薏苡胚发育及贮藏营养物质积累的研究

薏苡（Ｃｏｉｘ ｌａｃｒｙｍ ａ－ｊｏｂｉ）胚发育分下列各期：棒形胚前的原胚期、棒形胚期、胚芽鞘期、１叶期、２ 叶期、３叶期、４ 叶期、５ 叶期及６叶期成熟胚。３ 叶期胚具１ 条不定根（种子根）,４ 叶期具２ 条,５ 叶期及成熟胚期具３ 条。不定根与胚根排成１ 纵行。营养物质最先在盾片细胞中积累。开花后９ 天的１ 叶期胚,在盾片、胚芽鞘及胚轴细胞中积累了淀粉,以后遍及成熟胚的各部分。淀粉粒含量与器官发生及生长顺序成正相关,但发育后期,盾片细胞内的淀粉粒含量下降。开花后１０ 天,盾片细胞中形成含晶体的蛋白质体,晶体含蛋白质及植酸钙镁。以后,这种蛋白质体增多、增大。同时,又形成不含晶体的蛋白质体。一定时期,含晶体的蛋白质体消失,不含晶体的蛋白质体增多,直到胚成熟。开花后１３ 天,胚芽鞘上部细胞形成蛋白质体。以后遍及成熟胚的各部分,器官发生越早,所含蛋白质体越多、越大。开花后１０ 天,盾片细胞中产生了脂体,成熟胚的盾片细胞,含有大量的脂体。还观察了胚发育各期与颖果及盾片长度的对应关系     

大麦胚和胚乳发育的相关性及贮藏营养物质的积累

大麦(Hordeum vulgare L.)开花后1d,见合子及退化助细胞,游离核胚乳尚未形成;开花后2～3d,胚为5及10个细胞,胚乳为游离核期;开花后4及5、6d,胚为梨形及长梨形,胚乳达细胞化期;开花后8d,胚为胚芽鞘期,糊粉层原始细胞产生;开花后10d,胚具1叶,糊粉层1～2层;开花后13d胚为2叶胚,亚糊粉层发生;开花后17d,3叶胚形成,糊粉层多为3层并停止分裂,菱柱形及不规则胚乳细胞分化;开花后21～29d,胚为4叶胚,胚乳进一步分化;开花后33d,胚为5叶成熟胚,胚乳亦成熟。淀粉、蛋白质在胚中积累始于开花后13d。在盾片中由基向顶发生,在胚芽鞘及叶原基中,首先在顶端出现。成熟盾片顶端的淀粉消失。开花后6d,胚乳开始积累淀粉;开花后10d,糊粉层及胚乳细胞积累蛋白质。开花17d后胚乳的蛋白质体多聚集,29d后蛋白质体显著减少。开花后17d,在盾片及糊粉层细胞中检测到油脂。果长或果长与稃片长之比和盾片长可作为不同发育期胚和胚乳的形态指标。     

槐种子发育过程中子叶细胞显微结构的变化和过氧化物酶同工酶变化的动态

槐（SophorajaponicaL．）种子发育过程中,子叶细胞大量合成和积累储藏物质,光镜和电镜下用组织化学方法观察到开花后30d,叶肉细胞高度液泡化,其中分布着大量的质体、粗面内质网、一些线粒体和小脂体。淀粉粒开始积累。开花后60d,淀粉粒和脂体不断长大,数量明显增多,液泡中蛋白体开始形成。从花后90d开始,液泡中有大量蛋白体形成且不断长大,有些蛋白体变得不规则形。至种子成熟期,叶肉细胞几乎完全被蛋白体及脂体所填充。淀粉粒及大多数细胞器消失。基于上述观察,还对种子发育过程中子叶细胞过氧化物酶活力及同工酶港的变化进行了观察和分析。     

玉竹雄配子的发育——着重阐明质体在生殖细胞和营养细胞中的分布和变化

玉竹(Polygonatum simizui Kitag)小孢子在分裂前,质体极性分布导致分裂后形成的生殖细胞不含质体,而营养细胞包含了小孢子中全部的质体。生殖细胞发育至成熟花粉时期,及在花粉管中分裂形成的两个精细胞中始终不含质体。虽然生殖细胞和精细胞中都存在线粒体,但细胞质中无DNA类核。玉竹雄性质体的遗传为单亲母本型。在雄配子体发育过程中,营养细胞中的质体发生明显的变化。在早期的营养细胞质中,造粉质体增殖和活跃地合成淀粉。后期,脂体增加而造粉质体消失。接近成熟时花粉富含油滴。对百合科的不同属植物质体被排除的机理及花粉中贮藏的淀粉与脂体的转变进行了讨论。     

