摘要
建立抗感水稻品种受稻瘟菌侵染和未侵染蛋白质的双向凝胶电泳图谱,分析其差异表达的蛋白,寻找稻瘟病的抗性相关蛋白,以阐明稻瘟病的发病机制。采用TCA/丙酮沉淀法提取四种愈伤组织材料的总蛋白并采用固相pH梯度(immobilizedpHgradient,IPG)双向凝胶电泳(two-dimensionalgelelectrophshiya,oresis,2-DE)分离四种材料总蛋白质,凝胶经银染显色后,用PDQuest图像分析软件进行比较分析、识别差异表达的蛋白质。成功获得抗感水稻品种受稻瘟菌侵染和未侵染蛋白质的双向凝胶电泳图谱。获得未侵染内香优2号平均蛋白点数为447个,汕优63平均蛋白点数为440个;侵染后抗性品种内香优2号平均蛋白数为523个,感性品种汕优63平均蛋白质点数为326个。内香优2号未经侵染和侵染后图谱匹配率达89%和87%,汕优63未经侵染和侵染后图谱匹配率达86%和85%。内香优2号的差异表达蛋白点数为76个,汕优63的差异表达蛋白点数为114个。两者间存在一些差异表达的蛋白质,为阐明稻瘟病的发病机制打下了坚实的基础。
Pyricularia grisea is one of the most serious disease of rice in the world. To understand the mechanism of the resistance of rice against Magnaporthe grisea, differential proteomic comparison between resis tant and susceptible variety before and after Magnaporthe grisea infestation was conducted using 2 dimensional （2-D） gel electrophoresis to find the different resistance-related proteins of Magnaporthe grisea. The total proteins of four kinds of callus inductions were isolated by TCA/cold acetone precipitation. And these were prepared by means of immobilized pH gradient （IPG） based on two-dimensional gel electrophoresis. After silver staining, the differential expression proteins of four kinds were analyzed using image analysis software （PDQuest）. Using proper method stated above, satisfactory 2DGE maps of total proteins obtained and the preliminary analysis results were reported. The results showed that before and after Magnaporthe grisea infestation, average numbers of resistant variety protein spots were 447 and 523, and the corresponding average matching rate were 89% and 87%. The average numbers of susceptible variety protein spots were 440 and 326, and the corresponding average matching rate were 86% and 85%. The 76 differential protein spots were identified between the two types of resistant variety tissues, and the 114 differential protein spots were identified between those of susceptible variety tissues. The differentially expressed proteins can be observed in resistant and susceptible rice to Magnaporthe grisea, which can be used to elucidate the pathogenesis of Pyricularia grisea for further study. The research results suggested that the changes of protein after Magnaporthe grisea infestation might be the base of resistibility inside the rice.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2006年第12期92-98,共7页
China Biotechnology
基金
四川省青年科技基金项目(07JQ0034)
关键词
水稻
稻瘟菌
双向电泳
稻瘟病
愈伤组织
Rice Magnaporthe grisea Two-dimensional electrophoresis Rice blaste Allus induction
