An improved Nicotiana benthamiana strain for aphid and whitefly research
An improved Nicotiana benthamiana strain for aphid and whitefly research
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DOI：
10.1101/2020.08.04.237180
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发表时间：
2020-08
期刊：
影响因子：
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通讯作者：
Honglin Feng;Lucia M. Acosta-Gamboa;Lars H. Kruse;Alba Ruth Nava Fereira;Sara Shakir;Hong-xing Xu;G. Sunter;M. Gore;G. Moghe;G. Jander
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作者：
Honglin Feng;Lucia M. Acosta-Gamboa;Lars H. Kruse;Alba Ruth Nava Fereira;Sara Shakir;Hong-xing Xu;G. Sunter;M. Gore;G. Moghe;G. Jander
本氏烟草（Nicotiana benthamiana）被广泛用作瞬时基因表达的平台和研究植物-病毒相互作用的模型系统。然而，许多食烟昆虫，如桃蚜（绿色桃蚜）和烟粉虱（粉虱），在N. benthamiana，限制了其在植物-昆虫相互作用研究中的效用。使用CRISPR/Cas9，我们在两个N.本塞姆氏菌酰基糖酰基转移酶基因ASAT 1和ASAT 2，其有助于昆虫忌避性酰基蔗糖的生物合成。而ASAT 1突变减少了两个主要的酰基蔗糖的丰度，ASAT 2突变几乎完全耗尽叶酰基蔗糖。两个M. persicae和B.烟粉虱在asat 2突变体上的存活和繁殖显著优于野生型烟粉虱。本萨米亚纳此外，ASAT 1和ASAT 2突变体降低了叶片的含水量，提高了叶片的温度，表明叶片酰基蔗糖可以防止脱水。ASAT 2突变体对蚜虫和粉虱性能的改善将使使用有效的瞬时过表达和基因表达沉默系统成为可能，这些系统可用于N. benthamiana研究植物与昆虫的相互作用。此外，ASAT 2突变株系中酰基糖的缺失将简化用于来自其他茄科的酰基糖生物合成基因的功能分析的瞬时表达测定。
Nicotiana benthamiana is used extensively as a platform for transient gene expression and as a model system for studying plant-virus interactions. However, many tobacco-feeding insects, e.g. Myzus persicae (green peach aphids) and Bemisia tabaci (whiteflies), grow poorly on N. benthamiana, limiting its utility for research on plant-insect interactions. Using CRISPR/Cas9, we generated knockout mutations in two N. benthamiana acylsugar acyltransferase genes, ASAT1 and ASAT2, which contribute to the biosynthesis of insect-deterrent acylsucroses. Whereas ASAT1 mutations reduced the abundance of two predominant acylsucroses, ASAT2 mutations caused almost complete depletion of foliar acylsucroses. Both M. persicae and B. tabaci survived and reproduced significantly better on asat2 mutant plants than on wildtype N. benthamiana. Furthermore, ASAT1 and ASAT2 mutations reduced the water content and increased the temperature of leaves, indicating that foliar acylsucroses can protect against desiccation. Improved aphid and whitefly performance on ASAT2 mutants will make it possible to use the efficient transient overexpression and gene expression silencing systems that are available for N. benthamiana to study plant-insect interactions. Additionally, the absence of acylsugars in ASAT2 mutant lines will simplify transient expression assays for the functional analysis of acylsugar biosynthesis genes from other Solanaceae.