摘要
为获得灰葡萄孢的致病相关基因并研究其基因功能,筛选灰葡萄孢的T-DNA插入突变体库,获得了一株致病力增强的突变体BCt98。利用PCR和Southern Blotting技术,对突变体BCt98进行鉴定。利用TAIL-PCR技术结合生物信息学方法,确定了突变体BCt98中T-DNA插入位点位于BC1G_07014.1基因的第3个外显子上。利用RT-PCR技术,确定了突变体BCt98的突变基因为BC1G_07014.1。突变体BCt98生长速度较快,菌落颜色较浅,菌丝较为致密,不产生分生孢子和菌核,且胞壁降解酶(PMG、PG和Cx)及毒素活性较野生型明显增强。表明BC1G_07014.1基因在灰葡萄孢生长、发育和致病力调控方面发挥重要作用,且参与调控病菌的胞壁降解酶活性和毒素活性。
The objective of this study was to obtain pathogenicity-related genes of Botrytis cinerea and to investigate the function of pathogenicity-related genes. An enhanced pathogenicity mutant,named BCt98,was found by screening T-DNA insertional mutant library of Botrytis cinerea and it was testified by PCR and Southern Blotting techniques. T-DNA insertion site was defined in the third exon of BC1G_07014. 1 gene by using TAIL-PCR and bioinformatics methods. The mutant gene was identified as BC1G_07014. 1 by RT-PCR technology. Compared to the wild type strain,the mutant BCt98 growed quickly,colony was white,did not produce conidium and sclerotia,but showed stronger on cell wall degrading enzyme activity and toxin activity. These results showed that the BC1G_07014. 1 gene was involved in growth,development,pathogenicity and involved in regulating cell wall degradation enzyme activity and toxin activity in B. cinerea.
出处
《华北农学报》
CSCD
北大核心
2015年第3期37-41,共5页
Acta Agriculturae Boreali-Sinica
基金
河北省科技支撑计划项目(12226507)
