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

The transition from free nuclear to cellular endosperm of Coix lacryma-jobi was eompleted 2 days after pollination. By 3 days after pollination the central cell was filled with endosperm cells. At first all cells of endosperm underwent division, later cell division was limited mainly in the peripheral region. 10 days after pollination the epidermal layer ceased its periclinal division and became the aleurone layer. Cell division persisted in the subepidermal 'cambium-like layers until the caryopsis nearly matured. Ceils of the inner region of endosperm became enlarged. Several layers of transfer cells were formed at the basal part of the endosperm. Starch grains appeared in endosperm cells on the 9th day after pollination. 10 days after pollination, lipid bodies occurred in the aleurone layer and the underlying layers. 13 and 15 days after pollination, the small vacuoles of aleurone cells contained protein and 20 days after pollenation they became aleurone grains. By 15 days after pollination pro tein bodies were formed in starch endosperm. Storage reserve deposition continued until the grain ripened. A correlation between endosperm and emoryo development was also observed.     

薏苡种子胚芽鞘细胞的结构

观察了薏苡浸泡种子胚芽鞘的结构。胚芽 外,内表皮薄壁组织及２个侧位的维管束组成。在外表皮两处,观察到径向壁不边原细胞群,它们实际是合胞体。薄壁细胞含丰富的核糖体,内质网小泡和线粒体,说明代谢活动已经活跃。初生纹孔场内有胞间连丝,显示胞间已存在物质的共质运转。     

黔西南薏苡内生真菌多样性及其抗菌活性研究

为进一步了解贵州黔西南薏苡(Coix lacryma-Jobi)可培养内生真菌的多样性组成及其抗菌活性。通过组织块分离法,选取薏苡根、叶和种仁为分离对象,进行内生真菌分离,并通过形态学观察,分子生物学特征对所分离内生真菌进行鉴定。采用平板菌块对峙法对薏苡内生真菌进行抗菌活性筛选。从薏苡根、叶和种仁中共分离纯化获得76株薏苡内生真菌,根据形态和分子生物学特征,它们属于10个目、14个科、26个属和1个不明属以及未知菌;以大肠杆菌(Escherichia coli)、金黄色葡萄球菌(Staphyloccocus aureus)、枯草芽胞杆菌(Bacillus subtilis)、酿酒酵母(Saccharomyces cerevisiae)为指示菌株,对薏苡内生真菌进行抗菌活性筛选。其中有25株对大肠杆菌具有抗菌活性,占分离菌株的比例为32. 9%;有34株对金黄色葡萄球菌具有抗菌活性,其比例为44. 7%;有24株对枯草芽胞杆菌具有抗菌活性,比例是31. 6%;有23株对酿酒酵母具有抗菌活性,占比例为30. 3%;对4种标准菌株都有抗菌活性的有13株,占总株数的17. 1%。薏苡的不同组织结构中存在丰富的内生真菌资源,部分内生真菌具有抑制其他微生物生长的活性,具有产生天然活性产物成分的潜力,为工业应用奠定了资源基础,具有进一步发掘和研究的价值。     

水稻程序性细胞死亡抑制基因Dad-1在薏苡中的定位

基因Dad-1是普遍存在于动物和植物中一个高度保守的程序性细胞死亡抑制基因。以水稻Dad-1基因为探针,采用Southern杂交在薏苡中检出了Dad-1基因的同源序列,并利用荧光原位杂交的方法对其进行了染色体物理定位。在薏苡第9染色体短臂上检测到Dad-1基因的杂交信号,信号与着丝粒的百分距离为67.44±1.45。     

