摘要
甘蔗梢腐病是甘蔗生长中期的主要真菌病害。目前,研究甘蔗梢腐病病原菌和甘蔗品种的互作机制是防治梢腐病害的重点。本研究运用农杆菌介导的转化方法将含有绿色荧光蛋白(GFP)的双元载体PCAMBgfp转化到甘蔗梢腐病野生菌株(YN41)中,获得41个转化子。随机挑取7个转化子进行PCR扩增,均可扩增出潮霉素基因目标条带;且转化菌株的菌落形态、生长速度和致病力与野生型菌株相比没有显著差异;单孢继代培养后仍能发出稳定的绿色荧光,并且能够稳定遗传。同时发现60μg/m L潮霉素B能够完全抑制甘蔗梢腐病菌株YN41的生长。以上结果表明GFP基因已成功转化到甘蔗梢腐病菌株YN41中,可为后续甘蔗梢腐病病原菌和甘蔗品种的互作机制研究提供技术支撑。
Pokkah boeng disease is an devastating fungal diseases at the elongation stage of sugarcane. Analysis of the interaction mechanism between Fusarium moniliformae and sugarcane variety is the key point for curing this disease. In this research, total 41 transformants were got in F. moniliformae by expression of Agrobacterium tumefaciens containing p CAMBgfp（p CAMB： gfp： hyg）, in which seven clones were randomly selected and verified by PCR amplifying with the hyg primers. The mycelia and conidia of these positive clones was similar with the wild-type strain YN41 under fluorescence microscopy. Through pathogenicity and genetic stability test, these positive clones also show the same as YN41. And 60 μg/m L hygromycin B could completely inhibit F. moniliformae YN41 growth. All these results showed that GFP had been successfully transformed into the F. moniliformae YN41, which will provide powerful tool for the further study of the interaction mechanism between F. moniliformae and sugarcane varieties.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2016年第5期1189-1194,共6页
Genomics and Applied Biology
基金
地区科学基金项目(甘蔗梢腐病病原菌的致病性分化及其种属特异(LS)染色体区的基因鉴定31460374)
亚热带农业生物资源保护与利用国家重点实验室基础性专项研究课题(甘蔗梢腐病病原菌的基因组分析与致病基因的鉴定)共同资助
关键词
甘蔗梢腐病病原菌
绿色荧光蛋白(GFP)
农杆菌
遗传转化
Sugarcane pokkah boeng pathogen
Green fluorescent protein（GFP）
Agrobacterium tumefaciens
Genetic transformation
