水稻Xa7基因抗白叶枯病应答中的活性氧代谢分析

白叶枯病是由革兰氏阴性黄单孢菌水稻变种（Xanthomonas oryzae pv．Oryzae，Xoo）所引起的一种世界性水稻细菌病害．水稻Xa7基因是一个具有广谱抗性的显性抗白叶枯病基因．通过对水稻抗病品种IRBB7（含Xa7）和感病对照IR24接种白叶枯菌PX086，发现：在叶片的病原菌侵染部位，IRBB7比IR24的活性氧（H2O2和O2-）积累更快且含量更高；与活性氧代谢相关的酶，如超氧化物歧化酶、过氧化氢酶、抗坏血酸过氧化物酶和过氧化物酶的活性也更高．推测活性氧的代谢调节可能在Xa7基因介导的抗病反应中起作用．     

Isolation ofosRACD gene encoding a small GTP-binding protein from rice

Using an improved version of mRNA differential display technology, we have obtained a differentially displayed fragment RDP-8. Homologous comparison indicated that the fragment RDP-8 has high homology with the gene encoding maize small GTP-binding protein. By screening cDNA library of the rice Nongken 58N pan icle using the newly obtained fragment RDP-8 as probe, we further found the full-length cDNA of osRACD gene that encodes a rice small GTP-binding protein. Asco mpared with maize RACD gene, the osRACD of rice shows remarkable homology in both nucleotide sequence and amino acid sequence, 88% and 97% respectively. Evidence from RT-PCR study indicates that osRACD gene is related to photoperiod fertility conversion of photoperiod sensitive genic male sterility (PSGMS) rice.     

Cloning and characterization of rice RH3 gene induced by brown planthopper

Histones are basic low molecular weight proteins found in all eukaryotic genomes. The histones include five classes of basic proteins (H1, H2A, H2B, H3 and H4) that interact with each other and nuclear DNA to form the nucleosome. The H3 and H4 histone proteins are highly conserved and form the central tetrameric block of the core-nucleosome. Histone H3 has several post-transcrip- tional modifications such as methylation, acetylation, phosphonation, and ADP-ribosylation and it plays impor…     

Construction and characterization of a bacterial artificial chromosome library of rice 5460F

Thermo-sensitive genie male sterile (TGMS) rice has a number of desirable characteristics for hybrid rice production. Many studies have demonstrated that the sterility of TGMS rice is controlled by a single recessive gene. It has been mapped for the first time on chromosome 8 and namedtms 1. Several AFLP markers which tightly linked to thetms 1 gene have been identified recently. In order to develop a detailed physical map of thetms1 gene-encompassing region and finally clone thetms1 gene, a bacterial artificial chromosome (BAC) library of rice 5460F (the fertile mutant line of TGMS rice 5460S) using a modified vector pECBAC1 has been constructed. The constructed 5460F BAC library consists of 16 896 clones with an average insert size of 119 kb, which represents about 4.7 times rice haploid genome equivalents. Neither chloroplast nor mitochondrial DNA was detected from the library. The library was screened with three single copy sequence amplified fragment length polymorphism (AFLP) markers which tightly linked totms1 gene as probes and eight positive clones were identified.     

水稻紫色柱头突变体ps-5的遗传分析和分子定位

在水稻品种Nipponbare中发现一个紫色柱头突变体ps-5,整个生育期最显著的特点就是柱头紫色且子房较大.遗传分析表明该基因受1对隐性核基因控制,暂命名为ps.利用极端个体定位法把ps-5精细定位于第8染色体SSR标记LR41和LR43之间,ps-5基因距它们的遗传距离均为0.12cM,并与LR413共分离,LR41和LR43两标记间的物理距离约为125kb,为该基因的分子标记辅助选择和图位克隆奠定了基础.     

Production of bacterial blight resistant lines from somatic hybridization between Oryza sativa L. and Oryza meyeriana L

Novel bacterial blight (BB) resistance gene(s) for rice was (were) introduced into a cultivated japonica rice variety Oryza sativa (cv. 8411), via somatic hybridization using the wild rice Oryza meyeriana as the donor of the resistance gene(s). Twenty-nine progenies of somatically hybridized plants were obtained. Seven somatically hybridized plants and their parents were used for AFLP (amplified fragment length polymorphism) analysis using 8 primer pairs. Results confirmed that these plants were somatic hybrids containing the characteristic bands of both parents. The morphology of the regenerated rice showed characters of both O.sativa and O.meyeriana. Two somatic hybrids showed highest BB resistance and the other 8 plants showed moderate resistance. The new germplasms with highest resistance have been used in the rice breeding program for the improvement of bacterial blight resistance.     

