megagametophyte development follow the pattern found in other Pinaceae. Three to five archegonial initials form. The primary neck cell divides, forming one tier of neck cells. Jacket cells differentiate around each central cell. The central cell enlarges and becomes vacuolate; then vacuoles decrease in size and the cell divides, forming a small ventral canal cell and a large egg. Plastids in the central cell engulf large amounts of cytoplasm and enlarge. This process continues in the egg, and the peripheral cytoplasm of the egg becomes filled with transformed plastids. Mitochondria migrate around the nucleus, forming a perinuclear zone. The wide area of egg cytoplasm between these two zones has few organelles. A modified terminology for cells involved in microgametophyte development is recommended. Received: 9 December 1999 / Revision accepted: 30 April 2000     

黑节草双受精过程及胚胎发育的研究

黑节草从传粉到受精约需１３０ｄ,精子在花粉管中形成,胚囊发育属蓼型胚囊,因反足细胞较早退化,故受精前胚囊多只由卵器和中央细胞组成。精卵核融合时,精核染色质进入卵核后凝集成颗粒状,并在原位与卵核的染色质融合,雌、雄性核仁一直维持至合子的第一次分裂期前。双受精作用正常,属于有丝分裂前配子融合类型,初生胚乳核发生２－３次分裂后逐渐退化消失,胚的发育局限于球形胚阶段。     

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.     

樟子松受精作用和原胚的选择

樟子松发育成熟的雄配子体中的精子6月15日左右在颈卵器中上部与卵细胞结合,进行受精作用,其后,受精卵进行游离核分裂,形成8个子核时,开始形成细胞壁。它们再分裂1次,形成16个细胞的原胚。接着胚柄细胞层迅速生长、伸长,把下面的原胚送出颈卵器基部的细胞壁,进入胚乳中的溶蚀腔。原胚吸收溶蚀腔中的营养,生长发育。初期,胚的数目往往很多,但常常只有1个发育成熟。     

樟子松大孢子的发生和雌配子体的形成过程

樟子松大孢子母细胞经一系列变化,发育成雌配子体。在哈尔滨地区樟子松大孢子母细胞于每年6月8～14日形成,接着进行减数分裂,于6月16～20日形成大孢子。随着大孢子核的分裂,进入游离核时期,并于次年5月28日～6月4日形成细胞壁,幼雌配子体中出现颈卵器原始细胞,它分裂一次形成颈细胞和中央细胞。6月7～9日中央细胞分裂成卵细胞和腹沟细胞,6月13～15日颈卵器发育成熟。成熟的颈卵器含有颈细胞、腹沟细胞和卵细胞,但颈细胞和腹沟细胞已经退化。     

Fertilization and rejection of spermatozoids by egg cells in artificially pollinated ovules ofEncephalartos (Zamiaceae)

The structure and behaviour of free female, male and proembryonal nuclei ofEncephalartos villosus Lem. were studied during a light-microscopical investigation of serially sectioned archegonia in successfully pollinated ovules. Before spermatozoids were released from the pollen tubes into the archegonial chamber, the ventral canal nucleus had disintegrated in the neck region of the egg cell among minute, amoeboid bodies with PAS-positive granules. In archegonia containing multiple spermatozoids, the egg nucleus was unobtrusive and syngamy followed by proembryo formation regularly resulted. The egg cell usually reacted violently in archegonia penetrated by a single spermatozoid. These reactions were regarded as rejection phenomena and considered as indicators that the egg cell can differentiate between compatible and incompatible male gametes.     

Secretions from the female gametophyte and their role in spermatozoid induction in Cycas revoluta

In cycads, spermatozoids are released from pollen tubes and swim in fluid toward the archegonia. The source of this fluid was examined using Cycas revoluta Thunb. ovules placed in culture. Dissected female gametophytes just before fertilization produced copious fluid on their upper surface. The fluid first appeared around the archegonial chamber and then on the inside of the archegonial chamber. When this fluid was applied to dry turgid pollen tubes, they discharged spermatozoids 12 h later. The archegonial neck appeared as two semi-spherical swellings, whereas the four neck cells later became visible and they separated in a schizogenous manner. Many globose particles appear on the top of the archegonial neck cells when the fluid is present. The contents of pollen tubes, spermatozoids and surrounding liquid intermingle with the secreted fluid. The female gametophyte differs in ultrastructure during the stages before and after fluid secretion, the latter showing changes suggestive of fluid secretion from the female gametophyte.     

