龙须草无融合生殖的胚胎学证据

采用石蜡切片技术对龙须草(Eulaliopsisbinata(Rotz)C.E.Hubb)进行了系统的胚胎学研究,证明龙须草为禾本科植物中一种新的无融合生殖材料。龙须草无融合生殖方式为无孢子生殖,在胚珠发育早期,多个珠心细胞特化为无孢子生殖原始细胞,由原始细胞发育为单核胚囊,经两次有丝分裂形成4核胚囊,进一步分化形成两种类型的成熟胚囊:(1)具1个卵细胞,1个助细胞和2个极核,占观察总数的67.6%;(2)具1个卵细胞,2个助细胞和1个极核,占观察总数的32.4%。胚囊发育属大黍型。多个无孢子生殖原始细胞可以同时发育,最后形成2个或多个胚囊,其比例为17.7%。胚珠内没有有性胚囊的发育。胚的发生有两种类型:(1)早发生胚(74%),开花前1~2d,极核未分裂前卵细胞分裂形成胚;(2)迟发生胚(26%),开花后2~3d,极核分裂形成多个胚乳游离核后,卵细胞启动分裂形成胚。存在多胚现象,多胚来自不同胚囊内卵细胞的孤雌生殖,多胚发生率为13%。胚乳由极核不经受精自发分裂产生。     

龙须草无融合生殖的胚胎学证据

采用石蜡切片技术对龙须草(Eulaliopsis binata(Rotz)C.E.Hubb)进行了系统的胚胎学研究,证明龙须草为禾本科植物中一种新的无融合生殖材料.龙须草无融合生殖方式为无孢子生殖,在胚珠发育早期,多个珠心细胞特化为无孢子生殖原始细胞,由原始细胞发育为单核胚囊,经两次有丝分裂形成4核胚囊,进一步分化形成两种类型的成熟胚囊:(1)具1个卵细胞,1个助细胞和2个极核,占观察总数的67.6%;(2)具1个卵细胞,2个助细胞和1个极核,占观察总数的32.4%.胚囊发育属大黍型.多个无孢子生殖原始细胞可以同时发育,最后形成2个或多个胚囊,其比例为17.7%.胚珠内没有有性胚囊的发育.胚的发生有两种类型:(1)早发生胚(74%),开花前1～2 d,极核未分裂前卵细胞分裂形成胚;(2)迟发生胚(26%),开花后2～3 d,极核分裂形成多个胚乳游离核后,卵细胞启动分裂形成胚.存在多胚现象,多胚来自不同胚囊内卵细胞的孤雌生殖,多胚发生率为13%.胚乳由极核不经受精自发分裂产生.     

草地早熟禾胚胎学研究 Ⅲ．多胚囊及多胚现象

报道了草地早熟禾中多胚囊的起源、发育和结构。在１个胚珠中,大孢子母细胞周围可以有一到多个起源于珠心细胞的胚囊原始细胞,并可以发育成为多胚囊,其中具有两个胚囊的可以发育成为成熟胚囊。起源于珠心的体细胞无孢子生殖胚囊的发育属于山柳菊型。两个成熟胚囊中,都可以形成胚和胚乳,因而形成了具假多胚的种子。位于中部的胚来源于珠心还囊,属于无融合生殖形成的胚。两个以上的多胚囊不能形成成熟胚囊。     

雾灵山草地早熟禾多胚囊和多胚现象的研究

无融合生殖是指未经精卵融合而产生后代的特殊生殖方式,它可以分为单倍体无融合生殖和二倍体无融合生殖;对于作物改良意义更大的是二倍体无融合生殖。多胚囊和多胚现象SHI是无融合生殖的表现形式。本文运用石蜡切片法、子房整体透明法研究了雾灵山草地早熟禾〖WTBX〗（Poa pratensis〖WTBZ〗 L.）多胚囊和多胚现象。结果表明,（1）草地早熟禾多胚囊来源有两种：一是来自大孢子母细胞,二是来自珠心细胞;（2）草地早熟禾多胚来源有四个：其一是有性生殖胚,其二是孤雌生殖胚,其三是无配子生殖胚,其四是珠心胚。     

