濒危植物南方红豆杉大孢子发生和雌配子体发育

南方红豆杉(Taxus chinensis var.mairei)为第三纪孑遗树种,也是我国Ⅰ级保护植物。本文通过对其大孢子发生及雌配子发育过程进行细胞学观察发现,南方红豆杉胚珠多为直立单生,偶见1个苞片内着生2个胚珠;大孢子母细胞减数分裂形成4个大孢子,其中3个退化,仅位于合点端的大孢子发育为功能性大孢子;功能性大孢子经过有丝分裂形成256个游离核;颈卵器单生,2~6个;从传粉到受精约2个月;有16个原胚自由核,原胚为标准型;简单多胚和裂生多胚并存,胚发育不同步。因南方红豆杉胚珠直立单生、游离核数目、传粉到受精间隔期、原胚游离核数目等特征与松科差异较大,而与广义柏科具有许多相似之处,故我们支持将红豆杉科置于广义柏科之下的观点。此外,本研究结果还表明,南方红豆杉大孢子发生与雌配子体发育正常,不是致其濒危的主要原因。     

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。     

ONTOGENY OF THE PALEOZOIC OVULE,CALLOSPERMARION PUSILLUM

The ontogeny of the upper Pennsylvanian age gymnospermous ovule, Callospermarion pusillum, is described from petrifaction specimens collected at the Berryville locality in Illinois. Ovules exhibit a wide range of dimensional and structural features that indicate an extensive developmental sequence. Specimens range from ovules with indistinct zonation of the thin-walled integument to those with thick-walled cells of the sclerotesta. The apex of the fleshy nucellus in some specimens is preserved as a cellular mound, while in others a well-formed cellular pollen chamber is present; still other ovules are characterized by a papery-thin nucellus and pollen chamber wall. The megagametophyte of most specimens is represented by a hollow megaspore membrane that may be restricted to the base of the nucellus, or occupy the entire seed cavity. In a few specimens cellular gametophytes are preserved, and in one ovule archegonia with supposed eggs are also present. Variability in each of the features is compared with ontogenetic changes in comparable structures of living gymnospermous ovules and is correlated with ovule size. A developmental sequence for the fossil ovules is proposed.     

极小种群野生植物崖柏的生殖物候、传粉及胚胎发育研究

以濒危植物崖柏（Thuja sutchuenensis Franch.）为对象,对其生殖物候、传粉机制进行观察,并采用石蜡切片法对其胚胎发育过程进行研究。结果显示：崖柏于8月分化出大、小孢子叶球,次年3月传粉,为花粉无气囊、具传粉滴、胚珠直立型传粉机制,球果于10月开裂;显微观察发现,传粉期花粉进入珠孔后,贮藏在珠心上方的贮粉室内,同时珠心组织中分化出孢原细胞,进入雌配子体发育阶段,5月中旬,花粉管开始萌发,6月初完成受精,进入胚胎发育阶段,10月初,胚胎发育成熟。研究表明崖柏从大、小孢子叶球形成至种子成熟的整个发育过程中均存在败育,而胚珠败育及雌配子体游离核时期至幼胚发育期间的败育是其生殖障碍的主要原因。本研究获得了崖柏生殖生物学的基础资料,为其人工繁育和制定保护策略提供了重要依据。     

