提高水稻双胚苗和不定胚频率的研究

以水稻双胚苗中提高不定胚的发生频率是研究利用无融合生殖固定F_1代杂种优势的主要途径之一。通过特殊的选育方法,已将原有双胚苗品系的双苗率由30％提高到60％左右。由子不定胚每双苗的发生率有着一定的内在联系,随着双苗率的提高,内含不定胚的频率也有相应提高的趋势。又据细胞学观察,认为不定胚发生在远离珠孔端的部位,故在种子中部的萌发苗可能是不定胚产生的种子形态学特征。今年,我们采用特定苗位作标记的筛选方法,选取种子正中部萌发苗并结合对其它非正中苗的人工切除,使水稻的不定胚频率由原来的2.6-5.1％提高到21.2％。水稻不定胚频率的大幅度提高,为无融合生殖在杂交水稻育种中的研究应用开拓了诱人的前景。     

水稻双胚苗无融合生殖胚胎学研究初报

将子房整体染色透明观察后,通过筛选材料石蜡切片的方法,研究发现水稻双胚苗品系APⅣ中,存在低频率的无融合生殖。开花前珠心细胞特化并分裂发育形成不定胚。随着珠心组织的退化,不定胚长八胚囊腔中,以球形胚形式存在。开花后4天,球形胚开始分化。球形胚在胚囊中可以和合子胚并存,也可以单独存在。但双胚主要来自受精的助细胞或合子胚纵裂。开花后3—4天,在胚乳腔中着色深的球状结构是胚乳部份脱落形成的,不是不定胚。珠孔端反足细胞团不能形成胚。     

水稻“多胚苗”的形态学观察

“无融合多胚苗”水稻Ｃ１００１Ｂ和它的杂种Ｃ１００１Ｂ×Ｃ１００１Ａ 的未成熟颖果１５００ 粒;田间和人工萌发的单苗、双苗和三苗实生苗共２７０２ 株进行了形态解剖观察,结果如下：１．Ｃ１００１Ｂ和它的杂交颖果中只含一个胚;２． 由单个胚萌发的双苗和三苗幼苗是多苗实生苗。双苗实生苗的侧苗１ 和三苗实生苗的侧苗１ 是由主苗基部胚芽鞘的腋芽所形成,侧苗２由侧苗１ 基生叶的腋部发育而来。因此,多苗实生苗系同一个胚发育的主苗及其腋芽系统所组成;３． 颖果中胚的位置和朝向的变化和侧苗生长的两种类型,导致主苗与侧苗位置上的不同。至于已报道的水稻“多胚苗”形成的无融合生殖类型和推测尚有待提出胚胎学的论据。     

几种具无融合生殖特性的植物多胚和多苗现象的观察

报道6种具无融合生殖特性的植物种子的胚数和萌发实生苗数的观察结果。金桔(Fortunellamargarita (Lour.)Swingle)、蜜桔(Citrus unshiu Marcoritch)和花椒(Zanthoxylum bungeanum Maxim.)具珠心胚,含多胚种子频率分别为97.50％、100％和45.00％;多胚种子的胚数范围分别为2～49,3～54和2～6。草地早熟禾(Poa pratensis L.)、滨草(Elymus rectisetus)和湖北海棠(Malus hupehensis(pampon.)Rehd.)具非减数配子体无融合生殖特性,含多胚种子频率依次为34.25％、8.11％和37.50％;前两种的多胚种子中胚数范围为2～3,后者为2～15。蜜桔、草地早熟禾和湖北海棠种子萌发多苗的频率分别为22.00％、6.14％和2.22％。描述了多胚种子中胚的形态、位置和分布。对6种植物含单胚种子的胚的来源进行了分析。初步结论:1.具无融合生殖现象的植物种子含多胚和萌发多苗的特性可作为寻找具无融合生殖特性的植物的形态学指标;2.对其筛选的材料进行大孢子发生、胚囊形成和早期胚胎发育的研究,以期阐明多胚来源和生殖类型。     

多胚水稻ApⅢ（双13）的胚胎学观察

对多胚水稻（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）ＡｐⅢ的大量成熟颖果、人工萌发的幼苗和开花后３～５ ｄ 的幼嫩颖果进行的整体解剖和显微制片观察表明：ＡｐⅢ的５０００粒成熟颖果中,８９．０％ 含单胚单苗,８．９％ 和１．２％分别含双胚双苗和三胚三苗;７００多粒幼嫩颖果中,９０．０％ ～９５．０％ 含单胚,５．０％ ～７．０％ 含双胚。因制片的数目有限,未见到含三胚的;在含单胚和多胚颖果中,胚均位于同一胚囊的珠孔端,未见到胚囊以外存在不定胚。根据上述结果,似可以认为ＡｐⅢ单粒颖果的双胚和三胚是由同一胚囊内的卵细胞和１或２个助细胞受精或不受精发育而来的     