黄花油点草胚胎发育研究

利用石蜡切片技术对百合科植物黄花油点草[Tricyrtis maculata(D.Don)Machride]双受精、胚及胚乳发育进行了研究,以明确其胚胎发育的特征,为百合科植物的系统研究提供生殖生物学资料。结果表明:(1)黄花油点草为珠孔受精;进入胚囊的2枚精子分别与卵细胞和中央细胞进行正常的双受精,其受精作用属有丝分裂前型。(2)受精后的初生胚乳核立即分裂,其发育方式为核型胚乳;早期的游离胚乳核沿胚囊的边缘分布,胚囊中央部位主要为胚乳细胞质,随着游离胚乳核数量的增加,胚乳核慢慢充满整个胚囊;当发育至球形胚早期阶段,在各胚乳核周围产生胚乳细胞壁,形成完整的胚乳细胞。(3)合子有较长的休眠时间,胚的发育方式为茄型;合子第一次有丝分裂为横裂,分裂后形成基细胞和顶细胞;基细胞经过3次横裂,形成一列胚柄细胞;顶细胞经过分裂形成胚体,依次形成球形胚、棒状胚和盾形胚。(4)种子成熟时胚无器官分化;成熟种子由种皮、胚和胚乳三部分组成。     

草地早熟禾假受精现象的研究

本文运用石蜡切片法、整体透明法研究了草地早熟禾的假受精现象。结果表明,草地早熟禾在两极核清晰可见时,卵细胞就已自发分裂成原胚,此现象占观察总数88％;但是,其极核必需经过受精过程才能发育为胚乳。     

草地早熟禾假受精现象的研究

本文运用石蜡切片法、整体透明法研究了草地早熟禾的假受精现象。结果表明,草地早熟禾在两极核清晰可见时,卵细胞就已自发分裂成原胚,此现象占观察总数８８％;但是,其极核必需经过受精过程才能发育为胚乳。     

冠果草的胚胎学研究

冠果草花药壁的发育为单子口十型,绒毡层为周原质团型。小孢子母细胞减数分裂为连续型,四分体呈左右对称式排列,成熟花粉为三细胞型。双珠被,假厚珠心,倒生胚珠。胚囊发育为葱型,成熟胚囊的特点是两个极核分别位于中央细胞两端,不融合成次生核。受精过程中,一个精于与卵核融合形成合子,另一精子先与珠孔端极核融合,之后受精极核再移动到合点端与另一极核融合,形成初生胚乳核。胚的发育为石竹型。成熟胚呈马蹄形,具有2片真叶。胚乳发育为沼生目型。随着胚的发育,胚乳细胞逐渐解体,成熟种子中无胚乳。     

濒危植物——长喙毛茛泽泻的受精作用及胚和胚乳的发育

长喙毛茛泽泻Ranalisma rostratum Stapf的配子融合开始较早,但精核与极核融合速度较快。精核与珠孔极核融合后即移向胚囊合点端与另一极核再融合形成初生胚乳核。初生胚乳核第一次分裂后形成两个细胞,以后珠孔端胚乳细胞再进行游离核分裂,胚乳发育属于沼生目型。开花后两天,合子分裂,八分体呈4层,每层呈两个细胞方式排列胚胎发生属于石竹型。     

四倍体双穗雀稗兼性无孢子生殖的研究

研究了四倍体双穗雀稗(Paspalum distichum L)无孢子生殖胚囊、胚胎发育以及假受精特点。当其大孢子母细胞发育至四分体阶段时,大多数情况下会发生四分体退化,同时有多个特化珠心细胞发育为1—3个无孢子生殖胚囊的现象。成熟无孢子生殖胚囊一般3核,包括1个卵细胞和2个极核。卵细胞在抽穗前就能自发分裂形成原胚团,而极核则在抽穗和传粉后参与假受精形成胚乳。当胚珠内存在多个无孢子生殖胚囊时,只是靠近珠孔端的1个无孢子生殖胚囊内的极核与精核结合,而其它的并不参与。种子成熟后出现很低频率的二胚苗。此外,还能观察到少量的有性生殖胚囊的发育以及有性生殖胚囊和无孢子生殖胚囊在同一胚珠中的发育现象,因此判断该类群为兼性无孢子生殖体。     

