摘要
利用发根土壤杆菌农杆碱型15834、A_4菌株和甘露碱型8196、K_(599)菌株体外转化甘草子叶及下胚轴。结果表明:除K_(599)菌株外,其余三种菌株都能诱导产生甘草发状根无性系。其中15834菌株的诱导率最高,下胚轴的诱导率高于子叶。组织学观察表明,在感染后4天左右,下胚轴维管束鞘外细胞、特别是形成层部分细胞大量启动形成分生细胞及分生细胞团,并进一步分裂形成根原基。6—8天后根原基具单向极性生长形成完整的发状根结构。发状根能在无外源激素的培养基上迅速生长。高压纸电泳检测到发状根中存在农杆碱及甘露碱,说明Ri质粒T-DNA编码的这两种冠瘿碱合成酶基因在甘草细胞中得到了表达。
Agrobacterium rhizogenes agropine type strains 15834, A_4 and mannopine type strains 8196, K599 are used to transform cotyledons and hypocotyls of Glycyrrhiza uvalensis. The results show that all the other strains induce hairy root clones except K599. The transformation efficiency of strain 15834 is the highest among the three strains, and the hypocotyl transformation efficiency is higher than the cotyledon's. Histological observation show that four days after infection, the cells outside vascular bundle sheath, especially the cambium cells redifferentiate to form root primordia, and the intact hairy roots form after about 8 days. The hairy root grow extensively on hormone-free medium. High voltage paper electrophoresis detected agropine and mannopine in the hairy roots. The results demonstrate that the opine synthase gene coded by Ri T-DNA has transferred, integrated and expressed in transformed tissues.
出处
《武汉植物学研究》
CSCD
1991年第4期301-305,共5页
Journal of Wuhan Botanical Research
关键词
甘草
发根土壤杆菌
子叶
下胚轴
Glycyrrhiza uralensis Fisch
Agrobacterium rhizogenes
Hairy root
Histology
