立枯丝核菌RsCAT基因的克隆及在菌核形成中的表达分析
为了明确过氧化氢酶(catalase, CAT)在立枯丝核菌菌核形成中的作用,在玉米纹枯病菌Rhizoctonia solani AG-1-IA中克隆了CAT 基因,利用生物信息学软件推测其理化性质,结构域,二级结构并构建该蛋白的三维模型,系统进化树分析表明该蛋白与Laccaria bicolor同源性最高。利用real-time qPCR 分析RsCAT在正常情况下和H2O2 胁迫条件下的表达差异。结果表明,RsCAT在菌核形成过程中表达量持续升高,在第6天时表达量最高然后开始下降,在H2O2 胁迫条件下,处理组CAT 表达量随着时间的延长逐渐提高,其基因表达量,菌丝生长速率和菌核的生物量均高于对照组。综上所述,初步判断RsCAT在玉米纹枯病菌菌核的形成中起到清除H2O2等自由基的作用。
英文摘要:
In order to understand the role of catalase in the process of sclerotium formation of Rhizoctonia solani, the CAT gene was cloned in Rhizoctonia solani AG-1-IA. The bioinformatics tools was used to predict its physical and chemical properties, domain, secondary structure and construct the three-dimensional model of the protein. Phylogenetic tree analysis showed that the protein had the highest homology with Laccaria bicolor. Real-time qPCR was used to analyze the expression of CAT in the process of sclerotium formation and under H2O2 stress. The results showed that the expression of RsCAT increased during the process of sclerotium formation, and the expression reached the highest level on the 6th day and then began to decline., The expression of CAT in the treatment group increased gradually with the prolongation of time under the H2O2 stress , the gene expression, mycelial growth rate and sclerotium biomass were higher than those in the control group. In summary, the preliminary judgment of RsCAT in the formation of scavenging H2O2 and other free radicals.
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