【作者】 黄青云；

【导师】 陈亮；

【作者基本信息】 厦门大学 ， 细胞生物学， 2006， 硕士

【摘要】 水稻细菌性条斑病(简称细条病),广泛分布于华南、中南稻区。该病是细条病菌Xanthomonas oryzae pv. oryzicola (Xooc)引起的一种细菌性病害,于20世纪50年代在我国广东省首先发现,后因感病的杂交水稻大面积推广,致使此病迅速蔓延,病区不断扩大,已成为我国水稻主要病害和重要的检疫对象。目前国内外对于水稻细条病的研究仅仅限于基因组学的研究以及抗性基因的转化,而对该病菌侵染水稻后诱导机体蛋白应答的系统性研究还未见报导。因此,运用差异蛋白质组学手段开展水稻对细条病菌侵染的应答相关蛋白研究,无论是在阐明水稻的抗性分子机理和基因表达调控方面,还是在高产、优质、高抗的水稻品种的分子育种方面具有重要意义。本试验以水稻品种“明恢63”和强毒力细条病菌株“89773-1-1”为供试材料,在水稻主茎六叶一心期时,提取叶片全蛋白运用双向凝胶电泳分离,研究了叶片在接菌处理和不接菌的对照条件下不同时期蛋白质组差异表达谱,质谱分析鉴定差异蛋白质,通过对差异表达蛋白质的分析,探讨水稻对细条病菌侵染的防卫应答系统。在细菌侵染后2 h、5 h、12 h、48 h 4个处理时间梯度进行取样,提取叶片全蛋白样品,通过双向电泳-质谱技术,比较其蛋白质表达情况。结果发现,共有33个蛋白质点在不同侵染时期发生了差异表达,其中表达量上升的有21个;表达量下降的有12个。经MALDI-TOF/MS检测及肽指纹图谱(PMF)分析,其中27个蛋白点在有关数据库中得到了归属鉴定。进一步对这27个蛋白及其特性进行综合分析,认为这些蛋白参与了水稻对细条病侵染的信号识别及防卫应答,其中包括信号转导类蛋白、防卫相关蛋白、代谢相关蛋白和参与蛋白质合成的蛋白等。结果表明,中感水稻品种“明恢63”对细条病病菌侵染后的生物应答受复杂的网络调控机制控制。这些功能蛋白的鉴定为探明水稻对细条病的抗性机理,明确细条病菌与寄主植物水稻相互作用的分子基础及其抗病信号转导途径及寻找与抗细条病菌密切相关的差异表达蛋白质和基因奠定了基础。更多还原

【Abstract】 Rice bacterial leaf streak (BLS) distributes widely in South China. BLS which was caused by Xanthomonas oryzae pv. oryzicola (Xooc) firstly discovered in the 1950’s in Guangdong, and then spread quickly because the infective hybridized rice was popularized. So BLS is one of the major rice diseases in South China and becomes an important subject of quarantine in our country. Presently, the research of rice BLS is only limited in the research of genomics and the transformation of resistant gene, while very few systemic research on the response to BLS at the level of proteins have been reported.So it is greatly significant to investigate the responses proteins to BLS by means of differential proteomics, either in illuminating the molecular mechanism of rice resistance and the control to the express of gene, or the molecular breeding of the rice with high yield, high quality, and high resistance.The materials in this experiment are rice cultivar minghui63 (Oryzae sativa L.) and“89773-1-1”, the pathogen Xooc strong pathogenicity in South China. The rice bacterial leaf streak was inoculated to the leaves in six-leaf stage and the leaf proteins were extracted. The defense-response system of rice against Bacterial Leaf Streak was probed into by the analysis of the differentially expressed proteins of Minghui 63 leaf after BLS infection in different inoculation time with the methods of two-dimensional electrophoresis and mass spectrometry.Proteins were extracted from leaves 2, 5, 12, and 48 hours after rice Bacterial Leaf Streak inoculation and separated by two-dimensional (2-D) polyacrylamide gel electrophoresis. 33 proteins resolved on the 2-DE gels were induced or increased in the inoculated leaf. 21 proteins were up-regulated, while 12 proteins were down- regulated. 27 proteins were determined by matrix - assisted laser desorption/ionization-time of flight mass spectrometry, and identified protein database matching. Further analysis of these 27 proteins showed them involved in signal recognition and defense response, including signal transduction proteins, defense associated proteins, metabolism associated proteins and protein synthesis更多还原