通过图像分析方法作出的薏苡定量染色体图

薏苡染色体数目为2n=20,由于中期染色体的长度差异不大,相邻染色体难以区分,而薏苡前中期的染色体相对较长,经DAPI染料染色后染色体上显示明显不同的深染区和浅染区,深染区和浅染区分别相当于染色体上染色质纤维较浓缩和较伸展的区域。前中期染色体上染色深浅不一的染色区可以作为识别薏苡特定染色体的重要标志,也可用于辨别同源染色体。用MetaMorph软件定量分析了薏苡每条前中期染色体上(从短臂到长臂)DAPl信号强度的变化,结合染色体的长度和臂比作为辅助参数,构建了薏苡前中期染色体的定量染色体图。该定量染色体图不仅描述了每条染色体不同区域浓缩程度的不同,而且也反应出不同浓缩区域所占染色体长度的比例。因此该定量染色体图是实际的前中期染色体的一种直观模式,可作为识别薏苡基因组中每条染色体的有力依据。     

基于SRAP标记的薏苡种质资源遗传多样性及DNA指纹图谱构建

利用SRAP分子标记对从各主要产地收集到的90份薏苡种质进行遗传多样性分析,其中68份收集于福建省,6份来自中国台湾,16份来自浙江、辽宁、山东、河南、云南、江苏、湖南、广东、上海等省(市)。结果表明,从88对SRAP引物组合中筛选出26对引物进行SRAP扩增,共扩增出185条带,其中具有多态性的有157,占总数的84.86%,表明90份薏苡种质表现出丰富的遗传多样性。基于SRAP标记利用系统聚类法将90份薏苡种质资源分为4大类,与形态性状分类结果有一定的相似性;利用16对SRAP引物构建了73份薏苡种质资源的DNA指纹图谱,为薏苡遗传研究、品种选育与资源保护提供了依据。     

Microsporogenesis,Megasporogenesis and the Formation of Male and Female Gametophyte in Coix lacryma-jobi L.

The development of the anther wall follows the monocotyledonous type. During meiotic stages of prophase I, some cytoplasmic channels are observed on the walls between meiotic cells. which divide synchronously. Cytokinesis in the microspore mother cell is of the successive type and gives rise to iscbilateral tetrad. The cell wall between the generative cell and the vegetative cell in early stage shows PAS positive reaction. The mature pollen grain is of 3-celled type. The development of the female gametophyte follows the polygonum-type; the antipodal cells proliferate to form a multicellular tissue mass. Many starch grains are present in the central cell. The nucleus of the mature egg cell is located at the micropylar end; a great deal of starch grains are in the cytoplasm surrounding the nucleus, while vacuoles of various size distribute throughout its cytoplasm but are more and larger at the chalaza,1 end. The nucleus of the synergid cell is located at the micropytar end where a filiform apparatus is formed and many small vacuoles are present at the chalazal part.     

薏苡种子形态性状多样性评价

薏苡是优质的药食兼用作物。本研究对我国36份薏苡种质的种子形态性状进行了主成分分析和聚类分析。结果表明,36份薏苡种质具有较高的形态性状多样性。不同薏苡种质的4个种子质量性状间存在差异。单因素方差分析显示11个种子数量性状在不同薏苡种质间呈极显著差异。数量性状的变异系数在8.43%~68.01%之间,种壳与外种皮重量的变异系数最大而种仁长度的变异系数最小。主成分分析将36份薏苡种质分为中部及南部栽培种质、北部栽培种质和野生种质3大类,3类种质的形态特征存在明显差异。聚类分析将36份薏苡种质分为5大类群,即中部及南部栽培种质、北部栽培种质和3个类群野生种质。初步筛选出6份可能具有较高抗肿瘤活性的薏苡种质FJNJ、TJ-BK-1、TJ-BK-2、MY-BK-1、MY-BK-2和JYSD。     

