灰葡萄孢T-DNA插入细胞壁缺陷突变体的筛选

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-1 Sorbito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。

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

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.