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.     

云南松雌雄配子体的发育

云南松(Pinus yunnanensis Fr.)雄配子体于10月在小孢子叶腹面产生二个小孢子囊,内有许多进行分裂的造孢组织细胞。第二年一月下旬至二月初小孢子母细胞进行减数分裂。在分裂期间,细胞内所贮存的淀粉粒的分布发生变化。二月初四分体小孢子形成,绒毡层细胞解体。二日中旬单核花粉粒形成,外壁扩展形成二个异极对称的气囊。三月花粉在四细胞时期散发。 雌配子体于二月上旬在珠心皮下分化出孢原细胞。二月下旬大孢子母细胞进入减数分裂期。三月初直列四分体大孢子形成,珠孔端三个退化,合点端一个功能大孢子进入有丝分裂期,形成约32个游离核的配子体。次年三月初雌配子体形成,四月初中央细胞核分裂,四月底颈卵器成熟,卵核周围产生辐射状原生质纤丝。五月初受精开始。云南松雌雄配子体的发育与亚热带分布的P.roburghii相似。     

银杏雌雄配子体发育及胚胎形成的研究进展

银杏(Ginkgo biloba)是现存最古老的裸子植物之一, 其生殖过程表现出许多原始特征和独特性状, 长期以来备受国内外专家的关注。经过近100年的研究取得了显著成果: (1) 银杏雄配子体发育周期长, 经历了从平周分裂到斜背式分裂,并最终垂周分裂形成带有鞭毛的游动精子; (2) 银杏雌配子体发育经历较长的游离核期和细胞化期, 分化形成颈卵器母细胞并经平周分裂、垂周分裂和斜向分裂形成成熟的颈卵器(包括有4个颈细胞和1个卵细胞); (3) 推测其精细胞中的液泡状结构为受精过程中的遗传物质载体; (4) 原胚的形成经历了游离核期和细胞化期。该文针对国内外最新银杏生殖生物学方面的研究进展, 对银杏雌雄配子体发育、受精过程以及胚胎形成等方面进行较为系统全面的分析和总结, 为进一步的银杏生殖生物学研究提供有价值的参考资料。     

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.     

On the Systematic Position of Amentotaxus from Its Embryological Investigation

The present paper deals with the embryological study and the systematic position of Amentotaxus argotaenia (Hance) Pilger. The material used was collected during 1980-1981 from Jin-fo Shan, 1400-1600 m, Sichuan Province, China. The species is dioecious. The male cone sheds its pollen during the period from the end of May to the middle of June. The pollen at mature stage is 2-celled. Pollen chamber appears obvious at the end of the nucellus. When pollen grains are dispersed, megaspore mother cell, which is situated deep in the nucellus, is in meiosis. The megaspore divides mitotically after pollination and the free nuclei of female gametophyte divide for the last time at the end of June. The wall formation takes place at the stage of 256 free nuclei. The development of archegonia takes place at the beginning of July and the fertilization occurs about July 20-23. The fertilized egg divides successively four times and results in a 16-nucleate proembryo. The young embryo is developing in August. It is interesting to note that the development of the young embryo is very slow. When the seed reaches the mature stage from June to July in the following year, the multicellular masses of the young embryos resulted from simple polyembryony remain immature within the female gametophyte. No cleavage polyembryony has been found. The subsequent embryogeny takes place after the seed has shed. Keng (1975) considers that Amentotaxus links the Taxaceae with Cephalotaxaceae. Our embryological data support Keng’s conclusion since they share (1) compound microstrobilus, (2) 2-celled pollen grains at shedding stage and (3) the rather long life cycle. Keng (1975) also mentions that Podocarpaceae may connect with Taxaceae through Phyllocladus. According to Keng the Podocarpaceae is related to Taxaceae to certain degree. It is obvious that the primitive spike-like male strobilus like the one in Cordaitales is obviously retained in Podocarpus spicatus and P. andinus of Podocarpaceae and Amentotaxus of Taxaceae. In addition, like in Amentotaxus there are 16 nuclei before wall formation in the proembryo of Podocarpus nivalis. These facts may well indicate that at least the Podocarpaceae and the Taxaceae were derived from a common stock. As far as the Taxaceae is concerned the authors tend to support the view of Koidzumi (1932) that Amentotaxus and Austrotaxus should be put in the same tribe since both possess the spike-like strobilus, the long life cycle and the seed maturation in the following year. They are probably rather primitive genera in the Taxaceae. The proembryogeny of Torreya is more or less specialized. It may be placed in a rather advanced tribe and the tribe Taxeae (including Taxus and Pseudotaxus)may be between the above two tribes. In conclusion, the Taxaceae is related to the Coniferales in certain respects and, as Keng (1975), Harri (1976) and Wang et al. (1979) have pointed out recently, placing of the Taxaceae in Coniferales is rather justifiable.     

