Screening the function of genes encoding neuropeptides and their receptors in <i>Nilaparvata lugens</i>

doi:10.3785/j.issn.1008-9209.2022.06.292

Journal of Zhejiang University (Agriculture and Life Sciences)

2022, Vol. 48

Issue (6): 766-775 DOI: 10.3785/j.issn.1008-9209.2022.06.292

Research articles

Screening the function of genes encoding neuropeptides and their receptors in Nilaparvata lugens

Siliang WANG^1,²(

),Xumei LUO¹,Chuanxi ZHANG^1,³(

)

^1.Institute of Insect Sciences, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China
^2.College of Agriculture and Biotechnology, Wenzhou Vocational College of Science and Technology, Wenzhou 325006, Zhejiang, China
^3.State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products/Key Laboratory of Biotechnology in Plant Protection of Ministry of Agriculture and Rural Affairs and Zhejiang Province, Institute of Plant Virology, Ningbo University, Ningbo 315211, Zhejiang, China

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Abstract

Neuropeptides are crucial for life activities and environmental fitness of insects, which are potential targets for pest control. The brown planthopper (Nilaparvata lugens) is an important pest on rice in Asia. Thus, functional screening analysis of genes encoding neuropeptides and their receptors in N. lugens were conducted. The cDNA sequences of 41 neuropeptides (containing one alternative splicing transcript) and 44 receptors were verified by polymerase chain reaction (PCR). Four neuropeptide genes (NlCCAP, NlETH, NlOKA and NlPK) and two receptor genes (NlA36 and NlA46) were identified to cause high mortality through RNA interference (RNAi), showing potential in pest control. These genes knockdown resulted in the relative survival rate of adult less than 0.2 when injected with double-stranded RNA (dsRNA) at the third instar, and had the potential to be target genes for pest control. The results of RNAi also demonstrated that neuropeptides, such as crustacean cardioactive peptide (CCAP), ecdysis triggering hormone (ETH) and bursicon were conserved in physiological function, while other neuropeptides (eg. prothoracicotropic hormone, eclosion hormone) which related to ecdysis in other insects were undefined in physiological function in N. lugens. Our study provides a basis for further exploration of neuropeptides’ physiological function in N. lugens.

Key words： Nilaparvata lugens neuropeptide RNA interference ecdysis

Received: 29 June 2022 Published: 27 December 2022

CLC:

S 433.3

Corresponding Authors: Chuanxi ZHANG E-mail: wsl512935@163.com;chxzhang@zju.edu.cn

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Cite this article:

Siliang WANG,Xumei LUO,Chuanxi ZHANG. Screening the function of genes encoding neuropeptides and their receptors in Nilaparvata lugens. Journal of Zhejiang University (Agriculture and Life Sciences), 2022, 48(6): 766-775.

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https://www.zjujournals.com/agr/10.3785/j.issn.1008-9209.2022.06.292 OR https://www.zjujournals.com/agr/Y2022/V48/I6/766

褐飞虱神经肽及其受体基因的功能筛查

神经肽对昆虫的生命活动和环境适应性具有重要影响，是害虫防治的潜在靶标。褐飞虱（Nilaparvata lugens）是亚洲地区一种重要的水稻害虫。本研究对褐飞虱神经肽及其受体基因的功能进行了筛查，通过聚合酶链反应扩增并验证得到了41个神经肽基因（含1个可变剪接本）和44个受体基因。通过RNA干扰方法，得到沉默后造成高死亡率的4个神经肽基因（NlCCAP、NlETH、NlOKA和NlPK）和2个受体基因（NlA36和NlA46），这些基因在褐飞虱3龄若虫阶段注射双链RNA（double-stranded RNA, dsRNA）进行沉默后，成虫相对存活率＜0.2，具有成为害虫防治靶标的潜力。RNA干扰结果表明：几种蜕皮前后行为调控神经肽，如甲壳类动心肽、蜕皮诱导激素、鞣化激素的基因与其他昆虫研究中报道的功能相似，而另一些与蜕皮调控相关的神经肽（如促前胸腺激素、羽化激素）在褐飞虱中的生理功能则不明显。本研究为褐飞虱神经肽生理功能的深入探索奠定了基础。

关键词： 褐飞虱, 神经肽, RNA干扰, 蜕皮

Table 1 RNAi effects of neuropeptide genes in N. lugens

Table 2 RNAi effects of neuropeptide receptor genes in N. lugens

Fig. 1 Survival rates and phenotypes of N. lugens with NlETH knockdown after RNAi

Fig. 2 Survival rates and phenotypes of N. lugens with NlCCAP knockdown after RNAiA. Survival rates of N. lugens when injected with dsNlCCAP at different instars; B. Phenotypes of N. lugens with NlCCAP knockdown after RNAi.

Fig. 3 Expression level changes of genes encoding two subunits of bursicon after injected with different dsRNAs

Fig. 4 Survival rates and phenotypes of N. lugens with Nlburs-a and Nlburs-b knockdown after RNAiA. Survival rates of N. lugens when injected with dsNlburs-a and dsNlburs-b at different instars; B. Phenotypes of N. lugens with Nlburs-a and Nlburs-b knockdown after RNAi.

Fig. 5 Effects of bursicon knockdown on phenotypes (A) and wing-unstretched rates of adults (B) in different instars of N. lugens

Fig. 6 Survival rates and phenotypes of N. lugens with NlA46 knockdown after RNAi


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