Cloning and characterization of a glucose 6-phosphate／phosphate translocator from Oryza sativa

Plastids of nongreen tissues import carbon as a source of biosynthetic pathways and energy, and glucose 6-phosphate is the preferred hexose phosphate taken up by nongreen plastids. A cDNA clone encoding glucose 6-phosphate/phosphate translocator (GPT) was isolated from a cDNA library of immature seeds of rice and named as OsGPT. The cDNA has one uninterrupted open reading frame encoding a 42 kDa polypeptide possessing transit poptide consisting of 70 amino acid residues. The OsGPT gene maps on chromosome 8 of rice and is linked to the quantitative trait locus for 1000-grain weight. The expression of OsGPT is mainly re-stricted to heteretrephic tissues. These results suggest that glucose 6-phosphate imported via GPT can be used for starch biosynthesis in rice nongreen plastids     

Construction of the primary physical map of rice chromosome 12

A primary physical map of rice chromosome 12 was constructed using marker-based chromosome landing and chromosome walking. A BAC library from IR64 was screened using 84 RFLP markers, 4 STS markers and 6 microsatellite markers on chromosome 12 by colony hybridization and polymerase chain reaction (PCR) amplification. A total of 59 contigs consisting of 419 BAC clones including 5 single-clones were physically aligned on rice chromosome 12 with the largest BAC contig covering 855 kb. The whole physical map had a size of ∼16 Mb and covered about 52% of rice chromosome 12. This physical map will be certainly helpful for map-based gene cloning of agronomically and biological important genes and understanding the genome structure of the chromosome. Foundation item: Supported by Rockefeller Foundation Biography: FU Bin-Ying (1965-), male, Ph. D. candidate, Reseach direction: plant molecular genetics.     

拟南芥NCED3基因在水稻中的过量表达可以提高水稻的干旱胁迫耐受性

9-顺-环氧类胡萝卜素双加氧酶(NCED)是高等植物脱落酸(ABA)生物合成途径中的关键酶,其催化的裂解反应直接生成ABA的前体物质.本文应用拟南芥rd29A诱导型启动子和35S组成型启动子成功地将AtNCED3基因在水稻中过量表达,通过耐旱性筛选实验证明转基因水稻对干旱胁迫的耐受性有了显著提高,并且该优良性状在T2代中得到稳定遗传.进一步的分析表明,过量表达AtNCED3可以促进转基因水稻种子的休眠;在含rd29A诱导型启动子的转基因水稻中检测到ABA下游基因OsBZ8的表达;推测 AtNCED3在水稻中的过量表达可能会提高水稻中内源 ABA的水平,从而提高植株的耐旱性.     

Promoter of soybean early nodulin gene<Emphasis Type="Italic">enod2B</Emphasis> is induced by rhizobial Nod factors in transgenic rice

Nod factors, which are signaling molecules produced by Rhizobia, are the principal determinants of host specificity in Rhizobium-legume symbiosis. Nod factors can elicit a number of characteristic developmental responses in the roots of legumes, such as depolarization of the membrane potential in epidermal cells, specific expression of early nodulin genes and changes in the flux of calcium in root hairs, deformation of root hairs, cell division in the root cortex and formation of the nodule primordinm. Whether the rice plant can respond to signaling molecules (i.e. Nod factors) is an important question, as it could establish the potential for symbiotic nitrogen fixation in rice. The promoter of the soybean (Glycine max) early nodulin gene Gmenod2B fused to the β-glucuronidase (GUS) reporter gene was used as a molecular marker to explore whether Nod factors can be recognized by rice cells as signaling molecules. Transgenic rice plants harboring the chimeric gene Gmenod2BP-GUS were obtained via an Agrobacterium tumefaciens-mediated system. NodNGR factors produced by a broad-host-range Rhizobium strain NGR234(pA28) were used as probes to investigate the activity of the Gmenod2B promoter in rice. Our results showed that the early nodulin gene Gmenod2B promoter was induced by NodNGR factors in transgenic rice, and that it was specifically expressed in rice plant roots. Moreover, GUS gene expression driven by the Gmenod2B promoter in transgenic rice was regulated by nitrogen status. These findings indicated that rice possessed the ability to respond to Nod factor signals, and that this signal transduction system resulted in activation of the Gmenod2B promoter. Thus, we predict that the Nod-factor inducible nodulin expression system, which is similar to Rhizobium-legume symbiosis, may also exist in rice.     

分子标记辅助选育香型巨胚水稻

香型稻米因其在蒸煮和食用时都会有特殊的香味受到人们的喜爱．研究中,分别以实验室培育的香型正常胚水稻和非香巨胚水稻“上师大5号”作为亲本进行杂交,结合水稻香味基因分子标记辅助常规育种成功选育出香型巨胚水稻“上师大8号”．比较分析“上师大5号”和“上师大8号”两种巨胚水稻主要农艺性状和产量性状．结果显示：虽然“上师大5号”水稻平均每穗实粒数极显著地超过“上师大8号”水稻,但是“上师大8号”有效穗数略多于“上师大5号”水稻,而且千粒重也略高于“上师大5号”水稻,由此两者平均单株重接近,分别为29．10g和28．92g,统计分析差异不显著．两种巨胚水稻胚性状分析显示：“上师大8号”巨胚水稻的胚重量以及胚体积都与“上师大5号”巨胚水稻统计分析无显著差异,而且两种巨胚水稻的胚与糙米重量比以及胚与糙米体积比,统计分析都没有显著差异．开展此研究,为今后香型巨胚水稻的市场开发应用奠定了重要基础．     

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.     

