【作者】 林涛；

【导师】 陈亮；

【作者基本信息】 厦门大学 ， 发育生物学， 2005， 硕士

【摘要】 水稻细菌性条斑病是由Xanthomonas oryzae pv. oryzicola引起的,在热带、亚热带稻区广泛发生的细菌病害,也是目前威胁我国南方稻区水稻生产的重要病害之一,可以引起5%-40%的产量损失。本研究以对水稻细菌性条斑病极感病的水稻品种-9311为研究对象。通过双向电泳联用质谱技术,研究了9311在水稻细菌性条斑病病菌侵染后4个不同时间段(12h、48h、5d、10d)的叶片全蛋白2-DE图谱的变化,并对差异表达的蛋白质点进行MALDI-TOF/MS分析。2-DE图谱分析结果表明:在病原菌侵染9311后不同时期,共检测到164个差异蛋白出现,其中表达量上升的23个;表达量下降的16个;诱导出现的蛋白71个;抑制表达的蛋白54个。将164个差异蛋白点经MALDI-TOF/MS分析,得到其中106个蛋白的肽质量指纹图谱。通过MASCOT数据库比对,鉴定出水稻细菌性条斑病病菌侵染诱导差异表达蛋白86个。对这86个差异表达蛋白的同源性序列比对以及功能分析,并着重研究了其中的R和PR类蛋白,得到了其中NBS-LRR类蛋白质的同源进化树,以及NBS-LRR类蛋白和RLK/激酶类蛋白的保守区/功能区的分析结果。并根据蛋白序列分析结果,设计PCR引物,经RT-PCR,从水稻cDNA中,获得水稻RLK/激酶类基因片断5个,NBS-LRR类基因片断2个,PR-10基因家族成员基因片断1个。其中一个编号13-3-4的NBS-LRR类基因,在基因库中,与现有的同类基因最高的同源性为91%。经Northern验证,4-3-2基因在接种后12h,48h后表达量逐渐增大。本研究为水稻细菌性条斑病病原菌与寄主植物-水稻之间的侵染相互作用,以及寄主植物的防御应答反应网络研究提供了研究方更多还原

【Abstract】 Bacterial leaf streak (BLS), caused by Xanthomonas oryzae pv. Oryzicola, is widely distributed in tropical and subtropical area, and now emerging as an important pathogen of rice in southern China. Typically, BLS can cause grain weight losses range from 5-40%.The highly susceptible hybrid rice variety-9311 was used in all our experiments. Total proteins are extracted from leaves 12h, 48h, 5d, and 10d after rice BLS bacterium inoculation. We have used two dimensional electrophoresis (2-DE) and Mass spectrometry (MALDI-TOF/MS) to identify proteins that are differentially expressed in response of BLS infection. From 2-DE data, 164 resolved differentially expressed proteins were detected, 94 of those were induced, and 70 of those were repressed. Selected spots were subjected to tryptic digestion followed by identification using matrix-assisted laser desorption/ionization-time of flight-mass spectrometry. Total 86 proteins were identified. We got the polygenetic tree of NBS-LRR type proteins and the conserved domains of NBS-LRR-type and RLK/Kinase-like proteins. Basing on the result of protein sequences analysis, we designed the PCR probes and got 8 genes from cDNA of rice, including 5 RLK/Kinase-like genes, 2 NBS-LRR type genes and 1 PR-10 gene. One of the NBS-LRR type genes, protein No. 13-3-4, showed highest homology to the other same type genes was 91%.The study can direct the further research about pathogen-plant interaction and defense responses in rice. At the meantime, we analyzed 2 important kinds of response proteins, got 8 sequence fragments of更多还原