当前位置: X-MOL 学术bioRxiv. Plant Biol. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
An acylsugar-deficient Nicotiana benthamiana strain for aphid and whitefly research
bioRxiv - Plant Biology Pub Date : 2021-03-28 , DOI: 10.1101/2020.08.04.237180
Honglin Feng , Lucia Acosta-Gamboa , Lars H. Kruse , Alba Ruth Nava Fereira , Sara Shakir , Hongxing Xu , Garry Sunter , Michael A. Gore , Gaurav D. Moghe , Georg Jander

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 generalist herbivores, including Myzus persicae (green peach aphid), Bemisia tabaci (whitefly), Macrosiphum euphorbiae (potato aphid), Heliothis virescens (tobacco budworm), Trichoplusia ni (cabbage looper), and Helicoverpa zea (corn earworm), 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 acyltransferases, 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. The tested hemipteran and lepidopteran species survived, gained weight, and/or reproduced significantly better on asat2 mutant plants than on wildtype N. benthamiana. Furthermore, both asat1 and asat2 mutations reduced the water content and increased the temperature of leaves, indicating that foliar acylsucroses can protect against desiccation. Two experiments demonstrated the utility of the N. benthamiana asat2 mutant line for insect bioassays. Transmission of turnip mosaic virus by M. persicae was significantly improved by an asat2 mutation. Tobacco rattle virus constructs were used for virus-induced gene silencing of acetylcholinesterase, squalene synthase, toll-like receptor 7, and tubulin-specific chaperon D genes in B. tabaci, an experiment that would have been difficult with wild-type N. benthamiana due to high insect mortality. 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.

中文翻译:

缺乏酰基糖的烟草本氏烟草菌株,用于蚜虫和粉虱研究

烟草(Nicotiana benthamiana)被广泛用作瞬时基因表达的平台和研究植物与病毒相互作用的模型系统。但是,有许多以烟草为食的食草动物,包括桃蚜(Myzus persicae),烟粉虱(Bemisia tabaci)(烟粉虱),大戟(Macrosiphum euphorbiae)(马铃薯蚜虫),Heliothis virescens(烟草芽虫),Trichoplusia ni(卷心菜弯角)和Helicoverpa zea(玉米)。虫)在本塞姆猪笼草上生长较差,限制了其在植物与昆虫相互作用研究中的效用。使用CRISPR / Cas9,我们在两个本氏猪笼草的酰基糖酰基转移酶ASAT1和ASAT2中产生了敲除突变,这有助于昆虫抑制酰基糖的生物合成。尽管asat1突变减少了两个主要的酰基糖的丰度,但是asat2突变却导致了叶酰基酰糖的几乎完全消耗。与野生型本氏烟草相比,在asat2突变植物上测试的半翅目和鳞翅目物种存活,增重和/或繁殖显着更好。此外,asat1和asat2突变均降低了水分含量并提高了叶片的温度,表明叶酰蔗糖可以防止干燥。两项实验证明本氏猪笼草asat2突变株可用于昆虫生物测定。asat2突变显着改善了芜菁花叶病毒对芜菁花叶病毒的传播。烟草摇铃病毒构建体被用于烟粉虱中乙酰胆碱酯酶,角鲨烯合酶,toll​​样受体7和微管蛋白特异性伴侣D基因的病毒诱导基因沉默,这对于野生型本氏烟草难以实现的实验由于昆虫死亡率高。
更新日期:2021-03-29
down
wechat
bug