Chinese Bulletin of Botany ›› 2021, Vol. 56 ›› Issue (1): 62-70.DOI: 10.11983/CBB20174

• TECHNIQUES AND METHODS • Previous Articles     Next Articles

Establishment of Biolistic Mediated Transformation System for Elymus sibiricus

Pengfei Du1,2,3, Yu Wang2, Yingping Cao2, Song Yang2, Zhichao Sun2, Decai Mao3,4, Jiajun Yan3,4, Daxu Li3,4, Meizhen Sun5, Chunxiang Fu2,*(), Shiqie Bai3,4,*()   

  1. 1Southwest University for Nationalities, Chengdu 610041, China
    2Shandong 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
    3Sichuan Grassland Science Research Institute, Chengdu 611731, China
    4Engineering Research Center for Ecological Restoration of Alpine Grassland on the Qinghai-Tibet Plateau, National Forestry and Grassland Administration, Chengdu 611731, China
    5Qingdao Blood Center, Qingdao 266071, China
  • Received:2020-11-02 Accepted:2021-01-05 Online:2021-01-01 Published:2021-01-15
  • Contact: Chunxiang Fu,Shiqie Bai

Abstract: 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.

Key words: Elymus sibiricus cv. ‘Chuancao No.2’, inflorescence, Agrobacterium infection, biolistic mediated transformation, callus treatment