A Transformation and Genome Editing System for Cassava Cultivar SC8

文献类型： 外文期刊

第一作者： Wang, Ya-Jie

作者： Wang, Ya-Jie;Liu, Jiao;Li, Rui-Mei;Guo, Jian-Chun;Yao, Yuan;Wang, Ya-Jie;Lu, Xiao-Hua;Liu, Jiao;Li, Rui-Mei;Lu, Xiao-Hua;Zhen, Xing-Hou;Yang, Hui;Che, Yan-Nian;Hou, Jing-Yi;Geng, Meng-Ting;Hu, Xin-Wen;Zhen, Xing-Hou;Yao, Yuan;Zhen, Xing-Hou;Yao, Yuan

作者机构：

关键词： cassava; CRISPR/Cas9; efficient transformation; friable embryogenic calli; homozygous; SC8

期刊名称：GENES （ 影响因子：4.141； 五年影响因子：4.474 ）

ISSN：

年卷期： 2022 年 13 卷 9 期

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收录情况： SCI

摘要： Cassava starch is a widely used raw material for industrial production. South Chinese cassava cultivar 8 (Manihot esculenta Crantz cv. SC8) is one of the main locally planted cultivars. In this study, an efficient transformation system for cassava SC8 mediated with Agrobacterium strain LBA4404 was presented for the first time. Cassava friable embryogenic calli (FECs) were transformed through the binary vector pCAMBIA1304 harboring GUS- and GFP-fused genes driven by the CaMV35S promoter. The transformation efficiency was increased in the conditions of Agrobacterium strain cell infection density (OD600 = 0.65), 250 mu M acetosyringone induction, and agro-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 MePDS gene with a YAO promoter-driven CRISPR/Cas9 system were generated. This study will open a more functional avenue for the genetic improvement of cassava SC8.

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