基因枪介导的老芒麦遗传转化体系的建立  

Establishment of Biolistic Mediated Transformation System for Elymus sibiricus

作　　者：杜鹏飞 王玉 曹英萍[2] 杨松 孙志超 毛德才[3,4] 鄢家俊 李达旭[3,4] 孙美贞 付春祥[2] 白史且 Pengfei Du;Yu Wang;Yingping Cao;Song Yang;Zhichao Sun;Decai Mao;Jiajun Yan;Daxu Li;Meizhen Sun;Chunxiang Fu;Shiqie Bai(Southwest University for Nationalities,Chengdu 610041,China;Shandong Technology Innovation Center of Synthetic Biology,Shandong Provincial Key Laboratory of Energy Genetics,Key Laboratory of Biofuels,Qingdao Institute of Bioenergy and Bioprocess Technology,Chinese Academy of Sciences,Qingdao 266101,China;Sichuan Grassland Science Research Institute,Chengdu 611731,China;Engineering Research Center for Ecological Restoration of Alpine Grassland on the Qinghai-Tibet Plateau,National Forestry and Grassland Administration,Chengdu 611731,China;Qingdao Blood Center,Qingdao 266071,China)

机构地区：[1]西南民族大学,成都610041 [2]中国科学院青岛生物能源与过程研究所,中国科学院生物燃料重点实验室,山东省能源生物遗传资源重点实验室,山东省合成生物技术创新中心,青岛266101 [3]四川省草原科学研究院,成都611731 [4]国家林草局青藏高原高寒草地生态修复工程技术研究中心,成都611731 [5]青岛市中心血站,青岛266071

出　　处：《植物学报》2021年第1期62-70,共9页Chinese Bulletin of Botany

基　　金：首批万人计划四川天府杰出科学家项目(2018);四川省重点研发项目(No.2019YFN0170);西南民族大学研究生创新型科研项目(No.CX2019SZ91);山东省农业良种工程(No.2019LZGC010)。

摘　　要：川草2号老芒麦(Elymus sibiricus)是青藏高原地区治理荒漠化和建设高产人工草地的主要栽培草种。用川草2号老芒麦5种外植体诱导愈伤组织,经分化测试,仅幼穗愈伤组织能分化再生。以当代培养25天和35天的结构致密坚硬的幼穗愈伤组织为受体,分别进行农杆菌侵染和基因枪转化,结果只有基因枪能转化成功。在基因枪转化过程中,采用高渗培养和滤纸干燥2种方式预处理愈伤组织,结果表明滤纸干燥处理比高渗处理转化效率高。当代诱导25天的幼穗愈伤组织,滤纸干燥处理2小时转化效率最高,达40%。该研究成功获得了基因枪转化的以川草2号老芒麦幼穗愈伤为受体的阳性愈伤组织。Elymus sibiricus cv.’Chuancao No.2’is the main cultivated grass species for desertification control and construction of high-yield and high-quality pasture in northwest Sichuan Plateau.In this study,we tested five explants of E.sibiricus cv.’Chuancao No.2’for callus induction,and found that only inflorescence calli were able to differentiate and regenerate.The calli of inflorescence with dense and hard structure cultured for 25 d and 35 d were used for Agrobacterium and biolistic mediated transformation respectively.The results showed that only biolistic-mediated transformation could produce positive transgenic calli of’Chuancao No.2’.In the process of biolistic-mediated transformation,the calli was pretreated in two ways:hyperosmotic culture and filter paper drying.The results revealed that the transformation efficiency of filter paper drying was higher than that of hyperosmotic treatment.For the inflorescence callus after 25 d induction,the transformation efficiency under the condition of 2 h drying of filter paper was highest which reached about 40%.In short,we applied the biolistic technology in’Chuancao No.2’for the first time and successfully obtained the positive transgenic inflorescence calli.This work will lead to establishment of the robust transformation system for E.sibiricus in future.

关 键 词：川草2号老芒麦 幼穗 农杆菌侵染 基因枪转化 愈伤组织处理 

分 类 号：S543.9[农业科学—作物学]