高等植物胚胎的发育生物学研究——Ⅰ.水稻、小麦胚胎发育的生长模式

本文结合稻胚的发育模式描述了水稻开花后1至30天期间胚分化发育过程,并指出了胚分化早期胚柄部分与珠被等组织的密切关系。对水稻开花后6～30天及小麦开花后8～27天期间胚鲜重、干重、体积和细胞数的发育模式进行了测定。在此基础上计算了平均每个细胞鲜重和体积的变化,以及麦胚细胞鲜重、干重和体积的变化。此外还测定了水稻胚长度,胚乳鲜重和体积的变化。结果表明;稻胚体积、鲜重在开花后6～11天,干重在6～13天期间呈指数增长,小麦胚鲜重、干重和体积变化在开花后8～15天期间指数增长趋势亦明显,稻胚每个细胞的平均鲜重和体积在开花后7天以前较小,9天以后明显增大;小麦胚每个细胞的平均鲜重、干重、和体积在开花后10天以前较小,12天以后则明显增大。稻胚在开花后1～30天期间长度变化呈S形曲线。     

西瓜种子发育和萌发过程中子叶细胞超微结构的变化

西瓜种子子叶内贮存物质开始积累时,细胞质内有大量核糖体、质体、线粒体,内质网片段和囊泡,种子脱水期至成熟期,细胞器的数量减少,成熟种子子叶细胞的细胞壁不连续,几乎观察不到细胞器的存在,种子萌发过程中内质网,线粒体,质体的数目逐渐增多,叶肉细胞的质体发育成叶绿体,种子形成过程中,在子叶细胞大液泡分隔的同时,膨胀的内质网囊泡内积累蛋白质（直径0.1-0.4μm),这些小的蛋白质球体最终进入液泡形成大的蛋白体（直径1-3μm);萌发种子贮存蛋白质被水解的同时,一些脂体进入液泡并被分解,同时液泡融合;脂类物质开始积累的时间早于蛋白质,积累的量较蛋白质多,但在萌发种子中被彻底水解的时间晚于蛋白质,淀粉粒的数量在种子形成时减少,种子萌发时在表皮细胞和叶肉细胞内都重新合成。     

长豇豆胚和胚乳的发育及营养物质积累

长豇豆(Vigna sesquipedalis (L.)Fruwirth)开花前7—10小时传粉,开花后8—10小时完成双受精。合子期珠孔端及合点部位胚囊的周界壁有壁内突。胚发育属柳叶菜型。胚柄的基部细胞及基部区域外层细胞的外切向壁发生壁内突。成熟胚中胚柄宿存。开花后9—16天为子叶细胞中淀粉积累期,开花后12—18天为蛋白质积累期。胚乳发育为核型,珠孔端胚乳细胞化,合点端保持游离核状态。胚乳外层细胞为传递型细胞,珠孔端的胚乳细胞形成折叠细胞群,亦有壁内突。心形胚期胚乳开始退化解体,成熟胚期胚乳完全消失。     

水稻胚胎形成期间蛋白质和RNA合成活力的变化

用~3H-亮氨酸和~3H-尿苷标记不同发育时期的稻胚,发现蛋白质合成活力呈四阶段变化;各种RNA的合成与胚胎各发育阶段密切有关。α-鹅膏蕈碱(1.0μg/ml)对稻胚RNA合成的抑制作用在开花后7、15和18天较强,13天时很弱。在水稻胚胎形成期间,胚细胞蛋白质合成活力高峰先后出现于胚分化后期(开花后11天)和成熟中期(18天);mRNA的合成在分化初期(7天)和成熟中期(15～18天)较强;而rRNA和/或tRNA的合成高峰则出现在胚胎器官原基分化已经完成时(13天)。     

油松细胞质遗传的细胞学机理:Ⅱ.雄性和雌性细胞器在受精和原胚时期的传递(英文)

