灰葡萄孢T-DNA插入细胞壁缺陷突变体的筛选  

Isolation of the cell wall defect mutants by T-DNA insertion in Botrytis cinerea

作　　者：雷娜[1] 张为宏[1] 朱廷恒[1] 汪琨[1] 崔志峰[1] 

机构地区：[1]浙江工业大学生物与环境工程学院,浙江杭州310032

出　　处：《浙江农业学报》2011年第4期754-758,共5页Acta Agriculturae Zhejiangensis

基　　金：浙江省自然科学基金资助项目(Y307492)

摘　　要：用荧光增白剂Calcofluor White(CFW)对234株T-DNA插入灰葡萄孢突变株进行筛选,获得了3株对CFW敏感性(B-117,B-169,D-9)和1株对CFW抗性(B-62)的突变株。1.2 mol.L-1Sorbito对D-9的生长缓慢有挽救作用。在SDS培养基上D-9和野生型的差异不大,但在NaCl培养基上野生型的生长受到抑制,D-9则不受NaCl影响。在孢子萌发试验中,D-9突变株也与野生型明显不同,表现为D-9的孢子膨大、菌丝较粗、长度增加而且不弯曲。在番茄感染试验中,D-9突变株侵染番茄的毒力大幅度减弱。由此推测D-9突变株与细胞壁缺损以及致病性有关。234 T-DNA insertion mutants of Botrytis cinerea were screened by using Calcofluor White,and three sensitive mutants（B-117,B-169,D-9）and one resistant mutant（B-62）were obtained.In the test with isotonic PDA plates containing 1.2 mol·L^-1 sorbitol,the growth of D-9 was not affected or rescued,while the growth of wild type was suppressed.Also,the growth of wild type was suppressed by 0.6 mol·L-1 NaCl but D-9 was not affected.No obvious difference was found between wild type and D-9 in the SDS medium.The spores of D-9 appeared swollen,its hyphae were thicker,and the length increased without bending,which was obviously different from that of wild type in the spore germination assay.In the experiment of tomato infection,the virulence of D-9 mutant was significantly reduced,almost no infection phenomenon can be observed in the period of six days.These results indicated that D-9 mutant was probably related to cell wall defect and pathogenicity.

关 键 词：灰葡萄孢 T-DNA插入突变体 CFW敏感性 细胞壁缺陷 

分 类 号：Q933[生物学—微生物学]