FERTILIZATION IN BARLEY

The mature embryo sac of barley consists of an egg, two synergids, a central cell, and up to 100 antipodal cells. At shedding the male gametophyte is 3-celled, consisting of a vegetative cell with a large amount of starch and two sperms having PAS+ boundaries. Before pollination the nucleus and cytoplasm of each synergid appear normal. After pollination the nucleus and cytoplasm of one synergid undergo degeneration. The pollen tube grows along the surface of the integument of the ovule, passes through the micropyle, and enters the degenerate synergid through the filiform apparatus. The pollen tube discharges the vegetative nucleus, two cellular sperms, and a variable amount of starch into the degenerate synergid. Soon after deposition the sperms migrate by an unknown mechanism to the chalazal end of the degenerate synergid. Sperm nuclei then enter the cytoplasm of the egg and central cell, ultimately resulting in the formation of the zygote and primary endosperm nucleus, respectively. Sperm boundaries do not enter egg or central cell, but it was not possible to determine the fate of other sperm components. Degenerate vegetative and synergid nuclei remain in the synergid after fertilization, constituting what are considered to be X-bodies in barley. The second synergid degenerates during early embryogeny.     

Ricerche Embriologiche su Senecio Vulgaris L. var. Thyrrenus Fiori

Abstract

Embryological researches on SENECIO VULGARIS L. var. THYRRENUS Fiori. — Male gametophyte, development of tapetal cells and female gametophyte have been studied in Senecio vulgaris L. var thyrrenus Fiori.

1) The development of male gametophyte is normal. Divisions of the microspore mother cells are of the simultaneous type. The division of the generative nucleus has never been observed till the pollen grain was in the anther.

2) The tapetal cells follow a very simple development. The nucleus of each cell divides only twice starting at the same time with the meiotic divisions of pollen mother cells but ending much earlier; subsequently, as usually happens with the Asteraceae, the ameboid involution of the tapetum begins. Endomitosis or any other process which leads to a polyploidy not due to nuclear fusion, has never been observed.

3) The female gametophyte is eight nucleate of the normal type (Polygonum). At maturity it shows only three antipodal cells whose nucleus undergoes at first, two or three divisions. Only later these new nuclei, always within the antipodal cell, may fuse in a polyvalent one.     

乌毛蕨配子体发育的研究

采用混合土培养乌毛蕨（Blechnum orientale）孢子,显微镜下观察记录其孢子萌发及配子体发育过程。结果表明：孢子黑褐色,赤道而豆形,极而观椭圆形,单裂缝。播种1周左右孢子萌发,萌发类型为书带蕨型,配子体发育为叉蕨型。丝状体5—10细胞时开始发育为片状体。播种2周后发育形成幼原叶体,成熟原叶体呈心脏形。原叶体边缘及表面均可产生毛状体,数量丰富,为单细胞。播种后1个月左右开始有颈卵器出现,成熟颈卵器颈部由4列细胞组成,3—5层细胞高。精子器产,扛时间较颈卵器早10d左右,精子器近圆球形,由3细胞组成。精卵受精后2周左右即可观察到从原叶体上生成的幼胚。     

