水稻稻瘟菌抗性相关蛋白的双向电泳分析

中国生物工程杂志

技术与方法

水稻稻瘟菌抗性相关蛋白的双向电泳分析

谢娟黄明星卢代华王胜华陈放

四川大学四川大学四川省农科院植保所四川大学四川大学

Analysis of Resistance-related Proteins in Rice Against Magnaporthe grisea by Two-dimensional Gel Electrophoresis

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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( immobilized pH gradient, IPG)双向凝胶电泳( two-dimensional gel electrophshiya, 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。

Abstract: To find the different resistance-related proteins of magnaporthe grisea by using two-dimensional gel electrophoresis for understanding the mechanism of the resistance of rice against magnaporthe grisea. The total proteins of four kinds of callus inductions were isolated by TCA/cold acetone precipitation. The four kinds of total proteins were prepared by means of immobilized pH gradient based on two-dimensional gel electrophoresis. After silver staining, gel-image analysis was performed by using PDQuest. Using proper method stated above, satisfactory 2DGE maps of total proteins obtained and the preliminary analysis results were reported. The results showed that average numbers of resistant variety protein spots were 447 and 523 before and after Magnaporthe grisea infestation, 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 the two types 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.

收稿日期: 2006-09-26 出版日期: 2006-12-25

通讯作者: 王胜华

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	黄明星
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引用本文:

谢娟,黄明星,卢代华,王胜华,陈放. 水稻稻瘟菌抗性相关蛋白的双向电泳分析[J]. 中国生物工程杂志, .

. Analysis of Resistance-related Proteins in Rice Against Magnaporthe grisea by Two-dimensional Gel Electrophoresis. China Biotechnology, .

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https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2006/V26/I12/92

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