河南农业科学2012，Vol.41Issue(1)：82-86,5.
农杆菌介导尖孢镰刀菌西瓜专化型转化体系的建立及突变体筛选
Agrobacterium tume faciens-mediated Transformation of Fusarium oxysporum f.sp.niveumis and Screening of Mutants with Special Traits
摘要
Abstract
In this study, Fusarium oxysporurn f. Sp. Niveumis FO-11-06 was successfully transformed mediated by Agrobacterium tumefaciens, and T-DNA insertion mutants were further screened. The effects of pH and cultivation media on genetic transformation were studied under the condition of 25 ℃ , 1 X 106 spores/mL,A. Tumefaciens (AGL1) OD600 =0. 15,200 /μmol/L of acetosyringone and 48 h of co-cultivation in the presence of induction medium. The results showed that the transformants could be obtained in specific pH value range from 5. 1 to 5. 8 and the transformation efficiency reached the highest level at pH 5. 3, with about 553 transformants per 106 spores. It also revealed that transformation efficiency was higher on cellophane (551 transformants per 106 spores) and NC millipore filter (549 transformants per 106 spores) compared to filter paper. Hygromycin resistance and PCR assay of the transformants showed that the T-DNA had been successfully integrated into the genome of F. Oxysporum f. Sp. Niveumis FO-11-06. Forty six mutants with specific traits were identified from 1 989 transformants by assay of colony growth, colony color,and conidia output. The transformation system and mutants provided a basis for the study of development,pathogenicity mechanism and functional gene of the fungus.关键词
尖孢镰刀菌西瓜专化型/农杆菌/T-DNA插入转化/转化效率Key words
Fusarium oxysporum f. Sp. Niveumis/Agrobacterium tumefaciens /T-DNA transformation/transformation efficiency分类
农业科技引用本文复制引用
梁慎,薛保国,常高正,赵卫星,杨丽荣,孙虎,李晓慧,杨帆,全鑫,徐小利..农杆菌介导尖孢镰刀菌西瓜专化型转化体系的建立及突变体筛选[J].河南农业科学,2012,41(1):82-86,5.基金项目
国家西甜瓜产业技术体系专项经费资助项目(CARS-26-33) （CARS-26-33）