Early Embryogeny and Its Starch Distribution in Amentotaxus

The present paper deals with the early embryogeny of Amentotaxus argotacnia (Hance) Pitger and its variation in starch distribution. Amentotaxus is endemie to China. The proembryos of Amentotaxus occurred at the end of July to early August, 1980–1981, When the zygote has sueeessive]y divided for four times, the daughter nuelei are becoming smaller and smaller after each division. At first, the zygote is 80–100 μ in diameter. Then, tbe free nuclei of the proembryo are 50–70 μ in diameter in two-nucleate stage, 36–50 μ in four-nucleate stage, and 29–32 μ in eight- to sixteen-nueleate stage, respectively. The wall formation of proembryos in Amentotaxas as in most other members of Taxaeeae also takes place at 16-nucleate stage. After wall formation the cells u each proembryo are arranged in two groups, upper one constitutes the cells of open tier (O) and the lower one, the primary embryonic eells (PE). The ratio of 0 to PE is 9:7 or 8:8 in some eases. The cells of open tier elongate and divide to form the ceils of upper tier (U) and the prosuspensor eens (S). In such ease, The ratio is U:S:PE= 8:8:8. When the eells of open tier and primary embryonie tier sometimes divide simultaneously, the primary embryo cells result in the embryo cells (E), and the ratio is U:S:E=9:9:14 or U:S:E=8:8:16. The young embryos of Amentotaxus begin to differentiate in the first week of August in Jin-Foshan (Golden Buddha Mountain), 1400 to 1600m. Sichuan Province, China The developmental features of the young embryo in Amentotaxus are as follows: (1) The development of the young embryos lasts for 10 to 12 months. This is very unique in Gymnosperms. The development of tile embryo in Amentotaxus is in some deg ree similar to that of Ginkgo, beeause their young embryos develop in maternal plants, whereas the late embryogeny takes place after shedding of the seed. (2) The young embryos pass through the winter at multieellular stage and the late embryos are still undifferentiated when the arils are getting red and the seeds begin to shed. It is interesting to note that the development of embryos are still staying at embryo seleetion stage of simple polyembryony when seeds were stored for six months in 15-20℃. As far as our information goes, the embryos in seeds should get over another winter until embryo matures. The embryos in Amentotaxus is un;quo in this respect and it is considered to be primitive. Though the seeds of pteridosperms have large female gametophytcs, none of embryos have been found in their fossil seeds. Probably their embryos are not well developed when the seeds mature and shed. Thus the embryos of fossil seeds are not easily preserved (Cronquist, 1968). The condition of embryonic developaleut in Amentotaxus resembles strongly with that in pteridosperms. From above, Amentataxus eould be the most primitive genus in Taxaceae. (3) Simple polyembryony in Amentotaxus is pronfinent. The prosuspensors sometimes divide to give rise to “suspensor embryos”. The general tendency of the starch distribution in male and female gametophytes is that the main regions of stareh are gradually transferred frmn mieropylar to chalazal end wiht the development of the ovule. After pollen germination the stareh proceeds together with the sterile cell, tube nueleus and spermatogenous cell down the arehegonium. In early developmental stage of female gametophyte, the starch region always appears round the upper part of the archegonia; after fertilization they mainly appear in tissue of female gamctophyte near the proembryo or young embryo to form the pyramidal region. It is worthy to note that the starch grains of male gametophyte are larger in size than those of female gametophyte and the former is much less in nmnber than the latter. So far as the embryo proper is concerned the starch grains densely appear armmd the nuclei of the embryos, espeeially those of the prosuspensors. Besides, near the basal part of aril and integument there is a stable region of polysaeeharide which shows the positive reaetion for PAS. This region is always present from the origin of aril to its mature and is an important feature of the ovule and seed in Amentotaxus,     

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.     

The early embryogeny of the genus Fokienia with a note on its systematic position

