沟叶结缕草遗传转化体系的建立  

Establishment of Genetic Transformation System for Zoysia matrella

作　　者：王凯 汪毅[1] 曲爱爱 王蕊 郭海林[1] 李晓慧 佘建明[1] 宗俊勤[1] 李建建[1] 刘建秀[1] WANG Kai;WANG Yi;QU Aiai;WANG Rui;GUO Hailin;LI Xiaohui;SHE Jianming;ZONG Junqin;LI Jianjian;LIU Jianxiu(Institute of Botany,Jiangsu Province and Chinese Academy of Sciences(Nanjing Botanical Garden Mem.Sun Yat-Sen),Nanjing,Jiangsu 210014,China)

机构地区：[1]江苏省中国科学院植物研究所(南京中山植物园),江苏南京210014

出　　处：《热带作物学报》2020年第8期1566-1573,共8页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金项目(No.31672195,No.31572155)。

摘　　要：为构建有效的沟叶结缕草遗传转化体系,以沟叶结缕草愈伤组织为受体材料,通过农杆菌介导法将笔者单位克隆的耐盐相关基因ZmPDI基因转入野生型植株中,研究共培养时间、侵染液浓度、侵染时间和抗生素浓度等因子对转化效率的影响。结果表明:最佳遗传转化体系为菌株OD600值为0.4,侵染30 min,共培养3 d后进行选择培养。特美汀在选择培养阶段最适抑菌浓度为250 mg/L,潮霉素愈伤组织筛选的最适选择压力为40 mg/L,苗筛的最适浓度为15 mg/L。通过GUS活性的组织化学分析和PCR鉴定,显示目的基因已成功转入沟叶结缕草基因组中。Zoysia matrella is a perennial warm-season turfgrass,one of halophytes with the salt-tolerantce,and is widely used in temperate zone and subtropical regions.However,its complete genetic transformation system have not been reported.In this study,the callus from Z.matrella was used as receptor material and the genetic transformation of the salt-tolerant gene ZmPDI was carried out by Agrobacterium-mediated method to study a series of conditions for transformation efficiency including the co-culture time,concentration of the infection,and infection time.The results showed that the optimal genetic transformation was supposed to be:agrobacterium concentration OD600=0.4,infected for 30 minutes,co-cultured for 3 days and then followed by selective culture.During the whole process,the optimum concentration of timentin in the postponed culture medium was 250 mg/L.As for hygromycin,the optimal selection pressure for callus was 40 mg/L,while the best concentration for rooting screening was 15 mg/L.The histochemical analysis ofβ-glucuronidase(GUS)activities and PCR identification indicated that the ZmPDI gene was transferred into the genome by hygromycin resistance selection.

关 键 词：沟叶结缕草 蛋白二硫键异构酶(PDI) 根癌农杆菌 遗传转化 

分 类 号：S688.4[农业科学—观赏园艺]