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Development of cassava common mosaic virus-based vector for protein expression and gene editing in cassava

文献类型： 外文期刊

作者： Tuo, Decai 1 ; Yao, Yuan 1 ; Yan, Pu 1 ; Chen, Xin 1 ; Qu, Feihong 3 ; Xue, Weiqian 3 ; Liu, Jinping 3 ; Kong, Hua 1 ; Guo, Jianchun 1 ; Cui, Hongguang 3 ; Dai, Zhaoji 3 ; Shen, Wentao 1 ;

作者机构： 1.Chinese Acad Trop Agr Sci, Inst Trop Biosci & Biotechnol, Sanya Res Inst, Hainan Inst Trop Agr Resources,Natl Key Lab Trop C, Haikou, Hainan, Peoples R China

2.Chinese Acad Trop Agr Sci, Inst Trop Biosci & Biotechnol, Sanya Res Inst, Hainan Inst Trop Agr Resources,Natl Key Lab Trop C, Sanya, Hainan, Peoples R China

3.Hainan Univ, Sanya Nanfan Res Inst, Sch Trop Agr & Forestry, Haikou, Hainan, Peoples R China

4.Hainan Univ, Sanya Nanfan Res Inst, Sch Trop Agr & Forestry, Sanya, Hainan, Peoples R China

关键词： Cassava; Cassava common mosaic virus; Protein overexpression; Genome editing

期刊名称：PLANT METHODS （ 影响因子：5.1； 五年影响因子：6.1 ）

ISSN：

年卷期： 2023 年 19 卷 1 期

页码：

收录情况： SCI

摘要： BackgroundPlant virus vectors designed for virus-mediated protein overexpression (VOX), virus-induced gene silencing (VIGS), and genome editing (VIGE) provide rapid and cost-effective tools for functional genomics studies, biotechnology applications and genome modification in plants. We previously reported that a cassava common mosaic virus (CsCMV, genus Potexvirus)-based VIGS vector was used for rapid gene function analysis in cassava. However, there are no VOX and VIGE vectors available in cassava.ResultsIn this study, we developed an efficient VOX vector (CsCMV2-NC) for cassava by modifying the CsCMV-based VIGS vector. Specifically, the length of the duplicated putative subgenomic promoter (SGP1) of the CsCMV CP gene was increased to improve heterologous protein expression in cassava plants. The modified CsCMV2-NC-based VOX vector was engineered to express genes encoding green fluorescent protein (GFP), bacterial phytoene synthase (crtB), and Xanthomonas axonopodis pv. manihotis (Xam) type III effector XopAO1 for viral infection tracking, carotenoid biofortification and Xam virulence effector identification in cassava. In addition, we used CsCMV2-NC to deliver single guide RNAs (gMePDS1/2) targeting two loci of the cassava phytoene desaturase gene (MePDS) in Cas9-overexpressing transgenic cassava lines. The CsCMV-gMePDS1/2 efficiently induced deletion mutations of the targeted MePDS with the albino phenotypes in systemically infected cassava leaves.ConclusionsOur results provide a useful tool for rapid and efficient heterologous protein expression and guide RNA delivery in cassava. This expands the potential applications of CsCMV-based vector in gene function studies, biotechnology research, and precision breeding for cassava.

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