Role of the NBS-LRR Gene DEPG1, and the Two Receptor-Like Cytoplasmic Kinase Genes NRRB and XCRK in Rice - Bacterial Leaf Streak Pathogen Interactions

在世界一些地区尤其是亚洲热带和亚热带地区，细菌条斑病（BacterialLeafStreak，BLS）是水稻生产的一个重要限制因素。抗病育种是防治BLS的首选方法。用蛋白质组学的方法分析细菌条斑病菌（Xanthomonasoryzaepv.oryzicola，Xoc）侵染的水稻叶片蛋白组表达的差异，从中鉴定差异表达的蛋白基因，是一种研究水稻与细菌性条斑病菌Xoc互作分子机制的重要方法，有望从中鉴定出参与水稻和Xoc互作的重要蛋白，包括参与水稻防御反应的重要蛋白，进而获得相应编码基因。本论文是在获得上述差异表达蛋白基因的基础上，对其中1个NBS-LRR类基因DEPG1和2个类受体激酶基因NRRB...Rice bacterial leaf streak (BLS) caused by Xanthomonas oryzae pv. Oryzicola (Xoc) is a major biotic constraint to rice production in some areas of the world, especially the tropic and subtropical regions of Asia. Development of resistant cultivars is the preferred means of controlling this disease. Identification of differentially expressed protein genes from the rice proteomes under Xoc attack us...学位：理学博士院系专业：生命科学学院生物学系_生物化学与分子生物学学号：2162007015379

Preliminary Study on Regulation of XIOsPR10 Gene Expression in Rice

为探索水稻Pr10蛋白在水稻抗细菌性条斑病中的作用,构建了XIOSPr10基因的植物过量表达载体P1301-XIOSPr10及rnAI表达载体PdS1301-XIOSPr10,通过农杆菌介导分别转化水稻愈伤组织,获得了相应的再生植株。经guS检测和PCr分析,证实XIOSPr10基因以及rnAI片段分别整合到水稻再生植株基因组中;半定量rT-PCr分析显示,过量表达植株中XIOSPr10基因的表达量高于对照,而rnAI转基因植株中XIO-SPr10基因的表达被抑制。In order to study the function of rice XIOsPR10 gene which is related to the plant disease resistant reaction,we constructed plant over expression and RNAi expression vector and integrated into the genome of rice via Agrobacterium tumefaciens EHA105,respectively.The transgenic cultivars were identified by PCR and GUS gene expression detection.The analysis result showed that the transcription level of XIOsPR10 of over-expression transgenic rice was higher than that of control.In construct,XIOsPR10 gene expression was blocked in RNAi transgenic rice.科技部863专题(2007AA10Z132);国家重大科技专项(2008ZX08001-001;2009ZX08009-045B);教育部重点项目(01102

Studies on Rice Resistance-related Proteins in Response to Bacterial Leaf Streak,Xanthomonas Oryzae pv.Oryzicola

水稻细菌性条斑病由XAnTHOM OnAS OryzAEPV.OryzICOlA引起,是目前威胁亚洲稻区水稻生产的重要病害之一。以水稻品种9311为研究对象,通过差异蛋白质组学方法,研究了病菌侵染48 H后水稻叶片的差异表达蛋白质,通过分析比对、质谱分析以及数据库检索,从中选择了7个上调表达的鉴定蛋白,包括4个水稻lrk类基因,2个nbS-lrr类基因和1个Pr-10基因家族成员基因。并设计相应的PCr引物,从水稻CdnA中获得相关基因片段做探针进行nOrTHErn杂交分析,结果显示,上述基因在接种病原菌12 H或48 H后表达量增加,表明这些蛋白质参与了抗病反应。Rice bacterial leaf streak(BLS) caused by the pathogen Xanthomonas oryzae pv.oryzicola(Xooc) is one of the major diseases in Asia.In this paper,the protein differently expressed after infected for 48 h by BLS was studied with rice 9311 as material through proteomic approach.Based on the result of protein blast,fingerprints and data search analysis,seven identified proteins up-regulated were selected including four LRK like genes,two NBS-LRR type genes and one PR-10 gene.Relevant primers were designed to amplify the genes from rice cDNA as probe for Northern blotting.The results showed that the mRNA levels of these genes increased significantly after infected by BLS for 12 h and 48 h,indicating these proteins have participated in disease resistance.国家863计划项目(2007AA10Z132);国家重大科技专项(2008ZX08001-001;2009ZX08009-045B);教育部重点项目(重点01102)资