薏苡胚乳传递细胞的超微结构

传粉后１０ ｄ,薏苡（Ｃｏｉｘ ｌａｃｒｙｍ ａ－ｊｏｂｉＬ．）颖果基部胚乳最外层传递细胞具长而多的壁内突,第二层细胞的壁内突较第一层的短而少,均具瓣裂的细胞核、丰富的线粒体、粗糙内质网、核糖体、产生小泡的高尔基体及与壁内突质膜相连的、含深色物质的囊泡。线粒体分布于壁内突附近或其间。授粉后２５ ｄ,第一、二层细胞壁内突发达,几乎充满了细胞,但细胞器可见。第四层传递细胞具树枝状及网状的壁内突,大量线粒体、具质体膜的淀粉粒、脂体存在壁内突附近或壁内突的间隙内。高尔基体常见,仅见很少的片段内质网。第五层传递细胞具短的壁内突、较大的淀粉粒及许多小蛋白质体。两个时期的第一、二层细胞内均未观察到胞间连丝。授粉后２５ ｄ,第四层及以上的传递细胞的细胞壁和呈网状的壁内突均含有胞间连丝。还讨论了各种细胞器的作用及各层传递细胞的功能     

玉米mir1基因在玉米和薏苡中的比较物理定位

玉米基因mir1编码一种抗秋季黏虫的半胱氨酸蛋白酶。利用RFLP作图mir1基因被定位在玉米第 6号染色体短臂上 ,但它在第 6号染色体短臂上的物理位置还不知道。实验以mir1和 4 5SrDNA为探针 ,通过双色荧光原位杂交技术确定了mir1基因在玉米细胞分裂中期和粗线期第 6号染色体上的物理位置。Southern杂交结果表明 ,在薏苡基因组中存在mir1基因的同源序列 ,进一步利用荧光原位杂交的方法确定mir1基因的同源序列定位于薏苡第 7号染色体长臂的近末端 ,其信号与着丝粒的百分距离为 73 33± 0 15。     

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.     

Somatic embryogenesis and plant regeneration from immature inflorescence segments of Coix lacryma-jobi

Callus was obtained from segments of immature inflorescence of Coix lacryma-jobi cultured on N6 medium containing 1–2 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 3–5% sucrose. Plantlets were regenerated when embryogenic calluses were transferred onto MS medium with 0.5 mg/l kinetin and 0.01 mg/l naphthaleneacetic acid (NAA). Regenerated plants had the diploid chromosome number (2n=20).     

Induction of Endosperm Calluses and Regeneration of Endosperm Plantlets of Asparagus officinalis

The endosperm callus has been induced from the young endosperm of Asparagus officinalis L. on the MS supplemented with auxin. The induction frequency of callus amounts to 65.9%–83.1%. When the callus was transferred to the medium supplemented with lower concentration of NAA 0.1 ppm or containing BA 1 ppm and NAA 0.5 ppm, the differentiation of shoots, roots and a few embryoids began to occur. A few calluses and embryoids can develop into plantlets. The chromosome number in the cells from the same root tip and shoot apex of endosperm plantlet is very unstable. They can be euploids (n=10, 2n=20, 3n=30, 4n=40). or aneupl0ids (n=6, 7, 17, 25, 53).     

苏铁白化苗叶片细胞的超微结构

本文系苏铁白化苗研究系列报道之二。应用超薄切片和电镜观察技术对苏铁白化苗叶片细胞的超微结构观察,结果表明,苏铁白化苗叶片细胞的超微结构与绿苗的相比有明显不同,前者的细胞质含量稀少,未见有结构完整的叶绿体和线粒体,但见有呈支解、游离状的类似叶绿体片层的膜结构和内嵴几乎完全消失的退化线粒体存在;后者的细胞结构正常。     

五种豆科种子胚乳细胞的微观结构的比较

运用光学显微镜和电子显微镜分别对胡芦巴、野皂荚、皂荚、瓜尔豆、塔拉豆等五种豆科种子内胚乳细胞的形态、大小、壁厚等微观结构进行了观察比较。研究结果表明：五种种子内胚乳细胞壁几乎完全由胶状的透明的半乳甘露聚糖所填充,细胞的形态、大小差异较大,瓜尔豆、塔拉豆的内胚乳细胞呈球形或近球形;胡芦巴内胚乳细胞为多角形或多角椭圆形,它们细胞壁界限清晰,细胞排列整齐有规律;皂荚、野皂荚则为不规则的长椭球形,细胞壁彼此连接成片,界限模糊。一般地说：当细胞壁壁厚小于4μm时,胚乳胶粉中的水不溶物含量较低,1％浓度的胶液的粘度较高,而当壁厚大于4μm时,情况反之。本文的研究目的是为植物胶生产、加工提供一定的细胞学方面的依据。     

