Division of Nematology, ICAR-Indian Agricultural Research Institute, New Delhi, India.
Functional Genomics Center Zurich, University of Zurich/ETH Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland.
Int J Biol Macromol. 2024 Jan;254(Pt 1):127666. doi: 10.1016/j.ijbiomac.2023.127666. Epub 2023 Oct 25.
The spotted pod borer, Maruca vitrata (Lepidoptera: Crambidae) is a destructive insect pest that inflicts significant productivity losses on important leguminous crops. Unravelling insect proteomes is vital to comprehend their fundamental molecular mechanisms. This research delved into the proteome profiles of four distinct stages -three larval and pupa of M. vitrata, utilizing LC-MS/MS label-free quantification-based methods. Employing comprehensive proteome analysis with fractionated datasets, we mapped 75 % of 3459 Drosophila protein orthologues out of which 2695 were identified across all developmental stages while, 137 and 94 were exclusive to larval and pupal stages respectively. Cluster analysis of 2248 protein orthologues derived from MaxQuant quantitative dataset depicted six clusters based on expression pattern similarity across stages. Consequently, gene ontology and protein-protein interaction network analyses using STRING database identified cluster 1 (58 proteins) and cluster 6 (25 proteins) associated with insect immune system and lipid metabolism. Furthermore, qRT-PCR-based expression analyses of ten selected proteins-coding genes authenticated the proteome data. Subsequently, functional validation of these chosen genes through gene silencing reduced their transcript abundance accompanied by a marked increase in mortality among dsRNA-injected larvae. Overall, this is a pioneering study to effectively develop a proteome atlas of M. vitrata as a potential resource for crop protection programs.
点斑豆芫菁,Maruca vitrata(鳞翅目:草螟科)是一种具有破坏性的昆虫害虫,会对重要豆科作物造成重大生产力损失。揭示昆虫蛋白质组对于理解其基本分子机制至关重要。本研究利用 LC-MS/MS 无标记定量方法,深入研究了四个不同阶段——三个幼虫和蛹期的 M. vitrata 的蛋白质组图谱。通过对分馏数据集进行全面的蛋白质组分析,我们绘制了 3459 个果蝇蛋白直系同源物中的 75%,其中 2695 个在所有发育阶段都被识别出来,而 137 个和 94 个分别是幼虫和蛹期所特有的。基于 MaxQuant 定量数据集的 2248 个蛋白直系同源物的聚类分析根据各阶段表达模式的相似性,描绘了六个聚类。因此,使用 STRING 数据库进行的基因本体和蛋白质-蛋白质相互作用网络分析,确定了与昆虫免疫系统和脂质代谢相关的簇 1(58 个蛋白)和簇 6(25 个蛋白)。此外,通过 qRT-PCR 对十个选定蛋白编码基因的表达分析验证了蛋白质组数据。随后,通过基因沉默对这些选定基因进行功能验证,降低了它们的转录丰度,同时 dsRNA 注射幼虫的死亡率显著增加。总的来说,这是一项开创性的研究,成功地为 M. vitrata 开发了蛋白质组图谱,作为作物保护计划的潜在资源。