摘要
利用发根农杆菌A4菌株侵染三分三组培苗叶片诱导毛状根,并对其毛状根进行继代增殖培养;采用PCR技术检测毛状根。结果表明:发根农杆菌A4菌株Ri质粒的rolB基因已整合到三分三叶肉细胞的基因组中,进而成功地从叶片诱导出毛状根,诱导率可达77%以上;毛状根除菌后,能在无激素的MS固体培养基上快速生长。三分三遗传转化体系的建立为实现基于毛状根的三分三产托品烷类生物碱的代谢工程奠定基础。
Agrobacterium rhizogenes A4 was used to induce hairy roots by infecting the leaves of Anisodus acutangulus,the hairy roots were subcultured and multiplied in the current investigation.The PCR technique was applied to detect hairy roots.The results showed that rolB gene in Ri plasmid of Agrobacterium rhizogenes A4 was easily integrated into the genome of mesophyll cell of Anisodus acutangulus,and the hairy roots were induced successfully from the leaves at the frequency of more than 77%.The hairy roots that eliminated bacteria could grow rapidly on MS solid medium with hormone-free.The establishment of genetic transformation systems in Anisodus acutangulus may provide a basement for the metabolic bioengineering of tropine alkaloids via hairy root pathway.
出处
《山地农业生物学报》
2011年第2期115-119,共5页
Journal of Mountain Agriculture and Biology
基金
贵州省中药材现代产业技术体系建设项目(GZCYTX2010-0201)
关键词
三分三
发根农杆菌A4菌株
遗传转化
毛状根
Anisodus acutangulus
Agrobacterium rhizogenes A4
Genetic transformation
Hairy roots