杉木大孢子叶球发育过程中形态结构的变化

以福州生长的成年杉木（Cunninghamia lanceolata（Lamb.） Hook.）为实验材料,采用数码相机实地拍照、体视镜、半薄切片以及扫描电镜等方法,从形态学、解剖学系统观察了杉木大孢子叶球的发育过程。结果显示,2011年10月底至11月初,杉木大孢子叶球形成,此时大孢子叶球呈绿色,体积较小;翌年3月中下旬,大孢子叶球成熟,进入传粉期,期间大孢子叶球经历了由绿变黄的颜色转变、体积增大以及苞片开张的过程。胚珠发育过程中,胚珠原基于1月上旬发生,1月中下旬珠被和珠心组织已分化形成;2月下旬,珠心组织继续发育,形态呈椭圆型,并在其上方形成贮粉室,周围的珠被组织继续生长包围珠心组织,形成珠孔道;3月初珠孔形成,开口达到最大,胚珠的体积继续增大;3月中下旬,胚珠珠孔处开始分泌传粉滴。授粉后,传粉滴消失,珠孔上方的组织停止生长,珠孔开口亦不再增大。研究结果表明杉木大孢子叶球从分化形成到发育成熟需要约5个月的时间,胚珠的形态结构经过长期演化形成了许多适应风媒传粉的结构特征。     

太白红杉雌配子体的形成、受精、胚胎发育及其系统学意义

太白红杉(Iarix chinensis Beissn)雌球花于7月中下旬开始分化,9月上旬至9月中旬形成大孢子母细咆,10月中旬,大孢子母细胞进入休眠期。翌年4月底至5月初解除休眠,大孢子母细胞进行减数分裂,于5月10日左右形成直列四分体,随后珠孔端的3个大孢子退化,合点端的1个大孢子进一步发育,成熟卵细胞于7月初受精;花粉管将内含物释放入卵细胞后在尾部形成浓密物质沉淀;受精后,合子被染色较深的新细胞质所包围,并发现存在多精入卵的现象。合子经过两次连续有丝分裂,产生4个游离核后伴随新细胞质一起移至合点端,接着同时进行一次有丝分裂产生8个核,并分成上下两层后形成细胞壁,但上层细胞顶部不形成细胞壁。原胚发育属于松型。在幼胚阶段,我们发现部分胚珠发育异常,其雌配子体有的变为半透明状,有的则干瘪萎缩。太白红杉具简单多胚和莲座胚。9月中旬,成熟胚形成,成熟胚具5～6枚子叶。太白红杉从雌球花花芽分化到胚胎成熟历时14个月。     

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     

矮沙冬青雌配子体及胚胎发育研究

矮沙冬青子房单心皮1室,边缘胎座,弯生胚珠,胚珠具双珠被、厚珠心。大孢子孢原细胞发生于珠心表皮下,大孢子母细胞减数分裂形成直线排列的四分体,合点端大孢子具功能,并按蓼型胚囊发育,雌配子体成熟于4月中旬。双受精后,胚乳发育为核型。在矮沙冬青大孢子发生、雌配子体和胚胎发育过程中未发现异常现象,因此认为矮沙冬青濒危不存在雌性生殖结构与发育过程异常的内在因素。     

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.     

小盐芥大孢子发生和雌配子体发育

本论文研究了小盐芥(Thellungiella halophila)大孢子发生和雌配子体发育过程及该阶段与花蕾、花、果实外部形态的相关性。结果如下: 小盐芥雌蕊由2心皮组成, 侧膜胎座, 每室胚珠多数, 弯生, 双珠被, 薄珠心。孢原细胞位于珠心表皮之下, 直接起大孢子母细胞的功能。大孢子四分体线形排列, 合点端大孢子为功能大孢子, 胚囊发育为蓼型。     

小盐芥大孢子发生和雌配子体发育

本论文研究了小盐芥（Thellungiella halophila）大孢子发生和雌配子体发育过程及该阶段与花蕾、花、果实外部形态的相关性。结果如下：小盐芥雌蕊由2心皮组成,侧膜胎座,每室胚珠多数,弯生,双珠被,薄珠心。孢原细胞位于珠心表皮之下,直接起大孢子母细胞的功能。大孢子四分体线形排列,合点端大孢子为功能大孢子,胚囊发育为蓼型。     

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.     

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.     

