氮饥饿环境中稻瘟病不同致病力菌株分泌蛋白的双向电泳图谱分析

Comparison analysis of Secretary Proteins Induced by Nitrogen Starvation of Different Pathogenicity Magnaporthe grisea Isolates by Two-Dimensional Gel Electrophoresis

采用双向电泳技术研究了稻瘟病菌株Y99-63和Y98-16在氮饥饿条件下分泌蛋白群的组成及差异。在稻瘟病菌株Y99-63和菌株Y98-16分泌蛋白的2-DE图谱中分别检测出262±10和253±10个蛋白质点。其中有130个蛋白质点为菌株Y99-63特异表达,121个蛋白质点为Y98-16特异表达,132个蛋白质点为两菌株共同表达。在两菌株共同表达的表达量相差达5倍以上的有38个蛋白质点,其中有25个蛋白质点在菌株Y99-63表达量较大,有13个蛋白质点在菌株Y98-16表达量较大。生物信息学预测稻瘟病菌分泌蛋白的分子量在10～20 kDa以及等电点在5.0～5.5范围内蛋白质点数最多。菌株Y99-63特异表达分泌蛋白的分子量10～20 kDa范围内蛋白质点最多,菌株Y98-16特异表达分泌蛋白的分子量在20～30 kDa范围内蛋白质点数最多,两菌株共同表达的分泌蛋白分子量在10～20 kDa范围内蛋白质点数最多。菌株Y99-63特异表达分泌蛋白的等电点在5.5～6.0范围内蛋白质点数最多,菌株Y98-16特异表达分泌蛋白的等电点在4.5～5.0范围内蛋白质点数最多,两菌株共同表达分泌蛋白等电点在5.0～5.5范围内蛋白质点数最多。稻瘟病菌不同菌株在氮饥饿条件下产生的分泌蛋白有较大差异。

We analyzed the secretary proteins induced by nitrogen starvation of Magnaporthe grisea from two different pathogenicity strains, i. e. Y99-63 and Y98-16 by two-dimensional electrophoresis （2-DE） to study their difference. Results showed that 262 ± 10 and 253± 10 protein spots on 2-DE maps were recordable based on default critics of PDQuest 7. 40, respectively. 130 spots in Y99-63 and 121 spots in Y98- 16 specifically presented on gels, and 132 spots were both detected in two strains. Among 132 spots, the expression level of 38 protein spots （25 protein spots in Y99-63 and 13 protein spots in Y98-16） were more than 5 times difference between two strains. The secretary proteins of the M. grisea was predicted using bioinformatics softwares as well, the molecular weight （MW） range of the largest number of protein spots that predicted by Bioinformatics softwares were distributed in 10 -20 kDa while the isoelectrie point （pI） range of these protein spots were mainly distributed in 5. 0 to 5. 5. The largest number of protein spots spe- cific expressed was located within 10 to 20 kDa range and 20 to 30 kDa range of MW for Y99-63 and Y98- 16, respectively, the largest number of protein spots co-expressed were located within 10 to 20 kDa range of MW. The largest number of protein spots specific expressed was located within 5. 5 to 6. 0 range and 4. 5 to 5. 0 range of pI for Y99-63 and Y98-16, respectively, the largest number of protein spots co-expressed were located within 5.0 to 5. 5 range of pl. The secretary proteins induced by nitrogen starvation of M. grisea in two strains were quite different on 2 DE-gel.