生防链霉菌SSD49的绿色荧光蛋白标记及其在毛白杨组培苗中的定殖

Green Fluorescent Protein Marker of Biocontrol Streptomyces SSD49 and Its Colonization on the Populus tomentosa Somaclone

杨树溃疡病是中国杨树人工林重大生物灾害之一,采用生物防治的方法控制杨树溃疡病是持续有效的手段。为了获取能够高效定殖并对杨树溃疡病菌有良好生防效果的拮抗菌株,通过引入强启动子(erm Ep)构建了高表达绿色荧光蛋白(Green fluorescent protein,GFP)的重组质粒,进而通过接合实验将该质粒导入到链霉菌生防菌株SSD49,构建了绿色荧光蛋白标记菌株SSD49-p IJ8660Ep,利用荧光显微镜研究该生防菌在毛白杨组培苗中的定殖情况。结果表明,SSD49标记绿色荧光蛋白后没有影响其对杨树溃疡病病原菌的抑菌活性,标记菌株能够定殖于毛白杨组培苗的茎和叶中。成功将杨树溃疡病生防菌株SSD49进行了绿色荧光蛋白标记,并且该菌株在毛白杨组培苗中有一定的定殖能力。

Poplar canker is one of the major biohazard of poplar plantation in China, and the sustained and effective method to control poplar canker is biological c, ontrol. This study aims to obtain a strain which has solid bio-control effect on Botryosphaeria dothidea and can colonize in poplar efficiently. This study imported a strong promoter （ ermEp ） and constructed highly-expressed green fluorescent protein （ GFP ） recombinant plasmid, then led the plasmid into bio-control strain Streptomyces SSD49 through joint experiment, and constructed the GFP-tagged strain SSD49-pIJ8660Ep. Fluorescence microscope was used to study the colonization of this biocontrol strain in Populus tomentosa somaclone. Results showed that the GFP-tagged SSD49 didn＇ t affect its bacteriostatic activity against poplar canker pathogen and colonized in stems and leaves of P, tomentosa somaclone. This study successfillly constructed the GFP-tagged strain SSD49, and the strain presented certain colonization in P. tomentosa somaclone.