摘要
辣椒溶杆菌(Lysobacter capsici)X2-3是本实验室从小麦根际土壤中分离的对多种植物病原真菌和卵菌有明显拮抗活性的菌株,建立有效的遗传操作体系对了解其基因的功能具有重要意义。本研究以VgrG为目标基因,比较了不同载体对X2-3基因敲除效果,结果发现,载体pEX18GM、pBR325和pK19mob转化X2-3后,均未得到VgrG缺失突变体;而载体pKMS1转化X2-3后,经10%蔗糖二次筛选、PCR和Southern blot验证等,成功得到VgrG缺失突变体;用广宿主载体pBBR1MCS-5构建的VgrG互补突变体,可以恢复VgrG基因的功能。本结果为进一步研究该菌的基因功能提供了技术保障。
Lysobacter capsici X2-3 was isolated by our lab from wheat rhizosphere soil with antimycotic activities to many plant pathogenic fungi and oomycetes. Establishing an effective genetic manipulation system was of great significance for understanding the function of its genes. Using VgrG gene as target, the study compared gene knockout effect of four different vectors on X2-3. The results showed that X2-3 transformed by the vectors pEX18GM, pBR325 and pK19mob were not VgrG deletion mutates. However, after transformation of vector pKMS1 into X2-3, VgrG deletion mutants were successfully obtained by 10% sucrose secondary screening, PCR and Southern blot verification. The complemented mutants of VgrG constructed by broad-host vector pBBR1MCS-5 also restored the function of VgrG gene. This study provided a technical guarantee for the further study on the gene function of the strain X2-3.
作者
王虹
李星志
赵丹
刘爱新
Wang Hong;Li Xingzhi;Zhao Dan;Liu Aixin(College of Plant Protection,Shandong Agricultural University,Taian 271018,China)
出处
《山东农业科学》
2019年第4期7-12,共6页
Shandong Agricultural Sciences
基金
国家重点研发计划项目"重要作物病害生防微生态制剂的研制与应用"(2017YFD0201106-11)
山东省科技发展计划项目(2013GNC11015)
