Understanding resistant germplasm-induced virulence variation through analysis of proteomics and suppression subtractive hybridization in a maize pathogen Curvularia lunata
Understanding resistant germplasm-induced virulence variation through analysis of proteomics and suppression subtractive hybridization in a maize pathogen Curvularia lunata
复制标题
通过玉米病原体新月弯孢菌的蛋白质组学分析和抑制消减杂交了解抗性种质诱导的毒力变异
DOI：
10.1002/pmic.201200105
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发表时间：
2012-12-01
期刊：
影响因子：
3.4
通讯作者：
Chen, Jie
中科院分区：
文献类型：
--
作者：
Gao, Shigang;Liu, Tong;Chen, Jie
玉米弯孢菌是引起玉米弯孢菌叶斑病的重要病原菌。在C. lunata。为了更好地理解这一现象的机制，我们连续将抗性玉米群体的选择压力施加到C。lunata菌株WS 18（低毒力）。结果，该菌株的毒力显著增强。使用2DE，12个上调和4个下调的蛋白质被确定在毒性增加的菌株相比，WS 18。我们的分析表明，黑色素合成相关蛋白（Brn 1，Brn 2和scytalone酶）和应激耐受相关蛋白（HSP 70）作为重要的标记或因子直接参与了C. lunata。为了验证2DE结果并在mRNA水平上筛选差异基因，我们构建了一个消减cDNA文库（测试者：毒力增加菌株;驱动者：WS 18）。共获得188个单基因，其中14个单基因是致病力进化的指示基因。Brn 1和热休克蛋白基因表现出相似的表达模式，相应的蛋白质检测2DE。总之，我们的研究结果表明，差异蛋白或基因，参与黑色素合成或胁迫耐受反应，可以被认为是毒力增加的标志。lunata。
Curvularia lunata is an important pathogen causing Curvularia leaf spot in maize. Significant pathogenic variation has been found in C. lunata. To better understand the mechanism of this phenomenon, we consecutively put the selective pressures of resistant maize population on C. lunata strain WS18 (low virulence) artificially. As a result, the virulence of this strain was significantly enhanced. Using 2DE, 12 up-regulated and four down-regulated proteins were identified in virulence-increased strain compared to WS18. Our analysis revealed that melanin synthesis-related proteins (Brn1, Brn2, and scytalone dehydratase) and stress tolerance-related proteins (HSP 70) directly involved in the potential virulence growth as crucial markers or factors in C. lunata. To validate 2DE results and screen differential genes at mRNA level, we constructed a subtracted cDNA library (tester: virulence-increased strain; driver: WS18). A total of 188 unigenes were obtained this way, of which 14 were indicators for the evolution of pathogen virulence. Brn1 and hsp genes exhibited similar expression patterns corresponding to proteins detected by 2DE. Overall, our results indicated that differential proteins or genes, being involved with melanin synthesis or tolerance response to stress, could be considered as hallmarks of virulence increase in C. lunata.