Mapping of the rice （Oryza sativa L.） thermo-sensitive genic male sterile gene tms5 with EST and SSR markers

VELOPING“TWO-LINE”HYBRID RICE.THE POLLEN FERTILITY OF TGMS IS REGULATED BY THE TEMPERATURE OF ENVIRONMENT.THE POLLENS OF TGMS LINES ARE STERILE WHEN THE ENVI-RONMENT TEMPERATURE IS ABOVE A CRITICAL POINT,BUT FERTILE BELOW THIS POINT.SO FAR,A NUMBER OF T…     

包台型水稻不育系花药发育过程中的Ca2+分布

用焦锑酸钾沉淀法研究了包台型不育水稻及其保持系花药发育早期细胞中的Ca²⁺分布变化.从花粉母细胞时期开始,不育系与保持系花药中的Ca²⁺分布出现差异,不育系药壁外层和药室内壁及药隔中均有Ca²⁺出现,而保持系花药中基本没有Ca²⁺分布,在减数分裂时期表现更加明显,不育系药壁和药隔中的Ca²⁺相对于花粉母细胞时期明显地有所增加,特别是绒毡层细胞和药隔中木质部细胞的次生加厚壁上Ca²⁺增加尤其明显,而保持系花药中的Ca²⁺较少.笔者从花药发育的更早期去寻找水稻雄性不育的机理.     

In situ localization of calmodulin mRNA in the developing tobacco anthers

The temporal and spatial expression patterns of calmodulin mRNA in the developing tobacco anthers were investigated by in situ RNA hybridization, using digoxigening-labeled anti-RNA probe. Calmodulin mRNA was distributed in various developmental stages of tobacco anthers, but the expression level had temporal and spatial differences distinctly. During early stage of anther development, the expression level of calmodulin mRNA was significantly high, mainly distributed in epidermis, tapetum and transfusion parenchyma cells and so on. Especially, more mRNA was accumulated in the nuclei and chromosomes of microspore mother cells prior to and during meiosis. With the development of anther, mRNA was decreased gradually in the anther wall and pollen. By mature pollen stage, only a stronger positive reaction still existed in the epidermis of anther wall and transfusion parenchyma cells. The results suggest that the temporal and spatial expression of calmodulin mRNA is closely correlated with cell division, pollen development and substance transport.     

水稻ⅡB型拓扑异构酶OsSpo11基因的cDNA克隆、表达谱分析及其可变剪接的检测

 Spo11基因是减数分裂双链断裂（DSBs）与分裂重组的初始化基因,在基因组进化方面具有重要作用。通过改进的Nest-PCR方法,从水稻愈伤组织中克隆到了水稻DNA拓扑异构酶OsSpo11基因的cDNA片段。克隆测序与表达研究表明,OsSpo11基因是水稻ⅡB型拓扑异构酶A亚基基因,在水稻不同组织中具有低丰度表达、组织特异性、可诱导性以及前体RNA剪接的可变性等特点。可变剪接异构体的发现,使OsSpo11基因在DNA代谢、发育、遗传重组等过程中的生物学功能表现出一定的复杂性,其功能和生物学意义有待进一步研究。     

Construction and characterization of a normalized whole-life-cycle cDNA library of rice

A cDNA library with genomic complete coverage is a powerful tool for functional genomic studies.For studying the functions of rice genes on a large scale,a normalized whole-life-cycle cDNA library is constructed based on the strategy of saturation hybridization with genomic DNA using rice cultivar Minghui 63,an elite restorer line for a number of rice hybrids that are widely cultivated in China,This library consists of cDNA from 15 directionally cloned cDNA libraries constructed with different tissues from 9 developmental stages.For normalization,the denatured plasmids purified from the 15 directionally cloned libraries are mixed and hybridized with saturated genomic DNA labeled with magnetic beads in two complementary systems. Well-matched plasmids are captured from the hybridized genomic DNA and electroporated into competent DH10B E. coli for construction of the normalized whole-life-cycle cDNA library.This library consists of 62000 clones with an average insert length about 1.4kb.Inverse Northern blotting shows that this cDNA library included many rarely expressed genes and tissue-specific genes.Sequencing of 10750 cDNA clones of this library reveals 6399 unique EST s(expressed sequence tags),indicating that the non-redundancy of the library is about 59.5%.This library has been used to make cDNA microarrays for functional genomic studies.     

