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基于柑橘衰退病毒的RNA干扰技术在柑橘植株中可诱导柑桔木虱(柑橘黄龙病韧皮部汁液吸食性昆虫传播介体)的基因沉默。

Citrus tristeza virus-based RNAi in citrus plants induces gene silencing in Diaphorina citri, a phloem-sap sucking insect vector of citrus greening disease (Huanglongbing).

作者信息

Hajeri Subhas, Killiny Nabil, El-Mohtar Choaa, Dawson William O, Gowda Siddarame

机构信息

Department of Plant Pathology, University of Florida, Citrus Research and Education Center, 700 Experiment Station Road, Lake Alfred, FL 33850, USA.

Department of Entomology and Nematology, University of Florida, Citrus Research and Education Center, 700 Experiment Station Road, Lake Alfred, FL 33850, USA.

出版信息

J Biotechnol. 2014 Apr 20;176:42-9. doi: 10.1016/j.jbiotec.2014.02.010. Epub 2014 Feb 23.

Abstract

A transient expression vector based on Citrus tristeza virus (CTV) is unusually stable. Because of its stability it is being considered for use in the field to control Huanglongbing (HLB), which is caused by Candidatus Liberibacter asiaticus (CLas) and vectored by Asian citrus psyllid, Diaphorina citri. In the absence of effective control strategies for CLas, emphasis has been on control of D. citri. Coincident cohabitation in phloem tissue by CLas, D. citri and CTV was exploited to develop a novel method to mitigate HLB through RNA interference (RNAi). Since CTV has three RNA silencing suppressors, it was not known if CTV-based vector could induce RNAi in citrus. Yet, expression of sequences targeting citrus phytoene desaturase gene by CTV-RNAi resulted in photo-bleaching phenotype. CTV-RNAi vector, engineered with truncated abnormal wing disc (Awd) gene of D. citri, induced altered Awd expression when silencing triggers ingested by feeding D. citri nymphs. Decreased Awd in nymphs resulted in malformed-wing phenotype in adults and increased adult mortality. This impaired ability of D. citri to fly would potentially limit the successful vectoring of CLas bacteria between citrus trees in the grove. CTV-RNAi vector would be relevant for fast-track screening of candidate sequences for RNAi-mediated pest control.

摘要

一种基于柑橘衰退病毒(CTV)的瞬时表达载体异常稳定。由于其稳定性,它正被考虑用于田间防治黄龙病(HLB),该病由亚洲韧皮杆菌(CLas)引起,并由柑橘木虱传播。在缺乏针对CLas的有效控制策略的情况下,重点一直放在控制柑橘木虱上。利用CLas、柑橘木虱和CTV在韧皮部组织中的共生关系,开发了一种通过RNA干扰(RNAi)减轻黄龙病的新方法。由于CTV有三种RNA沉默抑制因子,尚不清楚基于CTV的载体是否能在柑橘中诱导RNAi。然而,CTV-RNAi靶向柑橘八氢番茄红素去饱和酶基因的序列表达导致了光漂白表型。用柑橘木虱截短的异常翅盘(Awd)基因构建的CTV-RNAi载体,在柑橘木虱若虫摄食沉默触发物时,诱导Awd表达改变。若虫中Awd减少导致成虫出现翅畸形表型,并增加成虫死亡率。柑橘木虱飞行能力的这种受损可能会限制CLas细菌在果园柑橘树之间的成功传播。CTV-RNAi载体将有助于快速筛选用于RNAi介导的害虫防治的候选序列。

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