水稻HDAR细胞胚胎学研究─—不定胚的发生和发育

对水稻HDAR胚胎发育过程的进一步研究表明,水稻HDAR中有2．72％（17／434）的不定胚发生和发育。其不定胚起源于胚珠内的珠心细胞。不定胚起始细胞启动分裂时,胚囊发育至2核或4核时期,8核胚囊时期,胚珠内已形成多细胞不定胚结构。随后不定胚细胞不断分裂并逐渐挤进胚囊。开花传粉后,不定胚利用胚乳提供的营养可以继续发育和分化。不定胚可以和合子胚一起发育,有时合子胚败育,不定胚继续发育并分化。讨论了水稻HDAR中不定胚的发生,及其发生远早于合子胚的意义。     

水稻HDAR细胞胚胎学研究─—不定胚的发生和发育

对水稻ＨＤＡＲ胚胎发育过程的进一步研究表明,水稻ＨＤＡＲ中有２．７２％（１７／４３４）的不定胚发生和发育,其不定胚起源于胚珠内的球心细胞,不定胚起始细胞启动分裂时,胚囊发育至２核或４核时期,８核胚囊时期胚珠内已形成多细胞不定胚结构,随后不定胚细胞不断分裂并逐渐挤进胚囊,开花传粉后,不定胚利用胚乳提供的营养可以继续发育和分化,不定胚可以和合子胚一起发育,有时合子胚败育,不定胚继续发育并分化。讨论了水     

水稻“无融合多胚苗”的形态学观察

水稻“无融合多胚苗”的形态学观察母锡金,陈祖铿,王伏雄（中国科学院植物研究所,北京１０００４４）“无融合多胚苗”水稻,ＣｌｏｏｌＢ和它的杂种,ＣｌｏｏｌＢ×ＣｌｏｏｌＡ,未成熟颖果１５００粒,田间和人工萌发的单苗、双苗和三苗实生苗共２７０２株进行了形...     

多胚水稻品系SB—1的细胞胚胎学研究：助细胞无配子生殖

对多胚水稻（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）品系ＳＢ－１ 自然群体中单倍体来源进行的细胞胚胎学研究表明,卵器中的一个或两个助细胞能在受精之前发育成胚,其发生率分别为观察子房数的２．２４％ 和１．７５％ 。在１ 个助细胞提前发育成胚的胚囊中观察到双受精作用。在开花后３—８ ｄ的颖果中观察到１．６２％ 的与合子胚迥异的异位瘦长单胚和０．８１％ 的有胚无胚乳子房,根据胚所处的位置认为其来源于助细胞无配子生殖。此外,还观察到极少数的中央细胞未经受精自发产生胚乳的现象。讨论了ＳＢ－１中发生的无配子生殖的意义     

水稻“双－3”多胚发生的研究

叶秀,陈泽濂,黎垣庆 　　水稻ＭＩＶ（双－３、籼稻）传粉后可以有多个花粉管同时进入胚囊．大多数胚囊的合子发育为一个正常的胚,但是有少数合子胚发生裂生并分化形成双胚芽和一胚根．有些胚囊的助细胞和卵细胞同时受精后,分别发育为助细胞胚和合子胚;有些胚囊中的反足细胞团可直接发育为胚．可见“双－３”水稻除有正常合子胚外还存在助细胞胚和反足细胞匹的多胚现象．     

植物无融合生殖相关基因研究进展

无融合生殖是指不经过精卵融合即可形成胚从而进行种子繁殖后代的一种特殊的无性生殖方式, 无融合生殖胚的形成没有父本的参与, 其后代是母本基因型的完整克隆, 因此是植物杂种优势固定与利用的一种最理想的途径, 具有巨大的潜在利用价值, 被誉为“无性革命”。按其胚体发生的途径, 无融合生殖可分为二倍体孢子生殖、无配子生殖和不定胚生殖三种类型。本文介绍了植物胚发育、胚乳发育、减数分裂等涉及无融合生殖过程的相关基因的研究进展, 同时介绍了可能与植物无融合生殖途径调控相关的几个基因片段的研究情况。     

杂交柑桔的胚胎发育(英文)

