Chinese Bulletin of Botany ›› 2023, Vol. 58 ›› Issue (2): 298-307.DOI: 10.11983/CBB22240

• TECHNIQUES AND METHODS • Previous Articles     Next Articles

Highly Efficient Gene Editing of Lignin Gene F5H in Switchgrass

Rui Qiu1,2, Feng He2, Rui Li2, Yamei Wang2, Sinian Xing2, Yingping Cao2, Yefei Liu1,2, Xinyue Zhou1, Yan Zhao1,*(), Chunxiang Fu2,*()   

  1. 1Key Laboratory of Grassland Resources of Ministry of Education/Key Laboratory of Forage Cultivation, Processing and Efficient Utilization of Ministry of Agriculture and Rural Areas/College of Grassland, Resources and Environment of Inner Mongolia Agricultural University, Hohhot 010018, China
    2Qingdao New Energy Shandong Laboratory, Shandong Key Laboratory of Energy Biogenetic Resources, Shandong Energy Institute, Key Laboratory of Biofuels, Chinese Academy of Sciences, Qingdao Institute of Bioenergy and Process, Chinese Academy of Sciences, Qingdao 266101, China
  • Received:2022-10-13 Accepted:2023-01-16 Online:2023-03-01 Published:2023-03-15
  • Contact: *E-mail:;

Abstract: Switchgrass (Panicum virgatum) is an important C4 perennial lignocellulosic bioenergy and forage crop. In order to rapidly develop novel germplasm of switchgrass with high cell wall conversion rate, we cloned the ferulic acid 5-hydroxylase gene (PvF5H) from the heterotetraploid switchgrass cultivar Alamo. According to the PvF5H sequence, the editing target sgRNA was designed and used to construct CRISPR/Cas9-PvF5H editing vector. Finally, 59 positive transgenic switchgrass lines were generated by Agrobacterium tumefaciens mediated transformation. Sequencing analysis showed that the PvF5H was edited in most of the transgenic lines (94.9%), and the homozygous editing efficiency was 55.4%. Thus, we have successfully established a highly efficient gene editing system for switchgrass. It will facilitate manipulating target genes involved in cell wall quality and can be employed to breed novel switchgrass cultivars for production of biofuels and fodders in future.

Key words: bioenergy and forage grass, switchgrass, lignin, F5H, gene editing