用抑制差减杂交技术分离烯丙异噻唑诱导水稻特异表达的基因

烯丙异噻唑(PBZ)处理水稻根能使其产生对稻瘟病的系统获得性抗性,因此在东南亚稻区被广泛用于防治稻瘟病,然而关于其作用的分子机理还知之甚少．运用抑制差减杂交技术,试图通过分离鉴定受PBZ诱导调控的关键基因,探索其作用的分子机理．以PBZ处理后的水稻叶片cDNA为目标群体(tester),以未处理水稻叶片cDNA为对照群体(driver),用经过对照cDNA差减的、烯丙异噻唑处理的cDNA群体构建了一个含260个重组子的差减文库．通过差示筛选鉴定出了26个。PBZ诱导水稻特异表达和增强表达的候选克隆．对26个cDNA克隆进行了双向测序和同源性比较,发现其中3个克隆：rJAB1,rTAB2和蛋白磷酸酯酶2Aδ调节亚基同型物基因,位于抗病相关信号转导途径上,它们与哺乳动物和人类免疫途径上的信号因子有明显相似之处,因此推断可能与诱导抗性有关．另外8个克隆与已知基因同源性为70％～99％．经Northern杂交分析,其中rJAB1(编码c-jun激活区结合蛋白1)受烯丙异噻唑和稻瘟菌诱导表达;膜糖蛋白同源基因及肌动蛋白(actin)α1受烯丙异噻唑诱导表达,部分克隆为低丰度转录本．     

Cloning and expression of a cDNA encoding ribosomal protein S4 from Rice (Oryza sativa)

A cDNA clone, pS4, has been isolated from a cDNA library prepared from rice anthers of about 1.0 mm in length. DNA sequence analysis and database search show that the cDNA encodes a protein which is highly homologous to eukaryotic 80s ribosomal protein subunit 4 (S4). Northern hybridization indicates that this gene expresses in all tissues analyzed although the expression level varies and it cannot be induced by mechanical wounding in leaves. Southern blot analysis demonstrates that this riceS4 gene is from a multigene family.     

Palynological records of environmental changes in the middle part of Inner Mongolia,China

Today, the central part of Inner Mongolia has a continental climate, with cold winters, hot summers, and approximately 70% of the precipitation falling in the summer months. Vegetation is dominated by steppe vegetation with Stipa bungeana and Stipa breviflora. Scattered individuals of Quercus liaotungensis, Populus davidiana, Betula platyplylla are distributed in the mountains, with some Pinus tabluaoformis, near Miaoyu, and with fragmentary Picea above 1700—1800 m a.s.l. The Huitenglian…     

An anther-specific chalcone synthase-like geneD5 related to rice pollen development

It was shown in a previous analysis that D5 gene from rice (Oryza sativa L.) was an anther-specific gene encoding a chalcone synthase-related protein. In this study, D5 gene was found specifically expressed in tapetum cells as well as in the peripheral cells of the vascular bundle of rice anthers by RNA in situ hybridization. In order to study its function, D5 was transformed into rice in both sense and antisense directions driven by a rice Actin 1 promoter. It has been observed that the pollen grains from the antisense D5 transgenic rice plants are abnormal, indicating that D5 plays a critical role in rice pollen development.     

