中国农业科学Issue(15)：2971-2979,9.DOI:10.3864/j.issn.0578-1752.2014.15.007
灰葡萄孢BcKMO在病菌生长、发育和致病过程中的功能
Function Analysis ofBcKMOGene in Growth, Development and Pathogenicity ofBotrytiscinerea
摘要
Abstract
[Objective]The objective of this study is to investigate the function ofBcKMO gene in growth, development and pathogenicity ofBotrytis cinerea, to further clarify the molecular mechanism of theBcKMO gene in development and pathogenicity of the fungus and provide a theoretical basis for the gray mould control.[Method] Bioinformatics methods were used to analyze theBcKMO gene and its encoding product. TheBcKMO gene coding region was amplified and cloned into pBARKS1-eGFP, and pBARKS1-BcKMO-eGFP was constructed. The protoplasts of mutant BCG183 were prepared and transformed with pBARKS1-BcKMO-eGFP by PEG-mediated method. PCR, Southern blotting and real-time PCR were used to identify the transformant. Phenotype including colony morphology, mycelium morphology, growth rate, conidial yield, and so on, and pathogenicity assays of wild-type strain BC22 (WT), mutant BCG183 and complemutant BCG183/BcKMO were performed to verify the function of the BcKMO gene in growth, development and pathogenicity.[Result]TheBcKMO gene encodes kynurenine 3-monooxygenase (KMO) having monooxygenase FAD conserved domain and four Aromatic-ring hydroxylase-like motifs. The BcKMO protein displayed high homology to rossmann-fold NAD(P)(+)-binding protein (gi156049701) ofSclerotinia sclerotiorum and FAD-dependent oxidoreductase (gi512192943) ofOphiostoma piceae. The expression vector of theBcKMO gene, pBARKS1-BcKMO-eGFP, was successfully constructed and transformed into the protoplasts of mutant BCG183. Results of PCR, Southern blotting and Real-time PCR showed that theBcKMO gene complementing mutant (BCG183/BcKMO) was successfully obtained. Phenotype assay showed that the mutant BCG183 growed slowly, did not produce conidia and sclerotia, and hyphae slender, but the phenotypes of the BCG183/BcKMO were all similar to that of WT. Compared with WT and the BCG183/BcKMO, the pathogenicity of the mutant BCG183 on kidney bean leaves and cucumber leaves were dramatically increased.[Conclusion]TheBcKMO gene is a negative regulation in growth and development, and a positive regulation in pathogenicity ofB. cinerea.关键词
灰葡萄孢/BcKMO/突变体/生长发育/致病力Key words
Botrytis cinerea/BcKMO/mutants/growth and development/pathogenicity引用本文复制引用
李培芬,董金皋,赵福鑫,董丽萍,郑会欣,赵斌,张靖,司贺龙,邢继红,韩建民..灰葡萄孢BcKMO在病菌生长、发育和致病过程中的功能[J].中国农业科学,2014,(15):2971-2979,9.基金项目
河北省科技支撑计划项目 （）