Embryological problems in the apomictic speciesChondrilla juncea L. (Compositae)

Examination of the embryo and endosperm development in triploidChondrilla juncea L. (2n=15) from Poland confirmed the occurrence of autonomous apomixis in this species. Numerous degenerated embryos were formed which might be one of the factors which increased the observed seed sterility. In addition, twin embryos were often found at young developmental stages and in germinating seeds. Endosperm developed from polar nuclei or from the secondary nucleus. These processes have been proved by counting chromosome numbers in early developmental stages of the endosperm. The problems of the prevailing type of endosperm (nuclear or cellular), the correlation of embryo and endosperm development and the period of cellularization of nuclear endosperm have remained unsolved.     

The occurrence of phenotypically complementary apomixis-recombinants in crosses between sexual and apomictic dandelions (<Emphasis Type="Italic">Taraxacum officinale</Emphasis>)

Apomictic seed development in dandelion ( Taraxacum officinale) involves (1) restitutional meiosis (diplospory), (2) egg cell parthenogenesis, and (3) autonomous endosperm development. The question is whether these elements of apomixis are controlled by one single gene or by several independent genes. Five triploid non-apomictic hybrids, obtained in diploid sexual × triploid apomict crosses were characterized using cyto-embryological and genetic methods. Nomarski-differential interference contrast microscopy and the transmission of microsatellite markers and ploidy levels indicated that the hybrids combined elements of the apomictic and the sexual developmental pathway. Hybrids form two complementary groups with respect to the presence or absence of parthenogenesis and autonomous endosperm development. The occurrence of complementary apomixis-recombinants suggests that parthenogenesis and autonomous endosperm development in Taraxacum are regulated independently by different genes. This study also indicates that early embryo development is independent of endosperm formation, but that endosperm is essential for later embryo growth.     

Genetic diversity and reproductive biology in ecotypes of the facultative apomict Hypericum perforatum L

Apomixis is a mode of asexual reproduction through seed. Progeny produced by apomixis are clonal replicas of a mother plant. The essential feature of apomixis is that embryo sacs and embryos are produced in ovules without meiotic reduction or egg cell fertilisation. Thus, apomixis fixes successful gene combinations and propagates high fitness genotypes across generations. A more profound knowledge of the mechanisms that regulate reproductive events in plants would contribute fundamentally to understanding the evolution and genetic control of apomixis. Molecular markers were used to determine levels of genetic variation within and relationship among ecotypes of the facultative apomict Hypericum perforatum L. (2n = 4x = 32). All ecotypes were polyclonal, being not dominated by a single genotype, and characterised by different levels of differentiation among multilocus genotypes. Flow cytometric analysis of seeds indicated that all ecotypes were facultatively apomictic, with varying degrees of apomixis and sexuality. Seeds set by haploid parthenogenesis and/or by fertilisation of aposporic egg cells were detected in most populations. The occurrence of both dihaploids and hexaploids indicates that apospory and parthenogenesis may be developmentally uncoupled and supports two distinct genetic factors controlling apospory and parthenogenesis in this species. Cyto-embryological analysis showed that meiotic and aposporic processes do initiate within the same ovule: the aposporic initial often appeared evident at the time of megaspore mother cell differentiation. Our observations suggest that the egg cell exists in an active metabolic state before pollination, and that its parthenogenetic activation leading to embryo formation may occur before fertilisation and endosperm initiation.     