Ultrastructure of the Malpighian Tubule Cells in the Mosquito Larvae, Anopheles sinesis

Ultrastructure of epithelial cells constituting the Malpighian tubule of Anopheles sinesis last instar larvae was observed with electron microscope. Malpighian tubule consists of four long and narrow tubule structures with principal cells in typical absorptive cells and regenerative cells forming the simple epithelium. Apical plasma membrane of the principal cell is differentiated into microvilli with one mitochondrion in each microvilli. Basal plasma membrane had extreme infolding to form a canaliculi and a well developed mitochondria was attached in the infoldings. And, rER, ribosomes, and vacuoles were well developed inside the cells. However, there were two main cell types depending on the differentiation of cell organelles. Type 1 cell was cubic, forming the distal portion of Malpighian tubule. The length of microvilli was approximately 4 μm and the basal infoldings were introjected to the depth of 2 μm inside the cell. On the other hand, Type II cell that formed the main proxinal portion was a low squamous type cells with shorter 2 μm of microvilli and the basal infoldings were introjected to the depths of 4 μm inside the cell. As for vacuoles scattered inside the cells, they were regularly observed in both Type I and II and the Type II cells had better developed cellular organelles. Although regenerative cells were extremely small, their cellular organelles were developed and their overall electron density was high that they appeared darker than the principal cells.     

薏苡生药学研究进展

薏苡(Coix lacryma-Jobi L.var.ma-yuen (Roman.) Stapf)为禾本科蜀黍族薏苡属植物,其果仁薏苡仁是我国传统中药材之一,具有利水渗湿,健脾止泻,清热排脓之功效.目前对薏苡的研究主要集中在药理活性和功能食品开发方面,缺乏对其生药学的系统研究.因此长期以来对薏苡的研究存在着起源不清、品种杂乱、种质资源利用欠佳等问题.近年来随着分子生物学的蓬勃发展,多种现代生物技术已用于薏苡种质资源鉴定和遗传多样性的研究,为薏苡资源的保护和可持续发展提供了支持.本文就薏苡的起源与进化,种属分类,细胞学和分子生物学研究进展加以综述,从分子水平揭示薏苡遗传结构及与近缘植物的亲缘关系,为明确薏苡今后种质资源的开发和优异基因的利用提供理论依据.     

Ultrastructure of the Rectum Epithelial Cells in the Mosquito Larvae, Anopheles sinensis Wiedemann

ABSTRACT The Ultrastructure of rectum epithelial cells in the mosquito larvae, Anopheles sinensis Wiedemann, was studied using electron microscope. The rectal epithelium forms rectal papillae composed of the absorptive cells and the surrounding basal cells. Moreover, rectal epithelium was covered with thin cuticular intima. Apical plasma membrane of the epithelial cells had infoldings and in between them, mitochondria developed into elongated shape were attached. In addition, the membrane infoldings reach down into the cell cytoplasm to form several layers of leaflet-like prolongations. On both sides of these prolongations were also large, well-developed mitochondria. Their formation was that mitochondria were attached to 3 μm length and 4–13 layers of membrane wrinkle lump. Many spherites, which are lamelated crystals that form an illusory structure in concentric circles inside of the cytoplasm of epithelial cell were observed. Basal plasma membrane in the epithelial cells was also wrinkled to promulgate into the cytoplasm to become basal infoldings producing canaliculi in basal labyrinth formation. There were many mitochondria scattered in these formations as well. On the bottom of the epithelial cell, basal lamina was attached and between basal lamina and muscle bundle was subepithelial space, which is connective tissue. Inside the space, tracheal and nerve cells were observed.