柑桔类珠心胚现象对研究其进化和种质保存方面有着重要意义,但是给其有性杂交育种工作造成相当大的麻烦：它不仅使杂交后代难于区分合子苗和珠心苗,而且会使合子胚发育中途天折。解决的途径是适时分离合子胚进行单独培养。为此,本工作于柑桔胚胎发育的不同时期取样、固定、观察,研究其合子胚发育规律,确定珠心胚侵入胚囊的时期。结果表明：一般情况下,授粉后３０ｄ,合于处于单细胞状态（Ｆｉｇ．１Ａ）;授粉后４０ｄ;合子开始分裂（Ｆｉｇ．１Ｂ）;授粉后４５ｄ,合子胚形成８－１６个细胞,珠心胚开始分裂（Ｆｉｇ．１Ｃ）;授粉后５０ｄ,合子形成球形胚,珠心胚尚未侵入胚囊（Ｆｉｇ．１Ｄ＆Ｅ）;授粉后５５ｄ,合子胚为球形胚或早心形胚,开始有少数珠心胚侵入胚囊（Ｆｉｇ．１Ｄ＆Ｅ）;授粉后６０ｄ,合子胚为心形胚（Ｆｉｇ．１Ｇ）。授粉后８０ｄ,合子胚为晚心形,珠心胚巨大量侵入并迅速发育（Ｆｉｇ．１Ｈ）。杂交的组合对合子,胚珠及果实发育均有一定影响,主要取决于杂交亲本。与种间杂交相比,属间杂交其合子分裂较迟,合子胚发育较慢,珠心胚侵入时期也较迟（Ｔａｂｌｅｌ,Ｆｉｇ．２）。这种发育速度的差异,引起所产生的合子胚的大小（Ｔａｂｌｅ２）珠心胚侵率（Ｔａｂ     

Patterns of storage protein and triacylglycerol accumulation during loblolly pine somatic embryo maturation

Conifer somatic embryo germination and early seedling growth are fundamentally different than in their zygotic counterparts in that the living maternal megagametophyte tissue surrounding the embryo is absent. The megagametophyte contains the majority of the seed storage reserves in loblolly pine and the lack of the megagametophyte tissue poses a significant challenge to somatic embryo germination and growth. We investigated the differences in seed storage reserves between loblolly pine mature zygotic embryos and somatic embryos that were capable of germination and early seedling growth. Somatic embryos utilized in this study contained significantly lower levels of triacylglycerol and higher levels of storage proteins relative to zygotic embryos. A shift in the ratio of soluble to insoluble protein present was also observed. Mature zygotic embryos had roughly a 3:2 ratio of soluble to insoluble protein whereas the somatic embryos contained over 5-fold more soluble protein compared to insoluble protein. This indicates that the somatic embryos are not only producing more protein overall, but that this protein is biased more heavily towards soluble protein, indicating possible differences in metabolic activity at the time of desiccation.     

Comparison between somatic and zygotic embryo development in Quercus robur L.

ABSTRACT

Somatic embryogenesis from juvenile explants as an efficient way for oak clonal propagation is drastically limited by the low rate of embryo germination. A comparison of the development of immature somatic and zygotic embryos, and a study of the changes in sugar content and lignin accumulation during somatic versus zygotic embryo development were conducted in view of understanding the effect of reserve substance deficiency upon somatic embryo maturation. A morphological comparison of somatic and zygotic embryos led to the identification of 4 to 7 similar developmental stages in both types of embryos, thus indicating that the accumulation phase in both zygotic and somatic embryos occurs at the same stage, when the cotyledons became thicker and opaque. Carbohydrate analysis showed the presence of glycerol, inositol, mannitol, galactose, trehalose, xylose, arabinose, glucose, fructose and sucrose in all stages of zygotic and somatic embryo development, but in different amounts. The amount of glycerol, inositol, glucose and sucrose during the early stages is larger in zygotic embryos than in somatic ones, but the time course of their accumulation is similar in both types of embryos. Lignin content, which increased continuously during development, showed a similar behaviour in zygotic and somatic embryos. In somatic embryos which were able to germinate, lignin content was higher than in nongerminating embryos at the same stage.     

