褐飞虱26S蛋白酶体非ATPase调节亚基4基因NlPSMD4作为防治靶点的潜力研究*
Potential for targeting the 26S proteasome non-ATPase regulatory subunit four gene NlPSMD4 for the control of Nilaparvata lugens
施书棋** 杨江涛 沈博昱 郝培应*** 俞晓平***
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DOI：10.7679/j.issn.20951353.2026.110
作者单位：中国计量大学生命科学学院，浙江省生物计量及检验检疫技术重点实验室，杭州 310018
中文关键词：褐飞虱；PSMD4；26S蛋白酶体；RNA干扰；害虫防治
英文关键词：Nilaparvata lugens; PSMD4; 26S proteasome; RNA interference; pest control
中文摘要：【目的】 褐飞虱Nilaparvata lugens是对水稻极具破坏性的农业害虫。泛素-蛋白酶体系统（Ubiquitin proteasome system，UPS）参与昆虫多种生理过程。本研究旨在探究UPS的核心组分26S蛋白酶体非ATP酶调节亚基4（NlPSMD4）在褐飞虱生长发育与生殖中的功能，并评估其作为RNA干扰（RNA interference，RNAi）防控靶点的潜力。【方法】 利用qRT-PCR技术解析NlPSMD4的时空表达特征；通过显微注射dsRNA对若虫和成虫分别进行NlPSMD4基因的RNA干扰，系统测定其致死效应、产卵量及卵孵化率；检测NlPSMD4基因RNA干扰后卵黄原蛋白（Vg）、保幼激素酸甲基转移酶（JHAMT）以及蜕皮激素受体（EcR）相关基因的表达量变化，以揭示其作用机制。【结果】 NlPSMD4在褐飞虱各发育阶段及组织中均有表达，其中卵期第2天及卵巢组织丰度最高。RNAi结果表明，干扰NlPSMD4会导致褐飞虱严重死亡：dsNlPSMD4注射处理后第11天若虫存活率仅为3.33%，雄虫与雌虫存活率分别骤降至4.00%（10 d）和4.17%（14 d）。生殖力测定显示，雌雄双性别干扰组（dsNlPSMD4♀×dsNlPSMD4♂）总产卵量降至62粒，孵化率降至25.65%；雄虫单性别干扰亦显著抑制孵化率（41.93%，P＜0.05）。产卵量与孵化率相关的基因表达水平检测发现，NlPSMD4基因的RNA干扰破坏了激素与生殖信号稳态： JHAMT基因在雌成虫中的表达显著上调（P＜0.000 1）；EcR基因在雄虫中显著上调（P＜0.000 1）；雌虫Vg基因表达量激增约15倍（20.07），但产卵量和孵化率没有增加，反而降低。【结论】 本研究证实NlPSMD4在褐飞虱生存及生殖方面发挥重要作用，其表达量下降会引发严重的致死与致不育效应。因此，NlPSMD4有望成为基于RNA干扰的褐飞虱防控新的靶标。
英文摘要：[Aim] To investigate the function of the 26S proteasome non-ATPase regulatory subunit 4 (NlPSMD4), a core component of the UPS, in the growth, development and reproduction of Nilaparvata lugens, and evaluate its potential as a target for RNA interference (RNAi)-based control. [Methods] The spatiotemporal expression profiles of the NlPSMD4 gene were characterized using qRT-PCR. Gene silencing was performed by microinjecting dsRNA into nymphs and adults. Following RNAi treatment, mortality, fecundity, and egg hatching rates were systematically measured. Furthermore, the mRNA expression levels of vitellogenin (Vg), juvenile hormone acid methyltransferase (JHAMT) and ecdysone receptor (EcR) genes, were analyzed to elucidate the underlying molecular mechanisms. [Results] NlPSMD4 was expressed in all developmental stages and tissues, with the highest abundance observed in 2-day-old eggs and ovarian tissues. RNAi experiments demonstrated that silencing NlPSMD4 caused severe mortality; nymph survival dropped to 3.33% by day 11 post-treatment, and the survival rate of males and females declined to 4.00% and 4.17%, respectively, on days 10 and 14. RNAi treatment of both sexes (dsNlPSMD4♀ × dsNlPSMD4♂) caused the number of eggs to decrease to 62 per female, and the hatching rate to decrease to 25.65%. RNAi treatment of males only (dsGFP♀ × dsNlPSMD4♂) also significantly reduced the hatching rate of eggs to 41.93%. Gene expression analysis indicates that RNAi treatment disrupts hormonal and reproductive signaling homeostasis. mRNA expression of the JHAMT gene was significantly up-regulated in female adults, whereas that of the EcR gene was significantly up-regulated in males. Although the mRNA expression of the Vg gene in females surged 15-fold (20.07), the fecundity and hatching rates of eggs did not increase, but rather declined. [Conclusion] This study confirms that the NlPSMD4 gene plays a critical role in the survival and reproduction of the BPH. Loss of NlPSMD4 function results in significant lethality and sterility. Therefore, the NlPSMD4 gene is a promising target for the RNAi-based management of BPH.