摘要
以大花蕙兰原球茎(PLBs)为外植体,采用EHA105和LBA4404 2种根癌农杆菌菌株与pCAMBIA1301质粒构建工程菌介导,以建立大花蕙兰遗传转化体系,并比较不同受体处理方式、菌液浓度和侵染方式等对大花蕙兰转化的影响.结果表明:(1)以切成3 mm左右的PLBs小块作为受体材料,用OD600值为0.6的LBA4404根癌农杆菌菌株,并用MS+1.0 mg/L BA+200μmol/L AS(乙酰丁香酮)的液体培养基将菌液等体积稀释侵染,转化率可达62.5%.(2)大花蕙兰对潮霉素(Hyg)十分敏感,5 mg/L Hyg对转化后的PLBs有较好的筛选效果,筛选后最高成活率为13.0%.(3)PCR检测初步证明,通过根癌农杆菌介导的方法获得了2株转基因大花蕙兰植株.
A transformation system of Cymbidium hybridum PLBs（protocorm-like bodies） was established with Agrobacterium tumefaciens strains EHA105 and LBA4404 that harbored binary plasmid vector pCAMBIA 1301.The effects of different receptor treatments,the bacterial concentrations and infection methods on the transformation efficiency of C.hybridum PLBs were compared.The results showed that efficient transformation could be gotten with 3 mm PLBs blocks as receptor.Higher transformation rate was gained when the OD600 of LBA4404 was about 0.6 and bacteria was diluted with equivalent volume of MS＋1.0 mg/L 6-BA＋200 μmol/L AS liquid medium.The highest transformation rate was 62.5%.C.hybridum was very sensitive to hygromycin（Hyg）.The better selective survival rate of 13.0% was gotten when the concentration of Hyg was 5 mg/L.Successfully transformed C.hybridum PLBs were confirmed by PCR analysis.
出处
《西北植物学报》
CAS
CSCD
北大核心
2011年第1期27-32,共6页
Acta Botanica Boreali-Occidentalia Sinica
基金
教育部留学回国人员科研启动基金
关键词
大花蕙兰
根癌农杆菌
遗传转化
类原球茎
Cymbidium hybridum
Agrobacterium tumefaciens
genetic transformation
protocorm-like bodies（PLBs）
