溶杆菌SNNU513基因gfp标记及在玉米根部定殖  

Colonization of gfp-tagged Lysobacter sp.SNNU513 on Maize's Root

作　　者：武坤毅 王斐斐[1] 崔浪军[1] 章华伟[2] 白成科[1] 

机构地区：[1]药用植物资源与天然药物化学教育部重点实验室/陕西师范大学生命科学学院,西安710062 [2]浙江工业大学药学院,杭州310014

出　　处：《中国生物防治学报》2014年第1期134-142,共9页Chinese Journal of Biological Control

基　　金：国家自然科学基金(31101481);浙江省公益性技术应用研究项目(2012C32010)

摘　　要：溶杆菌属细菌在植物病害生物防治中有着广阔的应用潜力。本文以本实验室从中药材远志分离筛选的溶杆菌属新菌株Lysobacter sp.SNNU513为材料,筛选出高效制备该菌株感受态细胞Inoue法,电击转化条件为场强20 kV/cm、电脉冲时间5 ms时可将含绿色荧光蛋白基因gfp的质粒pGLO导入该菌株感受态细胞中,重组菌SNNU513-pGLO能高效、稳定表达绿色荧光蛋白基因,与出发菌株的生长特性、抑菌活性等生物学特性差异不显著。将成功构建的重组菌SNNU513-pGLO用于检测该菌株在玉米根部的定殖规律,结果表明,定殖量从表皮到韧皮部有明显减少趋势。Lysobacter sp. SNNU513 isolated from Radix polygalae rhizospher could inhibit pathogenic fungus. In this study, the preparation of the efficient competent cells were screened out and pGLO plasmid was introduced into the competent cells by electroporation and transformation. The results showed that the recombinant strain SNNU513-pGLO could efficiently and stably express the green fluorescent protein gene （gfp） when the electric field strength was 20 kV/cm and the electric pulse time was 5 ms. The percent of the green cells was 100%when the strain SNNU513-pGLO was recirculated in the non-selective medium for 20 times. Both strains SNNU513-pGLO and SNNU513 showed the same growth characteristics and the same inhibition of Rhizotonia cerealis in vivo. The gene gfp was transferred into original strain Lysobacter sp. SNNU513 by electroporation successfully. The gfp-tagged strain adhering more tightly to the velamen than the phloem could colonize on the surface and interior of maize’s root.

关 键 词：生防菌 溶杆菌属 生物学特性 定殖 

分 类 号：S476[农业科学—农业昆虫与害虫防治]