Phenomenon of polyembryony. Genetic heterogeneity of seeds

Different concepts of polyembryony and genetic heterogeneity of seeds in flower plants have been reviewed. Different types, ways, and forms of plant reproduction appeared in the course of evolution as a consequences of the attached mode of life and autotrophy. This is ascribed to totipotency, “stemminess” of plant cells, and presence of constantly functioning meristems, which determined to a great extent the system of plant safety. There are two ways of formation of a new individual: sexual process → gamospermy involving meiosis and gamete fusion and asexual process → agamospermy without meiosis and gamete fusion and two types of reproduction: seed and vegetative. Both processes may take place simultaneously in one seed, as a result of which many embryos of different origins are formed: uniparental and biparental inheritance. Traditionally, this phenomenon is called polyembryony. It comprises embryoidogeny (a new category of vegetative reproduction): formation of somatic embryos (= embryoids) in the flower, seed, and on vegetative organs. Genetic heterogeneity is one of the most important characteristics of seeds, which is based on different phenomena, such as embryogeny, embryoidogeny, and gametophytic and sporophytic apomixis. When describing two types of polyembryony, sporophytic (nucellar, integumental, cleavage) and gametophytic (synergidal, antipodal), a great attention is paid to characterization of initial cells of the sexual and adventive embryos. A new concept of apogamety is developed from new positions (totipotency and “stemminess”), which is based on different genesis of cells of the egg and antipodal systems. Five possible pathways of formation of the adventive embryos have been proposed from cells of the egg apparatus. Specific features of the formation of adventive embryos in the case of gametophytic apomixis, such as androgenesis and semigamy, are discussed. Morphogenesis of the sexual and adventive embryos proceeds in the mother organism and is determined by the origin and formation of their initials, types of ovule and embryo sac, and specific features of developmental biology. This determines parallelism in their development. The main difference consists in the way of reproduction: heterophasic and homophasic. The phenomenon of polyembryony and genetic heterogeneity of seeds is essential for development of the theory of reproduction and applied research related to seed productivity of plants.     

ADVENTIVE EMBRYOGENESIS IN CITRUS AND ITS RELATION TO POLLINATION AND FERTILIZATION

Cytological and histological studies of seeds from three facultative apomictic Citrus cultivars show that adventive embryos develop, as a rule, from the first few cell layers of the nucellus adjacent to the embryo sac in the micropylar half and occasionally from the chalazal end. The adventive embryos initiated in nucellar tissue away from the embryo sac and most of those initiated from the chalazal end of the nucellus do not develop beyond the one-celled stage. When two or more embryos are developing in the same seed, the successful development of a given embryo depends on its location in relation to access to nutrients from the endosperm. The presence of a zygote and triploid endosperm in seeds with adventive embryos, the abortion of seed when endosperm degenerates, and the lack of seed set without pollination indicate that pollination and fertilization are essential for in vivo adventive embryogenesis.     

Embryological Observation on the Apomictic Rice-ApⅢ (Shuang 13)

Since 1988s, the phenomenon of multiple seedlings and multiple embryoes of apomictic rice, Ap Ⅲ (Shuang 13) has been interestingly concerned. Embryological investigation of Ap Ⅲ has been carried out with the whole dissection and conventional sectioning technique. The main conclusions are as follows: 1.8. 9 % of 5000 matured caryopsises, contained one embryo in a caryopsis, from which single seedling arose, while 8.9 % twin embryoes and 1.2 % triplet embryoes of which developed two seedlings and triple seedlings respectively; 2. In over 700 young caryopsises, 90.0%～95.0% contained only one embryo, 5.0%～7.0% twin embryoes, except a few of the ovules with absorptive embryo sac or embryo and endosperm. Triplet embryoes caryopsis could not be traced further due to their low frequency; 3. Authors' materials showed that all the single, twin and triplet embryoes were situated at the micropylar end in one embryo sac of young ovule and no adventitious embryo was seen outside the embryo sac of ovules. From the aforementioned results, it can be infered that albeit single embryo is derived from the egg cell yet it could be possible twin or triplet embryocs may be developed from the egg cell and one or two of the synergids either or not through fertilization.     

黑穗醋栗(Ribes nigrum L.)未受精胚珠培养诱导体细胞胚状体和植株再生的研究

本文报告了黑穗醋栗(Ribes nigrum L.)三个栽培品种中的薄皮黑豆未受精胚珠(花粉发育到单核晚期)在MS基本培养基附加植物激素BA,2.4-D和GA_3中形成了体细胞胚状体。长度约为0.5—1.0cm大小的胚状体在生根培养基中可以形成完整的再生植株。经过筛选得到了体细胞胚性愈伤组织无性系,继代培养二年多仍能保持胚状体形成能力。试验结果表明:诱导体细胞胚胎发生受品种和接种时期的影响,适宜的接种时期为花粉发育到单核晚期。培养基中的BA为诱导胚胎发生和保持胚性愈伤组织无性系所必需。     

檀香种胚离体培养快速育苗研究

利用添加植物生长调节剂的培养基对檀香种胚进行离体培养,研究种胚萌发和成苗的影响因素以及种苗快速繁育技术。结果显示:生长调节剂GA₃和种胚成熟度都是影响檀香种胚萌发和成苗的重要因素,檀香成熟胚在含14 μmol/L GA₃的MS培养基上7~10 d即可萌发,45 d后成苗率达82.5%,檀香种胚离体培养是有效的快速育苗方法。