摘要
以紫茎泽兰为受体材料,GUS基因为报告基因,对农杆菌介导的遗传转化条件及影响因素进行研究,建立了农杆菌介导的紫茎泽兰遗传转化体系。结果表明,将未经过预培养的幼叶外植体在OD600为0.4的稀释菌液中浸泡12min,共培养3d后,转移到加有卡那霉素50mg/L(筛选压)和羧苄青霉素250mg/L的分化培养基上,经过20d外植体直接分化出不定芽,诱导生根成苗。经PCR和组织化学染色鉴定,可稳定获得较高的阳性转化率。
With Eupatorium adenophorum Sprcng as acceptor materials,GUS gene as reporter gene,a high efficient Agrobacterium tumefaciens-mediated genetic transformation of E. adenophorum was established. Meanwhile,relevant conditions and factors were studied. The final optimal procedure includes dipping the explants which were not pre-trained in the diluted Agrobacterium tumefaciens liquid （ OD600 = 0.4） for 12 min, after co-culturing for 3 days, and transfering the explants to differentiation medium containing 50 mg/L kanamycin（ as selecting pressure）and 250 mg/L carbencillin. The adventicious shoots was induced after about 20 days,then induction of roots. Potsitive transgenctic plants were obtained by PCR and histochemical staining analysis.
出处
《生物技术通报》
CAS
CSCD
北大核心
2010年第7期153-156,共4页
Biotechnology Bulletin
基金
农业部公益性行业科研专项(200803021-030)
关键词
紫茎泽兰
农杆菌
遗传转化
GUS基因
Eupatorium adenophorum Spreng Agrobacterium tumefaciens Genetic transformation GUS gene
