Simultaneous CRISPR/Cas9-mediated editing of cassava eIF4E isoforms nCBP-1 and nCBP-2 reduces cassava brown streak disease symptom severity and incidence.
Simultaneous CRISPR/Cas9-mediated editing of cassava eIF4E isoforms nCBP-1 and nCBP-2 reduces cassava brown streak disease symptom severity and incidence.
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DOI：
10.1111/pbi.12987
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发表时间：
2019-03
影响因子：
13.8
通讯作者：
Bart RS
中科院分区：
文献类型：
--
作者：
Gomez MA;Lin ZD;Moll T;Chauhan RD;Hayden L;Renninger K;Beyene G;Taylor NJ;Carrington JC;Staskawicz BJ;Bart RS
木薯褐条病（CBSD）是东非和中非木薯产量的主要制约因素，并威胁西非的生产。CBSD是由属于马铃薯Y病毒科、伊普莫病毒属的两种正义RNA病毒引起的：木薯褐条病毒（CBSV）和乌干达木薯褐条病毒（UCBSV）。由马铃薯Y病毒科引起的疾病需要病毒基因组连接蛋白（VPg）和宿主真核翻译起始因子4 E（eIF 4 E）亚型的相互作用。木薯编码五种eIF 4 E蛋白：eIF 4 E、eIF（iso）4 E-1、eIF（iso）4 E-2、novel cap-binding protein-1（nCBP-1）和nCBP-2。蛋白质-蛋白质相互作用实验一致地发现VPg蛋白与木薯nCBPs相关。采用CRISPR/Cas9介导的基因组编辑在木薯栽培品种60444中产生ncbp-1、ncbp-2和ncbp-1/ncbp-2突变体。用CBSV攻击表明，ncbp-1/ncbp-2突变体表现出延迟和减弱的CBSD气生症状，以及降低的严重程度和贮藏根坏死的发生率。相对于野生型对照，抑制的疾病症状与储存根中降低的病毒滴度相关。我们的研究结果证明了在木薯中同时修饰多个基因以实现对CBSD的耐受性的能力。未来的研究将调查剩余的eIF 4 E亚型对CBSD的贡献，并将这些知识转化为保护木薯免受疾病的优化策略。
Cassava brown streak disease (CBSD) is a major constraint on cassava yields in East and Central Africa and threatens production in West Africa. CBSD is caused by two species of positive‐sense RNA viruses belonging to the family Potyviridae, genus Ipomovirus: Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV). Diseases caused by the family Potyviridae require the interaction of viral genome‐linked protein (VPg) and host eukaryotic translation initiation factor 4E (eIF4E) isoforms. Cassava encodes five eIF4E proteins: eIF4E, eIF(iso)4E‐1, eIF(iso)4E‐2, novel cap‐binding protein‐1 (nCBP‐1), and nCBP‐2. Protein–protein interaction experiments consistently found that VPg proteins associate with cassava nCBPs. CRISPR/Cas9‐mediated genome editing was employed to generate ncbp‐1, ncbp‐2, and ncbp‐1/ncbp‐2 mutants in cassava cultivar 60444. Challenge with CBSV showed that ncbp‐1/ncbp‐2 mutants displayed delayed and attenuated CBSD aerial symptoms, as well as reduced severity and incidence of storage root necrosis. Suppressed disease symptoms were correlated with reduced virus titre in storage roots relative to wild‐type controls. Our results demonstrate the ability to modify multiple genes simultaneously in cassava to achieve tolerance to CBSD. Future studies will investigate the contribution of remaining eIF4E isoforms on CBSD and translate this knowledge into an optimized strategy for protecting cassava from disease.
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影响因子：
14.9
作者：
Goujon M;McWilliam H;Li W;Valentin F;Squizzato S;Paern J;Lopez R
通讯作者：
Lopez R
影响因子：
46.9
作者：
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影响因子：
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作者：
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影响因子：
7.2
作者：
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通讯作者：
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影响因子：
3.7
作者：
Ashby JA;Stevenson CE;Jarvis GE;Lawson DM;Maule AJ
通讯作者：
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