Transformation of a NBS-LRR Protein Gene and Its Overexpression in Rice

水稻OSXOC1334是一个编码nbS-lrr类蛋白质的基因,通过构建该基因的超量表达载体,并在农杆菌EHA105介导下转化中花11号水稻,最终获得6株再生水稻植株.对抗性愈伤组织和再生植株用guS组织化学染色检测,结果抗性愈伤组织和6株再生水稻植株均可染上蓝色,而野生型植株不变色,用潮霉素磷酸转移酶基因的特异引物进行PCr鉴定,再生植株均可扩增到目标条带而野生型植株则不能,表明OSXOC1334基因已随T-dnA整合到再生水稻植株基因组中.采用半定量和荧光定量PCr分析转基因水稻的OSXOC1334基因表达,结果其中3株转基因植株的OSXOC1334基因平均相对表达量明显增强,其中1株为野生型植株的23.5倍.这为进一步鉴定该基因功能奠定了基础.Osxoc1334 was a NBS-LRR protein gene.Its overexpression vector was constructed and transformed rice variety Zhonghua11(Oryza sativa ssp.japonica) through Agrobacterium tumefaciens EHA105 mediated system,and thus six regenerated plantlets were obtained.The regenerated resistant embryogenic calli and plants were tested by GUS histochemical staining.As a result,all the regenerated calli and plants turned into blue but non-regenerated calli and wild type plants.These regenerated plantlets were also identified by PCR using hygromycin phosphotransferase gene specific oligonucleotide primers.The anticipated DNA fragments were amplified from all regenerated plants but wild type plant in the end.These data suggested that Osxoc1334 gene was introduced into the genome of the regenerated plants with T-DNA.The expressional levels of Osxoc1334 gene in these T0 plants were analyzed by semiquantitative RT-PCR and real time PCR.The expressional levels of Osxoc1334 gene in three T0 plants were enhanced intensively.The average relative expressional level of Osxoc1334 gene in one T0 plant reached about 23.5 times as compared to wild type plants.科技部863专题(2007AA10Z132;2008ZX08001-001;2009ZX08009-045B);教育部科学技术研究重点项目(01102

Prokaryotic Expression and RNase Activity Analysis of Rice XIOsPR10

Pr10(病程相关蛋白10)类蛋白与植物的抵御外来病害及系统获得性抗性(SAr)有着紧密联系,而且许多Pr10类蛋白都具有rnASE活性,并通过这种活性抵抗外源病害。根据获得的XIOSPr10基因,将其构建到PET28A中,通过原核表达及磁珠纯化获得目的蛋白,通过rnA消化实验证实XIOSPr10重组蛋白具有rnASE活性,进一步揭示了XIOSPr10蛋白的功能。It has been regarded that PR10s played an important role in systemic acquired resistance(SAR).Furthermore,many PR10 proteins have been reported to exhibit RNase activity and predicted to be responsible for its antibiotic activity.So the Prokaryotic Expression Vector pET28a-XIOsPR10 was constructed and transferred to BL21(DE3).The recombinant protein XIOsPR10 was obtained by Prokaryotic expression and purification of magnetic microbeads,and showed the RNase activity.国家“863”计划(2007AA10Z132);国家科技重大专项(2008ZX08001-001、2009ZX08009-045B);教育部科技研究重点项目(01102)资助项

遥感技术在塔里木河流域自然资源调查中的应用

该项课题应用航空遥感为主，结合多种信息资料，首次对我国最长的内陆河——塔里木河两岸进行资源与环境的综合性调查，完成了地貌、水系、土地类型、土地资源、土地利用、沙漠化、植被、胡杨林、草地等９种专题系列图件，共编制１∶１０万标准图幅专题图３１４幅，并编写了说明书，量算了资源、环境类型面积；获取了一套较新和系统完整的资源、环境数据；探讨了遥感应用的新方法，为合理制订流域规划和开发利用塔里木河的水资源等提供了依据