Botryosphaeria dothidea突变体库的构建及分析

Construction and Evaluation of the Mutant Library of Botryosphaeria dothidea

Botryosphaeria dothidea是重要的林果溃疡病害病原,分布广,危害重。本研究应用ATMT(Agrobacterium tumefaciens-mediated transformation)介导的B.dothidea遗传转化体系成功构建了1053个转化子的突变体库,并且通过继代培养、PCR验证和Southern blot证明潮霉素B抗性基因整合到B.dothidea基因组中且可稳定遗传。以B.dothidea sdau11-126为对照,对526株转化子的菌落形态、生长速率和致病性进行分析,筛选获得了8个变异明显且稳定的突变体,以期为B.dothidea致病基因的分离、克隆和功能鉴定奠定基础。

Botryosphaeria dothidea is an important pathogen of forest and fruit trees and it is widely distributed and causes serious damage. In this study, mutant library of B. dothidea was constructed via ATMT mediated transformation and 1053 transformants were obtained. Hygromycin B resistance genes were integrated into the genome of the B. dothidea, and mitotically stable after several subcultures were tested by PCR amplification and Southern blot. The strain B. dothidea sdau11-126 was used as control to analyze colonial morphology, growth rate and pathogenicity of 526 transformants. Eight stable and obvious mutants were obtained by screening. This study will lay a foundation for isolating, cloning and functional identification of the pathogenic genes.