文献类型： 外文期刊
作者： Wang, Ya-Jie 1 ; Lu, Xiao-Hua 2 ; Zhen, Xing-Hou 3 ; Yang, Hui 3 ; Che, Yan-Nian 3 ; Hou, Jing-Yi 3 ; Geng, Meng-Ting 3 ; Liu, Jiao 1 ; Hu, Xin-Wen 3 ; Li, Rui-Mei 1 ; Guo, Jian-Chun 1 ; Yao, Yuan 1 ;
作者机构： 1.Chinese Acad Trop Agr Sci, Inst Trop Biosci & Biotechnol, Key Lab Biol & Genet Resources Trop Crops, Minist Agr, Haikou 571101, Hainan, Peoples R China
2.Hainan Inst Trop Agr Resources, Key Lab Biol & Genet Resources Trop Crops Hainan, Haikou 571101, Hainan, Peoples R China
3.Hainan Univ, Sch Life Sci, Haikou 570228, Hainan, Peoples R China
4.Chinese Acad Trop Agr Sci, San Yan Res Inst, Sanya 572025, Peoples R China
5.Hainan Yazhou Bay Seed Lab, Sanya 572025, Peoples R China
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
页码：
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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