在电镜下观察油松 (PinustabulaeformisCarr.)传粉后的胚珠临近受精时的花粉管和卵细胞的细胞质、受精时雄配子体细胞质的传递、游离核和细胞原胚发育时期质体和线粒体的传递。在成熟卵细胞中含许多线粒体 ,缺少正常结构的质体 ,它们转变为大内含体。此外 ,卵细胞还有丰富的小内含体和其他一些细胞器。花粉管在卵细胞的珠孔端释放其内含物。精核与卵核融合时 ,核周围未见来自精细胞的质体和线粒体。不参与融合的精核停留在接受液泡旁 ,在其周围有大量的雄性细胞质 ,其中混合有精细胞、管细胞和卵细胞的细胞器。在游离核原胚时期 ,核周区的细胞质中可见雄性与雌性亲本的细胞器相混合 ;其中许多线粒体与原来卵细胞中的线粒体有相同的形态 ,也有一些线粒体看来是来自精细胞和管细胞 ;质体是由雄配子体传递 ,形态与精细胞的或花粉管中的质体相似。卵细胞中变异的质体 (即大内含体 )在原胚发育时期变为液泡状 ,而雄性质体参加到新细胞质中。在原胚细胞中 ,线粒体大多数为母本来源 ,质体则表现为精细胞或管细胞的质体形态。该研究确定了油松具父系质体和双亲线粒体遗传的细胞学基础。对裸子植物线粒体和质体遗传的机理从细胞学的角度进行了分析。     

水稻种子发育过程中Poly(A)RNA和蛋白质水平的变化

我们用[~3H]—Poly(U)饱和杂交的方法分析了水稻种子发育过程中Poly(A)含量和Poly(A)RNA水平的变化。胚乳发育过程中,Poly(A)含量和Poly(A)RNA水平均于开花后11天达到高峰,比蛋白质高峰出现时间约早10天。随着胚乳的成熟,蛋白质水平在开花后6～21天持续增长。但 Poly(A)含量和Poly(A)RNA水平却急剧下降。因此,在胚乳发育早期合成的Poly(A)RNA中,可能有部分不是直接用于蛋白质的合成。在胚的发育过程中,Poly(A)含量和Poly(A)RNA水平分别出现三次高峰。开花后30天,每胚含有5.94ng Poly(A)RNA,约占胚总RNA的0.097％,为稻胚中贮存的mRNA存在提供了一个直接的证据。     

Development of the cotyledon cells during olive (<Emphasis Type="Italic">Olea europaea</Emphasis> L.) in vitro seed germination and seedling growth

The structural changes occurred in differentiating olive cotyledon cells into mesophyll cells are described. Using histological and immunocytological methods as well as microscopic observations, we showed that in the cells of mature embryo, large electron-dense proteins bodies (PBs) are surrounded by numerous oil bodies (OBs). After 3 days of in vitro germination, the presence of large PBs originated by fusion of smaller PBs was observed. It was also detected a close spatial proximity between PBs and OBs, likely as a reflection of interconnected metabolic pathways. Between the 3rd and the 12th day of germination, the formation of a large vacuolar compartment takes place accompanied by a decrease in the PBs and OBs number. This was coincident with a progressive decrease in the amount of the 11S-type seed storage proteins (SSPs), showed in situ and after Western blot analysis of crude protein extracts. After 26 days germination, the cellular organization became typical for a leaf mesophyll cell, with well-differentiated chloroplasts surrounding a large central vacuole. Our results suggest that the olive cotyledon storage reserves are mobilized gradually until the seedling becomes autotrophic. Moreover, the specific accumulation of storage proteins in the intravacuolar material suggests that these structures may operate as a shuttle for SSPs and/or products of their degradation into the cytoplasm, where finally they supply amino acids for the differentiating mesophyll cells.     

Studies on Embryo Development and Deposition of Storage Reserves in Coix lacryma-jobi

Embryo development in Coix lacryma-jobi is classified into the following stages: proembryo before club-shaped, club-shaped, coleoptilar, I-leafed, 2-1eared, 3-1eared, 4-1eared, 5-leafed and 6-leafed (mature embryo). The 3-, 4-, 5-leafed embryos have 1, 2 and 3 adventitious roots (seminal roots) respectively, and the matrue also has 3. These seminal roots are arranged in a longitudinal row parallelling with the radicle. The storage reserves first deposit in the scutellar cells. 9 days after anthesis (l-leafed stage), the starch grains are accumulated in cells of scutellum, coleoptile and mesocotyle. When the embryo matures, starch grains are deposited throughout its cells. The increase in size and amount of starch grains correlates with the initiation and growth order of the embryonic organs. But the amount in the scutellar cells decreases from later to mature stage. 10 days after anthesis (2-leafed stage), protein bodies containing crystals, of protein and phytin are present in the scutellar cells. They subsequently become larger and abundant druses. At the same time some protein bodies without crystals are also formed. Later, the protein bodies containing crystals disappear, while those without crystals increase until the embryo matures. 13 days after anthesis (3- leafed stage) protein bodlies are formed in the upper coleoptile cells. Protein bodies are rich in the cells of mature embryo, but the earlier the organ of embryo occurs, the more and the larger protein bodies it contains. 10 days after anthesis, lipid bodies appear in the scutellar cells and increase in size and quantity rapidly as the embryo develops. The correlation of the length of caryopsis and scutellum with embryo development is also observed.     