Apomixis in <Emphasis Type="Italic">Eulaliopsis binata</Emphasis>: characterization of reproductive mode and endosperm development

Apomixis represents an alteration of classical sexual plant reproduction to produce seeds with essentially clonal embryos, stimulating wide interest from biologists and plant breeders for its ability to fix heterosis. Eulaliopsis binata (Poaceae), is identified here as a new apomictic species. Embryological investigation indicates that the developmental pattern of embryo sac formation in E. binata represents gametophytic apospory, the embryo originating from an unreduced cell, without fertilization and the mode of endosperm development was autonomous. Sexual embryo sacs were found with a frequency of 1–4% depending on the biotype. The DNA content of nuclei (C-value) in mature seeds was screened by flow cytometry (FCSS) and demonstrated that the endosperm was derived autonomously without fertilization and the three biotypes of E. binata showed varying degrees of apomixis. The Wide-leaf type showed obligate apomixis whereas the Slender-leaf and the Red-haulm type displayed facultative apomixis. In addition, adventitious embryos were observed on the wall of ovary, integument and nucellus cells, indicating that E. binata produces embryos via a mixture of apospory and adventitious embryony.     

花椒和野花椒的无融合生殖

花椒与野花椒的胚囊发育类型属蓼型,成熟胚囊的卵器退化。花椒无雄花,不发生双受精,自发形成胚乳并产生珠心胚。野花椒虽有正常花粉,人工授粉后能萌发,但在花粉管长入胚囊之前卵器已解体,中央细胞中已形成胚乳游离核,因此也不发生双受精,由珠心细胞自发形成胚。这种现象是花椒和野花椒在长期进化过程中形成的一种十分特化的适应。     

Tripsacum dactyloides (Poaceae): a natural model system to study parthenogenesis

Diplosporous apomeiosis, formation of unreduced embryo sacs primarily of the Antennaria type, followed by parthenogenetic embryo development and pseudogamy (fertilization of the central cell) describe gametophytic apomixis within the Tripsacum agamic complex. Tripsacum dactyloides (Eastern gamagrass) is a close relative of domesticated maize and was chosen as a natural model system to investigate gene expression patterns associated with parthenogenesis. The genome size of diploid sexual and polyploid apomictic T. dactyloides was estimated by flow cytometry to be 7.37 pg (2C), 14.74 pg (4C) and 22.39 pg (6C), respectively. The diploid genome size is thus approximately 1.352 larger than that of maize. The apomeiotic-pseudogamous pathway of seed formation was demonstrated at a rate of 92% by the flow cytometric seed screen (FCSS) with single mature seeds in tetraploid accessions. This number includes twin embryos which were detected in 13% of the seeds analyzed. Fertilization of unreduced egg cells (B_III hybrids) was measured in 10% of apomictic seeds. Autonomous (fertilization-independent) embryo development and fertilization-dependent endosperm formation were confirmed by pollination of tetraploid T. dactyloides with a diploid transgenic maize line carrying an actin::#-glucuronidase (GUS) reporter construct. GUS expression was detected after pollination in the developing endosperm, but not in the embryo. In similar intraspecific crossing experiments with maize, GUS expression was detected in both the embryo and endosperm. A protocol was established for microdissection of embryo sacs and early parthenogenetic embryos of T. dactyloides. Together, these techniques provide new tools for future studies aimed at comparing gene expression patterns between sexual maize and sexual or apomictic T. dactyloides.     

Apomixis and amphimixis in flowering plants

The objective of the present article is to compare apomictic and sexual reproduction (amphimixis) in flowering plants. Light-optical and ultrastructural aspects of the cytoembryological processes in apomicts, beginning with the early stages of development of the ovule and concluding with the newly formed seed, are considered. In the overwhelming majority of apomicts, an inability to develop an autonomous endosperm or to form viable seeds without the involvement of the process of fertilization of the nuclei of the central cell of the embryo sac is observed. Characteristic features of the ultrastructural differentiation of the megasporocytes in diplospory, of aposporous initial cells in apospory, of embryocytes in adventive embryony, and of ovicells in parthenogenesis and synergids in apogamety are identified and are compared to the generative structures of amphimicts. The hypothesis is made that the mechanisms of genetic regulation in the formation and development of the generative structures in apomixis and amphimixis are similar at the cellular level. The present study is not a survey of apomixis in general. Previously published original results that have been obtained by the present author as a result of many years of research in the area of apomixis have served as a basis for the preparation of the study.