栽培小麦Brock和京411感染白粉菌后蛋白质组的变化

doi:10.3724/SP.J.1006.2009.02064

作物学报 ›› 2009, Vol. 35 ›› Issue (11): 2064-2072.doi: 10.3724/SP.J.1006.2009.02064

栽培小麦Brock和京411感染白粉菌后蛋白质组的变化

于振¹,李倩¹,赵建叶¹,江帆¹,王振英^1,*,彭永康^1,*,解超杰²,刘志勇²,孙其信²,杨作民²

1天津师范大学化学与生命科学学院/细胞遗传与分子调控天津市重点实验室，天津300387；2中国农业大学农学与生物技术学院，北京100193

收稿日期:2009-02-12 修回日期:2009-05-08 出版日期:2009-11-12 网络出版日期:2009-09-10
通讯作者: 王振英, E-mail: wzycell@yahoo.com.cn; 彭永康, E-mail: pykcell@yahoo.com.cn
基金资助:
本研究由国家自然科学基金（30771346），天津市科委重点项目（08JCZDJC16500），天津市教委重点项目（2006ZD08）资助。

Proteome Changes in Wheat Varieties Brock and Jing 411 after Inoculating Blumenia graminis

YU Zhen¹,LI Qian¹,ZHAO Jian-Ye¹,JIANG Fan¹,WANG Zhen-Ying^1,*,PENG Yong-Kang^1*,XIE Chao-Jie²,LIU Zhi-Yong²,SUN Qi-Xin²,YANG Zuo-Min²

1College of Chemistry and Life Science/Tianjin Key Laboratory of Cyto-Genetical and Molecular Regulation,Tianjin Normal University,Tianjin 300387,China;2College of Agronomy and Biotechnology,China Agricultural University,Beijing 100193,China

Received:2009-02-12 Revised:2009-05-08 Published:2009-11-12 Published online:2009-09-10
Contact: WANG Zhen-Ying, E-mail: wzycell@yahoo.com.cn;PENG Yong-Kang, E-mail: pykcell@yahoo.com.cn

摘要/Abstract

摘要：

用华北地区流行的白粉菌15号生理小种，感染强抗白粉病的栽培小麦Brock和对白粉病敏感的小麦京411，通过蛋白质组技术分析其差异蛋白。结果表明，Brock经白粉菌感染12 h后，至少有6个蛋白质斑点(43 kD/pI 6.7、43 kD/pI 6.9、43 kD/pI 7.2、28 kD/pI 5.8、26 kD/pI 5.5和26 kD/pI 6.5)表达量明显增加；感染3 d后，有5个蛋白质斑点(48 kD/pI 5.6、43 kD/pI 6.9、43 kD/pI7.2、28 kD/pI5.8和26 kD/pI5.5)表达量增加；感染5 d后，有12个新的蛋白质斑点(16 kD/pI 7.6、42 kD/pI 6.5、40 kD/pI 4.8、40 kD/pI 4.6、31 kD/pI 5.7、16 kD/pI 4.6、20 kD/pI 8.3、50 kD/pI 6.7、48 kD/pI 6.6、28 kD/pI 5.7、23 kD/pI 4.8和25 kD/pI 4.7 )被诱导合成，2种蛋白质斑点(26 kD/pI 4.6和17 kD/pI 7.9)消失。京411经白粉菌感染12 h后，3个蛋白质斑点(21 kD/pI 6.4、18 kD/pI 5.4和14 kD/pI 7.0)表达量增加；感染3 d后，有2个蛋白质斑点(80 kD/pI 5.4和14 kD/pI 7.0)表达量增加，1个蛋白质斑点(16 kD/pI 5.4)表达量下降；感染5 d后，有3个蛋白质斑点(50 kD/pI 7.3、40 kD/pI 7.3和24 kD/pI 7.2)表达量增加，2个斑点(40 kD/pI 4.8和14 kD/pI 7.2)表达量下降，但没有发现新的蛋白质合成。对Brock中诱导产生的12个新蛋白质斑点，利用MALDI-TOF-MS方法，于NCBI进行数据查询，其中有6个分别属于F-box亮氨酸高度重复蛋白、重金属转运/解毒蛋白、β-1,3-葡聚糖酶(两个同工体)、β-1,3-葡聚糖酶前体、锌指蛋白。功能查询表明，上述6个蛋白参与细胞周期调控、发育、激素响应、基因转录和病害防御等。推测Brock和京411感染白粉菌后，出现的蛋白质组变化可能与各自的抗、感白粉病特性有关。