The present investigation deals with the early embryogeny of Fokienia hodginssii (Dunn) Henry et Thomas with a note on its systematic position. The material was collected on April 17 to September 13 in 1964 from longquan, Fengyang Shan in the province of Zhejiang, at alt. 1000 to 1400 M. Fokienia Henry et Thomas, a member of the subfamily Cupressoideae of northern hemisphere, is a monotypic genus with the only species Fokienia hodginssii (Dunn) Henry et Thomas. The zygote divides three times, giving rise to eight free nuclei, then wall-formation follows. As a result, the 8 cells of the proembryo are arranged in two groups: The upper one, the open tier (O) and the lower one, the primary embryo cells (PE). The relative number of these cells (0: PE) is usually 4:4, occasionally 5:3, rarely 6:2. The cells in the upper and lower groups divide simultaneously. A proembryo of three groups of cells may be formed. The upper tier (U), the suspensor tier (P), and the embryo cells (E). U:S:E is usually 4:4:8, occasionally 5:5:6. rarely 6:6:4. The U and E are of common in origin. The primary embryo cells sometimes remain undivided though the cells of upper tier divide as usual and the prosuspensor celts elongate also. In this case, U: S: E is 4:4:4 or 5:5:3 or 6:6:2. Cleavage polyembryony occurs quite often. Generally, the cleavage polyembryony is caused by the different growth rate of the primary suspensor. Sometimes, the terminal cells cut off when cells of primary suspensor are elongating. The terminal cells elongate and divide repeatly, thus a number of successive suspensor tubes are produced.This is a derivative type in the cleavage polyembryony of Fokicnia. This specialized type of polyembryony likes that of the Juniperoid. Different view points exist in the taxonomic treatments of the Cupressaceae. Many taxonomists divide the family Cupressaceae into 2—4 subfamilies. From the view point of the early embryogeny, the author considers that Li’s (1953) treatment is a more appropriate one. According to cone structure and arrangement of ovuliferous scales. Li (1953) divides the family Cupressaceae into two subfamilies; i.e. subfamily Cupressoideae of northern hemisphere having ovuliferous scales of imbricate arrangement and Callitroideae of southern hemisphere having scales of valvate arrangement. It is interesting to note that the wall-formation of proembryo in northern hemisphere plants of Cupressaceae takes place at 8 free nuclear stage, while those in southern hemisphere ones, at 4 free nuclear stage. Apparently, the status of proembryogeny gives support to the view points of Li (1953). From the point of view of early embryogeny,however, there are still more questions to be discussed. For example, in the subfamily Cupressoideae of northern hemisphere, Li considered the tribe Cupresseae as the primitive and the genus Thujopsis of the tribe Thujopsideae derives from genus Fokienia of the tribe Cupresseae. According to the data obtained from the early embryogeny, the author considers the tribe Thujopsideae to be the most primitive of the three tribes in the subfamily Cupressoideae, then the tribes Cupresseae and finally, the tribe Junipereae. Embryogenesis of Fokienia, the northern hemisphere members of the Cupressaceae, is a specific type, whose systematic position is possibly between Chamaecyparis andSabina.     

Isolation of Viable Male and Female Gametes in Cunninghamia lanceolata

Viable male and female gametes were isolated from pollinated ovules of Cunninghamia lanceolata (Lamb.) Hook. Prior to their penetration into the female gametophyte, the pollen tubes were drawn out from the nucetlus. The isolated pollen tubes were branched and one of them became swollen. An enlarged spermatogenous cell and subsquently a pair of sperm cells were formed as the pollen tube reached the regions over and against the archegonia. The sperm cells were released from the pollen tubes manually with the use of a stereomicroscope. The positive FDA reaction gave evidence of the sperm cells viability, and the Fluorescent Brightener 28 positive demonstrated the presence of cell wall. The egg cells were enzymatically isolated from the female gametophytes. The isolated egg cells were spherical, contained 1 to 2 large and many small vacuoles. FDA test showed the egg cells were viable, and the viability sustained for 8 days in 2 to 4 ℃ without any protectants. Fusion between single pair of male and female gametic protoplasts was attempted with PEG method, but only adhesion of the two was obtained.     

烟草雌、雄配子DNA 含量变化

采用显微分光光度法测定了烟草( Nicotiana tabacum) 精细胞和卵细胞的DNA 含量。烟草是二胞花粉, 花粉萌发后生殖细胞在花粉管中分裂形成精细胞。授粉后45 h 花粉管到达子房, 在花粉管内的精细胞DNA 含量为1C。当花粉管在退化助细胞中破裂, 释放出的两个精细胞开始合成DNA。在与卵细胞融合前,两个精细胞DNA 含量接近2C。随着精细胞的到达及合成DNA, 卵细胞也开始合成DNA, 融合前的卵细胞DNA 含量也接近2C。精、卵细胞融合后, 合子DNA 含量为4C。烟草雌、雄配子是在细胞周期的G2 期发生融合, 属于G2 型。