Pollen tubes enter neighbouring ovules by way of receptacle tissue,resulting in increased fruit-set in Sagittaria potamogetifolia Merr

Using fluorescence microscopy, deposition of pollen on stigmas and pollen tube growth in the gynoecium of Sagittaria potamogetifolia Merr., a monoecious species with an apocarpous gynoecium, were observed. The maximum rate of pollination averaged 83.9 +/- 4.7 %, and the number of pollen grains per stigma ranged from zero to 30. Pollen tubes grew through one stigma to the base of the ovary at almost the same speed, but generally only one of the pollen tubes then turned towards the ovule and finally entered the nucellus through the micropyle. The other pollen tubes grew through the ovary base and the receptacle tissue into ovules of adjacent carpels whose stigmas were not pollinated or which had been pollinated later. This phenomenon is termed pollen tube 'reallocation' by the authors. To verify the direct effect of the phenomenon on fruit set, artificial pollination experiments were conducted in which two or more pollen grains were placed onto only one stigma in each gynoecium; frequently more than one fruitlet was obtained from each flower treated. The reallocation of pollen tubes among pistils in the gynoecium could effect fertilization of ovules of unpollinated pistils and lead to an increase in sexual reproduction efficiency. It would, to some extent, also increase pollen tube competition among pistils of the whole gynoecium.     

小蓬草的胚胎学研究

对小蓬草(Conyza canadensis)大小孢子发生、雌雄配子体形成、受精、胚及胚乳发育过程进行了研究,主要结果如下:花药四室,药壁由表皮、药室内壁、中层和绒毡层组成.表皮退化;药室内壁宿存,细胞柱状伸长,纤维状加厚;中层细胞退化较早,在小孢子母细胞减数分裂开始时仅存残迹;绒毡层于小孢子母细胞减数第一次分裂前期开始原位变形退化,属于腺质型绒毡层;小孢子母细胞减数分裂为同时型,四分体的排列方式主要为四面体形和左右对称形;成熟花粉粒多为3细-胞花粉粒,偶见2细-胞花粉粒.子房下位,2心皮,1室,单胚珠,基生胎座;单珠被,薄珠心,倒生胚珠,具发达的珠被绒毡层.珠心表皮下分化出大孢子孢原细胞,孢原细胞直接发育为大孢子母细胞,大孢子母细胞减数分裂形成4个大孢子直线形排列,仅合点端的大孢子发育成功能大孢子母细胞,胚囊发育为蓼型.两个极核在受精前融合为次生核,珠孔受精.胚乳发育属于核型,胚胎发育为紫菀型;具胚乳吸器.     

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.     

小蓬草的胚胎学研究

对小蓬草（Conyzacanadensis）大小孢子发生、雌雄配子体形成、受精、胚及胚乳发育过程进行了研究,主要结果如下：花药四室,药壁由表皮、药室内壁、中层和绒毡层组成。表皮退化;药室内壁宿存,细胞柱状伸长,纤维状加厚;中层细胞退化较早,在小孢子母细胞减数分裂开始时仅存残迹;绒毡层于小孢子母细胞减数第一次分裂前期开始原位变形退化,属于腺质型绒毡层;小孢子母细胞减数分裂为同时型,四分体的排列方式主要为四面体形和左右对称形;成熟花粉粒多为3-细胞花粉粒,偶见2-细胞花粉粒。子房下位,2心皮,1室,单胚珠,基生胎座;单珠被,薄珠心,倒生胚珠,具发达的珠被绒毡层。珠心表皮下分化出大孢子孢原细胞,孢原细胞直接发育为大孢子母细胞,大孢子母细胞减数分裂形成4个大孢子直线形排列,仅合点端的大孢子发育成功能大孢子母细胞,胚囊发育为蓼型。两个极核在受精前融合为次生核,珠孔受精。胚乳发育属于核型,胚胎发育为紫菀型;具胚乳吸器。     