关键词: 白粉病, 小麦, 蛋白质组, ß-1,3-葡聚糖酶, MALDI-TOF-MS

Abstract:

The protein of resistant wheat (Triticum aestivum L.) variety Brock and susceptible variety Jing 411 was extracted from leaves and separated using two-dimensional polyacrylamid gel electrophoresis (2-DE) at 12 h, 3 d, and 5 d after inoculating prevalent race No. 15 of B. graminis f.sp. tritici. In Brock, compared with contral with no pathogen inoculation, at least six protein spots of 43 kD/pI 6.7, 43 kD/pI 6.9, 43 kD/pI 7.2, 28 kD/pI 5.8, 26 kD/pI 5.5 and 26 kD/pI 6.5 obviously increased in content at 12 h time point; and five protein spots of 48 kD/pI 5.6, 43 kD/pI 6.9, 43 kD/pI 7.2, 28 kD/pI 5.8 and 26 kD/pI 5.5 increased in content at 3 d time point. At 5d after inoculation, 12 novel proteins were induced, viz. 16 kD/pI 7.6, 42 kD/pI 6.5, 40 kD/pI 4.8, 40 kD/pI 4.6, 31 kD/pI 5.7, 16 kD/pI 4.6, 20 kD/pI 8.3, 50 kD/pI 6.7, 48 kD/pI 6.6, 28 kD/pI 5.7, 23 kD/pI 4.8 and 25 kD/pI 4.7; simultaneously, two protein spots that were observed earlier disappeared. In Jing 411, three protein spots of 21 kD/pI 6.4, 18 kD/pI 5.4, and 14 kD/pI 7.0 increased in content at 12 h after inoculation. At the 3 d time point, two protein spots of 80 kD/pI 5.4 and 14 kD/pI 7.0 increased in content, however, one protein spot (16 kD/pI 5.4) showed decrease in protein abundance. At the 5 d time point, three protein spots of 50 kD/pI 7.3, 40 kD/pI 7.3, and 24 kD/pI 7.2 and two protein spots of 40 kD/pI 4.8 and 14 kD/pI 7.2 showed increase and decrease in protein abundance, respectively, but there were no novel protein spots induced. Among the 12 novel protein spots induced in Brock, six spots were identified using MALDI-TOF-MS and NCBI database searching, which were F-box and leucine-rich repeat protein, heavy metal transport/detoxification protein, endo-beta-1,3-glucanase (two isozymes), beta-1,3-glucanase precursor and zinc finger protein. These proteins are involved in a wide range of physiological processes, such as cell cycle control, development, phytohormone response, and resistance to fungal disease. Thus, the proteome changes in Brock and Jing 411 leaves are probably associated with the resistance and susceptibility to powder mildew, respectively.

Key words: Powdery mildew, Wheat, Proteome, Beta-1,3-glucanase, MALDI-TOF-MS

于振,李倩,赵建叶,江帆,王振英,彭永康,解超杰,刘志勇,孙其信,杨作民. 栽培小麦Brock和京411感染白粉菌后蛋白质组的变化[J]. 作物学报, 2009, 35(11): 2064-2072.

YU Zhen,LI Qian,ZHAO Jian-Ye,JIANG Fan,WANG Zhen-Ying,PENG Yong-Kang,XIE Chao-Ji. Proteome Changes in Wheat Varieties Brock and Jing 411 after Inoculating Blumenia graminis[J]. Acta Agron Sin, 2009, 35(11): 2064-2072.

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栽培小麦Brock和京411感染白粉菌后蛋白质组的变化

Proteome Changes in Wheat Varieties Brock and Jing 411 after Inoculating Blumenia graminis

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