Comparative mapping of QTLs for Al tolerance in rice and identification of positional Al-induced genes

Aluminum (A1) toxicity is the major factor limiting crop productivity in acid soils. In this study, a recombinant inbreed line (RIL) population derived from a cross between an A1 sensitive lowland indica rice variety IR1552 and an A1 tolerant upland japonica rice variety Azucena, was used for mapping quantitative trait loci (QTLs) for A1 tolerance. Three QTLs for relative root length (RRL) were detected on chromosome 1,9, 12, respectively, and 1 QTL for root length under A1 stress is identical on chromosome 1 after one week and two weeks stress. Comparison of QTLs on chromosome 1 from different studies indicated an identical interval between C86 and RZ801 with gene(s) for A1 tolerance. This interval provides an important start point for isolating genes responsible for A1 tolerance and understanding the genetic nature of Al tolerance in rice. Four A1 induced ESTs located in this interval were screened by reverse Northern analysis and confirmed by Northern analysis. They would be candidate genes for the QTL.     

Genetic analysis and fine mapping of an enclosed panicle mutant esp2 in rice （Oryza sativa L.）

The phenomenon of panicle enclosure in rice is mainly caused by the shortening of uppermost internode.Elucidating the molecular mechanism of panicle enclosure will be helpful for solving the problem of panicle enclosure in male sterile lines and creating new germplasms in rice.We acquired a monogenic recessive enclosed panicle mutant,named as esp2 (enclosed shorter panicle 2),from the tissue culture progeny of indica rice cultivar Minghui-86.In the mutant,panicles were entirely enclosed by flag leaf sheaths and the uppermost internode was almost completely degenerated,but the other internodes did not have obvious changes in length.Genetic analysis indicated that the mutant phenotype was controlled by a recessive gene,which could be steadily inherited and was not affected by genetic background.Apparently,ESP2 is a key gene for the development of uppermost internode in rice.Using an F 2 population of a cross between esp2 and a japonica rice cultivar Xiushui-13 as well as SSR and InDel markers,we fine mapped ESP2 to a 14-kb region on the end of the short arm of chromosome 1.According to the rice genome sequence annotation,only one intact gene exists in this region,namely,a putative phosphatidylserine synthase gene.Sequencing analysis on the mutant and the wild type indicated that this gene was inserted by a 5287-bp retrotransposon sequence.Hence,we took this gene as a candidate of ESP2.The results of this study will facilitate the cloning and functional analysis of ESP2 gene.     

New technology to examine and improve the purity of hybrid rice with herbicide resistant gene

The herbicide resistant bar gene has been widely used as selectable marker genes in the study on plant genetic transformation. Owing to the integration of the gene into rice genome, transgenic rice was resistant to the herbicide Basta. Therefore, selection of transgenic plant md genetic analysis became easier. In the studies, bar gene was introduced as a genetic marker gene into ria restore lie variety of two-line or three-line. Combined with conventional breeding method, the good herbicide resistant individual plant was derived as the new restorc line for hybrid combination. After sprayed with Basta at seeding phasc. the real hybrid rice that hnd herbicide resistance could grow normally, while the fnke hybrid rice and weeds, having no bar gene, were killed by herbicide. The method described above helped rapidly to examine and improve the purity of hybrid rice.     

水稻RSSG8基因启动子与GUS融合基因的构建及在烟草中的瞬间表达

使用染色体步移(Genome walking)法，从籼稻(Oryza sativa subsp．indica)桂朝2号基因组中克隆到长度为471bp的水稻精细胞优势表达基因RSSG8的启动子片段R8PN，并进行了全序列测定和分析．该段序列中含有3个CAAT-box，6个Gobox和多种植物顺式作用元件，但没有发现典型的TATA-box，推测为一种特殊的启动子结构，为了鉴定RSSG8基因的基本启动子元件，将二条长度不同的5’端侧翼区缺失体(分别长471bp，260bp)定向插入载体pBI121中，取代原有的CaMV 35S启动子，构了驱动报告基因GUS的植物表达载体pRGUS1，pRGUS2，通过农杆菌介导的瞬时表达法转化烟草叶片和花粉，快速鉴定启动子片段中起关键作用的区域，结果显示两个缺失片段都能启动GUS的表达，可以初步判定这两个片段具有启动子功能。     

水稻花期干旱胁迫应答基因Os07g0422100启动子的克隆及鉴定

水稻作为需水量最大的作物之一,对于水分的胁迫异常敏感.对处于花期的水稻进行干旱胁迫处理,利用基因芯片技术筛选出在水稻花穗中特异性表达的应答干旱胁迫的基因Os07g0422100.在对该基因的研究中发现,该基因的表达图谱有较高的专一性,且该基因的启动子属于诱导型启动子,仅在受到干旱胁迫的水稻花穗中表达.利用Os07g04...