Optimization of dietary RNA interference delivery to western flower thrips Frankliniella occidentalis and onion thrips Thrips tabaci.
Optimization of dietary RNA interference delivery to western flower thrips Frankliniella occidentalis and onion thrips Thrips tabaci.
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DOI：
10.1002/arch.21645
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发表时间：
2020-03
影响因子：
2.2
通讯作者：
Whitten, Miranda M. A.
中科院分区：
文献类型：
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作者：
Andongma, Awawing A.;Greig, Carolyn;Dyson, Paul J.;Flynn, Natasha;Whitten, Miranda M. A.
在昆虫逆向遗传学中，通过饮食传递干扰rna是一种实用的方法，适用于非模式物种，如蓟马，其体型小，摄食行为限制了其他传递方法的使用。在实验室环境中，不合适的饲料可能会混淆RNA干扰（RNAi）表型的解释，然而精心配制的人工饲料可以最大限度地减少实验变异性，减少对昆虫处理的需求，并可以进一步用于诸如传递表达双链RNA （dsRNA）的重组细菌等角色。本研究为两种重要的害虫蓟马——西方花蓟马（Frankliniella occidentalis）和烟蓟马（thrips tabaci）开发了用于口服dsRNA的人工饲料，目的是(a)刺激摄食行为，(b)支持表达dsRNA的共生细菌的最佳生长速度，(c)在营养上支持蓟马足够的时间来观察RNAi表型。评估了人工饲料摄入“裸”dsRNA或表达dsRNA的共生体以及通过寄主植物摄取dsRNA的效果。与先前发表的饲料配方相比，基于色氨酸、酵母和大豆的新组合在提高饲养和寿命方面更胜一筹。然而，简单地将“裸”dsRNA添加到人工饮食中是一种不可靠的RNAi递送形式，因为dsRNA会降解。通过寄主植物传递更为成功，并且新的饲料配方适合共生体介导的dsRNA传递，我们认为这是大规模敲低实验中最方便的方法。因此，本研究为蓟马饲养、膳食RNAi递送提供了替代方法，并深入了解了在非模式昆虫中执行膳食RNAi的挑战。两种蓟马双链RNA （dsRNA）口服递送方法的比较。我们将重点研究提高饲养活性和寿命、支持细菌共生介导的RNA干扰的日粮配方，并与基于植物的dsRNA传递和直接摄入方法进行比较。为优化两种蓟马的饲养和生存而开发的人工饲料是细菌共生体介导的dsRNA传递的理想选择。RNAi也通过宿主植物对dsRNA的摄取来实现，但蓟马的摄食活动可以降解裸合成的dsRNA。
In insect reverse genetics, dietary delivery of interfering RNAs is a practical approach in nonmodel species, such as thrips, whose small size, and feeding behavior restricts the use of other delivery methods. In a laboratory context, an unsuitable diet could confound the interpretation of an RNA interference (RNAi) phenotype, however well‐formulated artificial diets can minimize experimental variability, reduce the need for insect handling, and can further be used for roles, such as delivering double‐strand RNA (dsRNA)‐expressing recombinant bacteria. In this study, artificial diets for oral delivery of dsRNA were developed for two important pest thrips species, western flower thrips (Frankliniella occidentalis) and onion thrips (Thrips tabaci), with the goal of (a) stimulating feeding behavior, (b) supporting optimal growth rates of dsRNA‐expressing symbiotic bacteria, and (c) nutritionally supporting the thrips for sufficient periods to observe RNAi phenotypes. The efficacy of artificial diets for ingesting “naked” dsRNA or dsRNA‐expressing symbionts and dsRNA delivery via host plant uptake was evaluated. Compared with previously published diet formulations, new combinations based on tryptone, yeast, and soy were superior for enhancing feeding and longevity. However, simply adding “naked” dsRNA to an artificial diet was an unreliable form of RNAi delivery in our hands due to dsRNA degradation. Delivery via host plants was more successful, and the new diet formulation was suitable for symbiont‐mediated dsRNA delivery, which we believe is the most convenient approach for large‐scale knockdown experiments. This study, therefore, provides alternative methodologies for thrips rearing, dietary RNAi delivery, and insights into the challenges of performing dietary RNAi in nonmodel insects. A comparison of double‐strand RNA (dsRNA) oral delivery methods for two thrips species. We focus on diet formulations that enhance feeding activity and longevity and support bacterial symbiont‐mediated RNA interference, and compare with plant‐based dsRNA delivery and direct ingestion methods. Artificial diets developed to optimize feeding and survival in two thrips species are ideal for bacterial symbiont‐mediated dsRNA delivery. RNAi is also achieved via host plant dsRNA uptake, but thrips feeding activity can degrade naked synthetic dsRNA.
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影响因子：
3.7
作者：
Ghosh SK;Hunter WB;Park AL;Gundersen-Rindal DE
通讯作者：
Gundersen-Rindal DE
影响因子：
4.6
作者：
Niu X;Kassa A;Hu X;Robeson J;McMahon M;Richtman NM;Steimel JP;Kernodle BM;Crane VC;Sandahl G;Ritland JL;Presnail JK;Lu AL;Wu G
通讯作者：
Wu G
影响因子：
3.3
作者：
Facey PD;Méric G;Hitchings MD;Pachebat JA;Hegarty MJ;Chen X;Morgan LV;Hoeppner JE;Whitten MM;Kirk WD;Dyson PJ;Sheppard SK;Del Sol R
通讯作者：
Del Sol R
影响因子：
2.2
作者：
Allen, Margaret L.;Walker, William B., III
通讯作者：
Walker, William B., III
DOI：
10.1006/smvy.1994.1012
发表时间：
1994-04-01
期刊：
SEMINARS IN VIROLOGY
影响因子：
--
作者：
GOLDBACH, R;PETERS, D
通讯作者：
PETERS, D