Arabidopsis β-ketoacyl-[acyl carrier protein] synthase i is crucial for fatty acid synthesis and plays a role in chloroplast division and embryo development

Lipid metabolism plays a pivotal role in cell structure and in multiple plant developmental processes. β-Ketoacyl-[acyl carrier protein] synthase I (KASI) catalyzes the elongation of de novo fatty acid (FA) synthesis. Here, we report the functional characterization of KASI in the regulation of chloroplast division and embryo development. Phenotypic observation of an Arabidopsis thaliana T-DNA insertion mutant, kasI, revealed multiple morphological defects, including chlorotic (in netted patches) and curly leaves, reduced fertility, and semidwarfism. There are only one to five enlarged chloroplasts in the mesophyll cells of chlorotic sectors of young kasI rosette leaves, indicating suppressed chloroplast division under KASI deficiency. KASI deficiency results in a significant change in the polar lipid composition, which causes the suppressed expression of FtsZ and Min system genes, disordered Z-ring placement in the oversized chloroplast, and inhibited polymerization of FtsZ protein at mid-site of the chloroplast in kasI. In addition, KASI deficiency results in disrupted embryo development before the globular stage and dramatically reduces FA levels (~33.6% of the wild type) in seeds. These results demonstrate that de novo FA synthesis is crucial and has pleiotropic effects on plant growth. The polar lipid supply is important for chloroplast division and development, revealing a key function of FA synthesis in plastid development.     

Membrane-bound Plastid Inclusions and Chloroplast Thylakoid Formation in Sunflower (Helianthus annuus L.)

The ultrastructure, distribution and frequency of membrane-boundplastid inclusions present in the epidermal cells of leavesof intact sunflower plants (Helianthus annuus L.) and in theepidermal and mesophyll cells of sunflower leaf discs culturedin darkness have been studied. These inclusions appear to bedilated thylakoids containing a granular material which, undernormal conditions, is probably involved in chloroplast membraneformation. It is suggested that this material accumulates, andinclusions form, in the chloroplasts of sunflower leaves intwo specific situations. Firstly, in the completely differentiatedcells of the epidermis where the chloroplasts, although at arelatively immature stage, have nevertheless reached a terminalstage of development. Secondly, in the mesophyll cells of youngleaves when chloroplast development has been arrested at animmature stage by a 5-day dark period. In the latter situationthe material can be remobilized if plastid development is restimulated.The plastids of sunflower leaf discs cultured in darkness containboth membrane-bound inclusions and prolamellar bodies, indicatingthat they are separate and distinct structures possibly containingdifferent membrane components. Helianthus annuus L., sunflower, chloroplast, ultrastructure, plastid inclusions, thylakoid formation     

Oil bodies in leaf mesophyll cells of angiosperms: overview and a selected survey

Neutral (storage) oil bodies occur in leaf mesophyll cells of many angiosperms, but their literature has been largely forgotten. We review this literature and provide a survey of 302 species and hybrids from mostly north-central US species representing 113 families. Freehand cross sections of fresh leaves stained with Sudan IV verified the presence of oil. In 71 species from 24 families we observed 1-15 oil bodies per mesophyll cell. The eudicot families Asteraceae, Caprifoliaceae, Lamiaceae, and Rosaceae had the highest number of species with oil bodies, whereas few or no species in the Apiaceae, Betulaceae, Fabaceae, and Scrophulariaceae had them. Only three of 19 monocot species sampled had oil bodies. Repeat sampling of a Malus (crabapple) cultivar and a Euonymus species showed conspicuous oil bodies in mid-summer and also in mid-autumn in both attached and recently shed leaves. Oil bodies in leaf mesophyll cells are conspicuous (visible in hand cross sections using moderate magnification in unstained water mounts) in numerous species, and they occur throughout the growing season in at least some species. Neutral oil bodies in leaf mesophyll cells are not mentioned in contemporary textbooks and advanced works, but they deserve recognition as significant cellular components of many taxa, in which they may be significant sources of commercial oils.     

Plastidic protein Cdf1 is essential in Arabidopsis embryogenesis

Arabidopsis cell growth defect factor-1 (Cdf1 in yeast, At5g23040) was originally isolated as a cell growth suppressor of yeast from genetic screening. To investigate the in vivo role of Cdf1 in plants, a T-DNA insertion line was analyzed. A homozygous T-DNA insertion mutant (cdf1/cdf1) was embryo lethal and showed arrested embryogenesis at the globular stage. The Cdf1 protein, when fused with green fluorescent protein, was localized to the plastid in stomatal guard cells and mesophyll cells. A promoter-β-glucuronidase assay found expression of Cdf1 in the early heart stage of embryogenesis, suggesting that Cdf1 was essential for Arabidopsis embryogenesis during the transition of the embryo from the globular to heart stage.     

