褐飞虱NlTgo基因的克隆及功能研究
收稿日期: 2016-03-01
修回日期: 2016-04-21
网络出版日期: 2016-11-10
基金资助
现代农业产业技术体系专项(CARS-1-18);国家自然科学基金资助项目(31201512)
Molecular Cloning and Functional Analysis of NlTgo in the Rice Brown Planthopper, Nilaparvata lugens (Hemiptera: Delphacidae)
Received date: 2016-03-01
Revised date: 2016-04-21
Online published: 2016-11-10
转录因子Tango(Tgo)在昆虫神经元发生、血细胞生成、性别决定、肠道发育等过程中发挥重要作用。克隆了褐飞虱Tango基因(NlTgo),应用荧光定量PCR和RNAi探索了NlTgo在褐飞虱中的表达动态和生物学功能。结果表明,NlTgo的开放阅读框为2007 bp,推测编码669个氨基酸残基。多序列比对表明NlTgo与已知的Tgo高度同源,其中与人体虱(Pediculus humanus corporis) Tgo的一致性达68%。系统进化分析表明,NlTgo与豌豆长管蚜(Acyrthosiphon pisum)、始红蝽(Pyrrhocoris apterus)和臭虫(Cimex lectularius)的Tgo在系统发育树中形成一个类群。时空表达谱表明NlTgo在褐飞虱1龄、2龄时期高表达,在卵期表达量较低;在卵巢中表达量最高,在体壁的表达量较低。RNAi结果表明,注射dsNlTgo 后4 d,NlTgo 的表达量较空白对照组显著降低了77%,并导致若虫不能正常蜕皮而死亡。其中,处理5 d后褐飞虱的存活率仅为23%,显著低于注射dsGFP的对照组(98%)。结果显示,NlTgo与褐飞虱的生长发育有关,可作为防治褐飞虱的潜在靶标。
陈龙飞, 万品俊, 王渭霞, 傅强, 朱廷恒 . 褐飞虱NlTgo基因的克隆及功能研究[J]. 中国水稻科学, 2016 , 30(6) : 653 -660 . DOI: 10.16819/j.1001-7216.2016.6032
Transcriptional regulator Tango (Tgo) plays crucial roles in insect development, including neurogenesis, hematopoiesis, sex determination and gut development. In this study, a full-length cDNA of Tango was cloned in Nilaparvata lugens. qRT-PCR and RNA interference (RNAi) were further used to analyze the expression pattern and function role, respectively. Our results showed that NlTgo contained a 2007-bp open reading frame (ORF), encoding 669 amino acid residues. Sequence alignment showed that NlTgo shared an identity of 68% with Pediculus humanus. Phylogenetic analysis suggested that NlTgo was closely related to the Tango proteins from Acyrthosiphon pisum, Pyrrhocoris apterus and Cimex lectularius. Expression profile revealed that NlTgo expression was higher in the first- and second-instar larvae than that in eggs. Furthermore, the expression level of NlTgo was higher in ovary than that in integument. Knocking down of NlTgo, in the 4th-instar nymph, was performed by double-stranded RNA (dsRNA) targeting NlTgo. It was found that the expression level of NlTgo, 4 days after injection, was significantly decreased by 77%, compared with control. Furthermore, nymphs died due to the abnormal molting, and the survival rate was only 23%, significantly lower than control group (98%). The results suggest that NlTgo is involved in the development of N.lugens and can serve as a potential target for controlling the brown planthopper.
Key words： Nilaparvata lugens; Tango; RNAi; molting; pest control
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