根癌农杆菌介导转化川草二号老芒麦胚性愈伤组织

以川草二号老芒麦成熟种子为外植体，经过对培养基的筛选和培养条件优化，建立了愈伤组织再生系统。转化载体为pCAMBIA1304质粒，其T—DNA上携有潮霉素抗性基因（hptII）和类产碱假单胞菌杀虫蛋白基因（ppIP），经根癌农杆菌EHA105介导转化结构致窑、颗粒状、黄白色的胚性愈伤组织。通过潮霉素筛选和对抗性植株进行分子检测，获得了转基因植株。同时优化了农杆菌遗传基因转化的参数，建立了农杆菌介导的川草二号老芒麦程序化转基因方案。

Transformation of Embryogenic Calli of Siberian Wildrye Grass (Elymus sibiricus L. cv. Chuancao No.2) Mediated by Agrobacterium

Formation of embryogenic calli of Si-berian wildrye grass (Elymus sibiricus L. cv.Chuancao No.2) was induced from mature seedsas explants, and proliferated on MS medium con-taining 2,4-D 5.0 mg/L and KT 0.05 mg/L. An ef-fective and stable callus regeneration system wasestablished by optimizing the culture conditions(Tables 1, 2 and Fig.2). After the calli were sub-cultured 8 weeks, selected the whitish-yellow-coloured compact nodular calli that transformedwith plasmid pCAMBIA1304 carrying hygromycinresistance gene (hptII) and Pseudomonas pseudoal-caligenes insecticidal protein gene (ppIP), whichwas mediated by an Agrobacterium strain EHA105.Resistant plants were obtained after hygromycinselection (Figs.3, 4). Some important factors thataffect the transformation efficiency were studied,which included selection pressure, time of embryo-genic calli proliferation, OD value of Agrobacte-rium suspension, temperature, medium and timeof co-cultivation, and concentration of antibioticsused for suppressing the overgrowth ofAgrobacterium in the course of transformationplant regeneration. This research is the first suc-cessful genetic transformation of Elymus sibiricusL. cv. Chuancao No.2 mediated by Agrobacterium.