The origin of the ovule

Of several theories for the origin of the ovule advanced in this century and based largely on fossil evidence, the telomic concept is widely considered the most plausible. Its principal tenet is the evolution of the integument through fusion of sterile branches or telomes around a terminal megasporangium. The only point of agreement in these theories is that the entire nucellus is a megasporangium that retains a single megaspore and the endosporic female gametophyte. Their differences center on the origin of the integument. A new concept offered here on evidence from ovules of both Paleozoic and extant seed plants significantly alters the telomic theory. It proposes that the nucellus is a sporangiophore of stem origin that bears a terminal megasporangium; that at least some of the fused integumentary telomes were fertile; and that among all features cited to characterize ovules, the unique nature of the retained megaspore alone defines the seed habit. Changes in the seed plant megaspore that extended the period of nutrient absorption over the whole course of female gametophyte development, along with complex physiological changes in the nucellus, were probably achieved along a single phylogenetic line beginning in a Late Devonian population of progymnosperms. For such a combination of events to have occurred more than once is highly unlikely, and, therefore, a monophyletic origin for seed plants is proposed. Several primitive features in ovule structure, some not evidenced since the Lower Carboniferous Period, occur in a mutant form of Arabidopsis thaliana isolated from genetically transformed plants. Their recurrence provides additional support for the proposed concept of ovule origin and also suggests that the genetic mechanisms for expression of primitive features in advanced taxa could be initiated in each case by mutation of a single homeotic gene.     

Studies on the Development of Male and Female Gametophytes in Sinojakia xylocarpa

Observed in this paper was the development of the microspore and megaspore, male and female gametophytes in Sinojakia xylocarpa, which is endemic to China. The anther comprises four microsporangia. Microspore wall forms simultaneously after meiotic division in PMCs. The arrangment of microspore in a tetrad is tetrahedral. Bicel lular pollen grains appear at the shedding stage. ‘They are 3-colporate, with irregular min ute-faveolate exine sculpture. The anther wall development is of the dicotyledonous type, and its endothecitum develops slight fibrous thickenings, which also form on some epidermal cells. The tapetum is glandular. The pistil with hollow style is composed of three carpels, and its ovary contains several anatropous ovules. The ovule is unitegmic, tenuinucellar, but no obturator was observed. The archesporial cell functions directly as the megaspore mother cell which forms a linear tetrad, but T-shaped tetrad was found in a few ovules. A Polygonum type embryo sac forms from the functional chalazal megaspore. In the mature embryo sac, the synergids are elongate with a large vacuole at the chalazal end, but the distrihution of vacuoles in the egg cell appears random. Two polar nuclei remain in contact with each other for a spell before the fertilization and the 3 antipodal cells may persist into early postfertilzation stages. Numerous starch gra ins occur in the embryo sac. According to the present embryological studies on Sinojakia xylocarpa and the works on embryogenesis by some early embryologist, authors consider that Styracaceae, Symplocaceae, Sapotaceae and Ebenaceae are rather closely related, and we alsoconsider it reasonable to put the 4 families mentioned above in Ebenales.     

The evolution of fertilization modes independent of the micropyle in Fagales and `pseudoporogamy'

In contrast to a majority of angiosperms showing porogamous fertilization, several fagalean families such as Betulaceae and Casuarinaceae are known to show chalazogamy, where fertilization is effected by a pollen tube passing through the chalaza instead of the micropyle. Our developmental study of pollen-tube growth in pistils of Myrica rubra (Myricaceae, Fagales) further shows that pollen tubes reached the nucellus before the micropyle is formed by the integument. Since fertilized ovules appeared as if the pollen tube had passed through the micropyle for fertilization, we propose the new term `pseudoporogamy' to this mode. By mapping diverse modes of fertilization, dependent or independent of the micropyle, onto a phylogenetic tree of Fagales, it appears that fertilization mode evolved from porogamy to chalazogamy and then further from chalazogamy to pseudoporogamy. Possible reasons for the evolution of fertilization modes independent of the micropyle in Fagales are discussed.