Ahmad Shakil, Jamil Momana, Jaworski Coline C, Luo Yanping
School of Plant Protection, Hainan University, Haikou, Hainan, China.
Université Côte d'Azur, INRAE, CNRS, UMR ISA, Nice, France.
Front Physiol. 2023 Jan 17;14:1112548. doi: 10.3389/fphys.2023.1112548. eCollection 2023.
Irradiation can be used as an insect pest management technique to reduce post-harvest yield losses. It causes major physiological changes, impairing insect development and leading to mortality. This technique is used to control the melon fly , a major pest of Cucurbitaceae in Asia. Here, we applied irradiation to melon fly eggs, and the larvae emerged from irradiated eggs were used to conduct comparative transcriptomics and thereby identify key genes involved in the development and survival. We found 561 upregulated and 532 downregulated genes in irradiated flies compared to non-irradiated flies. We also observed abnormal small-body phenotypes in irradiated flies. By screening the 532 downregulated genes, we selected eight candidate genes putatively involved in development based in described functions in public databases and in the literature. We first established the expression profile of each candidate gene. Using RNA interference (RNAi), we individually knocked down each gene in third instar larvae and measured the effects on development. The knockdown of ecdysone-inducible gene controlling life stage transitions-led to major body size reductions in both pupae and adults. The knockdown of the tyrosine-protein kinase-like tok (Tpk-tok) caused severe body damage to larvae, characterized by swollen and black body parts. Adults subject to knockdown of the eclosion hormone () failed to shed their old cuticle which remained attached to their bodies. However, no obvious developmental defects were observed following the knockdown of the heat shock protein 67B1-like (Hsp67), the insulin receptor (), the serine/threonine-protein kinase Nek4 (Nek4), the tyrosine-protein kinase transmembrane receptor Ror () and the probable insulin-like peptide 1 (). We argue that irradiation can be successfully used not only as a pest management technique but also for the screening of essential developmental genes in insects comparative transcriptomics. Our results demonstrate that and are essential for the development of melon fly and could therefore be promising candidates for the development of RNAi-based pest control strategies.
辐照可作为一种害虫管理技术,用于减少收获后产量损失。它会引起重大生理变化,损害昆虫发育并导致死亡。该技术用于防治瓜实蝇,这是亚洲葫芦科作物的主要害虫。在此,我们对瓜实蝇卵进行辐照,将辐照卵孵化出的幼虫用于进行比较转录组学研究,从而鉴定参与发育和存活的关键基因。与未辐照的果蝇相比,我们在辐照果蝇中发现了561个上调基因和532个下调基因。我们还在辐照果蝇中观察到了异常的小体型表型。通过筛选这532个下调基因,我们根据公共数据库和文献中描述的功能,选择了八个可能参与发育的候选基因。我们首先建立了每个候选基因的表达谱。使用RNA干扰(RNAi),我们在三龄幼虫中分别敲低每个基因,并测量对发育的影响。敲低控制生命阶段转变的蜕皮激素诱导基因,导致蛹和成虫的体型大幅减小。敲低酪氨酸蛋白激酶样tok(Tpk - tok)会对幼虫造成严重的身体损伤,其特征是身体部位肿胀和变黑。敲低羽化激素()的成虫无法蜕去仍附着在其身体上的旧表皮。然而,在敲低热休克蛋白67B1样(Hsp67)、胰岛素受体()、丝氨酸/苏氨酸蛋白激酶Nek4(Nek4)、酪氨酸蛋白激酶跨膜受体Ror()和可能的胰岛素样肽1()后,未观察到明显的发育缺陷。我们认为辐照不仅可以成功用作害虫管理技术,还可用于通过比较转录组学筛选昆虫发育必需基因。我们的结果表明,和对瓜实蝇的发育至关重要,因此可能是基于RNAi的害虫控制策略开发的有前途的候选基因。