The Rest Period of Rhododendron Flower Buds: III. CYTOLOGICAL STUDIES ON THE ACCUMULATION AND BREAKDOWN OF PROTEIN BODIES AND AMYLOPLASTS DURING FLOWER DEVELOPMENT

Rhododendron flower development occurs in three easily definedstages: a pre-rest stage, during which petal growth is mainlyby cell elongation; an indeterminate rest period; and an after-reststage, that begins when the flowers resume growth and ends atanthesis. Early in the pre-rest stage of development, protein bodies andamyloplasts accumulate in the petals. The epidermal cells accumulateonly protein bodies and the mesophyll cells accumulate amyloplaststhat have a few small protein bodies around the periphery. Thesubepidermal cells and the cells around the vascular bundlesaccumulate both large protein bodies and amyloplasts. Duringthe rest period there is a cessation of cell elongation andthe reserve protein bodies and amyloplasts remain intact. The protein bodies in all of the cells including those aroundthe amyloplasts are proteolized early in the after-rest stageof development. Digestion of the starch granules occurs whenthe petals are about one-half their final size. Epidermal-cell expansion during after-rest is relatively uniform;the walls between adjacent epidermal cells remain attached toeach other. The mesophyll cells elongate irregularly and thewalls of adjacent cells separate giving rise to large intercellularspaces. At anthesis the petal cells consist of a cell wall, a parietalcytoplasm, and a large central vacuole.     

Fusion of oil bodies in endosperm of oat grains

Few microscopical studies have been made on lipid storage in oat grains, with variable results as to the extent of lipid accumulation in the starchy endosperm. Grains of medium- and high-lipid oat (Avena sativa L.) were studied at two developmental stages and at maturity, by light microscopy using different staining methods, and by scanning and transmission electron microscopy. Discrete oil bodies occurred in the aleurone layer, scutellum and embryo. In contrast, oil bodies in the starchy endosperm often had diffuse boundaries and fused with each other and with protein vacuoles during grain development, forming a continuous oil matrix between the protein and starch components. The different microscopical methods were confirmative to each other regarding the coalescence of oil bodies, a phenomenon probably correlated with the reduced amount of oil-body associated proteins in the endosperm. This was supported experimentally by SDS-PAGE separation of oil-body proteins and immunoblotting and immunolocalization with antibodies against a 16 kD oil-body protein. Much more oil-body proteins per amount of oil occurred in the embryo and scutellum than in the endosperm. Immunolocalization of 14 and 16 kD oil-body associated proteins on sectioned grains resulted in more heavy labeling of the embryo, scutellum and aleurone layer than the rest of the endosperm. Observations on the appearance of oil bodies at an early stage of development pertain to the prevailing hypotheses of oil-body biogenesis.     

Transmission of male cytoplasm during fertilization in Nicotiana tabacum

Serially sectioned embryo sacs of Nicotiana tabacum were examined during fertilization events using transmission electron microscopy. After pollen tube discharge, the outer membrane of the sperm pair is removed, the two sperm cells are deposited in the degenerate synergid and the sperm cells migrate to the chalazal edge of the synergid where gametic fusion occurs. During fertilization, the male cytoplasm, including heritable organelles, is transmitted into the female reproductive cells as shown by: (1) the cytoplasmic confluence of one sperm and the central cell during cellular fusion, (2) the occurrence of sperm mitochondria (distinguished by ultrastructural differences) in the zygote cytoplasm and adjacent to the sperm nucleus, (3) the presence of darkly stained aggregates which are found exclusively in mature sperm cells within the cytoplasm of both female cells soon after cell fusion, and (4) the absence of any large enucleated cytoplasmic bodies containing recognizable organelles outside the zygote or endosperm cells. The infrequent occurrence of plastids in the sperm and the transmission of sperm cytoplasm into the egg during double fertilization provide the cytological basis for occasional biparental plastid inheritance as reported previously in tobacco. Although sperm mitochondria are transmitted into the egg/zygote, their inheritance has not been detected genetically. In one abnormal embryo sac, a pair of sperm cells was released into the cytoplasm of the presumptive zygote. Although pollen tube discharge usually removes the inner pollen-tube plasma membrane containing the two sperm cells, this did not occur in this case. When sperm cells are deposited in a degenerating synergid or outside of a cell, this outer membrane is removed, as it apparently is for fertilization.