A Transformation and Genome Editing System for Cassava Cultivar SC8

• Published in: Genes Vol. 13; no. 9; p. 1650
• Main Authors: Wang, Ya-Jie, Lu, Xiao-Hua, Zhen, Xing-Hou, Yang, Hui, Che, Yan-Nian, Hou, Jing-Yi, Geng, Meng-Ting, Liu, Jiao, Hu, Xin-Wen, Li, Rui-Mei, Guo, Jian-Chun, Yao, Yuan
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.09.2022; MDPI
• Subjects: Acetosyringone; Agrobacterium; Antibiotics; Cassava; Cell size; CRISPR; CRISPR/Cas9; Crop diseases; Cultivars; Efficiency; efficient transformation; Embryos; Equipment and supplies; friable embryogenic calli; Gene Editing; Genes; Genetic aspects; Genetic engineering; Genetic transformation; Genomes; homozygous; Manihot - genetics; Manihot esculenta; Mutation; Protocol; SC8; Starch - metabolism; Transformation, Genetic; China; Africa; homozygous; SC8; efficient transformation; CRISPR/Cas9; cassava; friable embryogenic calli
• ISSN: 2073-4425; 2073-4425
• DOI: 10.3390/genes13091650

Cassava starch is a widely used raw material for industrial production. South Chinese cassava cultivar 8 ( Crantz cv. SC8) is one of the main locally planted cultivars. In this study, an efficient transformation system for cassava SC8 mediated with strain LBA4404 was presented for the first time. Cassava friable embryogenic calli (FECs) were transformed through the binary vector pCAMBIA1304 harboring and fused genes driven by the promoter. The transformation efficiency was increased in the conditions of strain cell infection density (OD = 0.65), 250 µM acetosyringone induction, and -cultivation with wet FECs for 3 days in dark. Based on the optimized transformation protocol, approximately 120-140 independent transgenic lines per mL settled cell volume (SCV) of FECs were created by gene transformation in approximately 5 months, and 45.83% homozygous mono-allelic mutations of the gene with a promoter-driven CRISPR/Cas9 system were generated. This study will open a more functional avenue for the genetic improvement of cassava SC8.