光敏核不育水稻花粉败育中淀粉和蛋白质含量的研究(英文)

在可育花粉的发育中,功能叶同化产物运往花药合成淀粉和蛋白质,淀粉含量增加显著,颖壳和花药可溶性糖减少。长日下功能叶生理机能并未下降,糖也可运往颖花,但在颖壳中积累,不育花药中淀粉含量增加不明显,可溶性糖含量略有下降。单核晚期前可育花药蛋白质含量先是下降,但双核期后又迅速上升;不育花药中则基本上一直呈下降趋势。游离氨基酸总量与蛋白质含量变化相吻合,脯氨酸含量在可育和不育花粉之间差异不大。作者认为,淀粉和蛋白质的合成障碍是长日诱导光敏核不育水稻花粉败育的基本原因,这种障碍是由其底物供应不足引起的;游离氨基酸的变化与花粉败育仅有间接关系。     

Isolation of cDNA fragment of fertility-related gene in photoperiod-sensitive genic male sterile rice

The photoperiod_sensitive genic male sterile rice (PGMR) is particularly useful to take advantage of heterosis in rice. mRNA differential display was used to isolate the fertility_relative genes in rice. After establishing an optimized mRNA differential display system, one of the differential cDNA fragments that maybe related to the development and maturation of rice panicle was cloned from a PGMR Nongken 58S.     

Novel gene expressed during early embryogenesis of zebrafish identified by mRNA differential display

Following the revelation of the molecular mechanism of morphogenesis in fruitfly, research on the molecular mechanism of morphogenesis in vertebrate becomes the focus of developmental biology. The isolation of genes controlling the embryogenesis of zebrafish, a vertebrate model animal, is considered as an initial step toward investigating this issue. There are several approaches that can be used to isolate developmental genes, each of which is suited to a particular situation. In this note, mRNA differential display was utilized to demonstrate the mRNA differences among zebrafish embryos at 4, 5 and 6 h post fertilization (28.5℃, corresponding to oblong, dome and shield stages, respectively, called blastula, gastrula and neurula in this note). One cDNA tag that was specific to embryos at neurula stage was cloned and sequenced. After sequence comparison in Genbank, we found that this cDNA tag represents a novel gene. The expression of this gene in the developing zebrafish embryos was examined by whole mount in situ hybridization. The hybridization results confirmed that this gene was specifically expressed in zebrafish neurula embryos.     

Structural and expressional analysis of a cDNA that expresses predominantly in rice stamens

The development of stamen is an important stage during the sexual reproduction in higher plants. Studying the genes that predominantly or even specifically expressed in stamens will aid in undestanding the molecular mechanism of stamen development. The structure of a rice CHS-like gene (D5) which predominantly expressed in anthers has been analysised, showing high structural similarity with two other anther-specific genes (A1 and BA42) fromBrassica napus. Northern analysis also reveals that the three genes share a similar expression pattern and that the expression of D5 may relate to the early development of pollens. It is concluded that D5, A1 and BA42 represent a group of diverged CHS-like genes which are related to stamen development.     

水稻花药发育的细胞形态学精细分期

本研究通过观察籼型杂交水稻"川优9527"的花粉和药室壁的发育,对幼穗花药从雄蕊原基形成至花粉成熟的全过程进行了分期。选用不同发育时期的水稻幼穗(长度约1.0~10.0 mm),以石蜡切片技术分别制作其花药装片,置于光学显微镜(N ikon DS-R i1-U2)下观察并拍照。根据细胞及组织形态特征的显著变化,将花药的发育进程精细划分为:孢原细胞期、二层药室壁期、三层药室壁期、花粉母细胞形成期(以上4期对应小孢子母细胞形成期)、花粉母细胞减数分裂初期、花粉母细胞减数分裂中期、花粉母细胞二分体时期、花粉母细胞四分体时期(以上4期对应小孢子母细胞减数分裂期)、小孢子早期、小孢子中期、小孢子晚期、二胞花粉时期、成熟花粉时期等13个阶段。     

Cloning and sequencing of cDNA fragment of gene of wheat (Triticum aestivum. L) induced by dehydration

cDNA fragment of the gene (dehydration induced,di1) of wheat (Triticum aestivum. L) induced by 30% PEG-6000 (−1.13 MPa) treatment was isolated with mRNA differential display technique. Northern blot analysis showed that the expression ofdi1 gene improved at 10 h reached the highest at 48 h under 30% PEG-6000 treatment. cDNA fragment ofdi1 gene has been cloned and sequenced (211 bp). DNA sequence analysis shows that there is no homologue in GenBank todi1 cDNA.