Harith-Fadzilah Nazmi, Nihad Mohammad, AlSaleh Mohammed Ali, Bazeyad Abdulqader Yaslam, Pandurangan Subash-Babu, Munawar Kashif, Vidyawan Arya, Alharbi Hattan A, Jakše Jernej, Pain Arnab, Antony Binu
School of Agriculture Sciences and Biotechnology, Faculty of Bioresources and Food Industry, Universiti Sultan Zainal Abidin, Besut 22200, Malaysia.
Department of Plant Protection, Center for Chemical Ecology and Functional Genomics, College of Food and Agricultural Sciences, King Saud University, Riyadh 11451, Saudi Arabia.
Insects. 2025 Apr 17;16(4):421. doi: 10.3390/insects16040421.
The red palm weevil, , is a destructive, invasive pest to a diverse range of palm plantations globally. Commonly used broad-range chemical insecticides for insect control pose high risks to non-target organisms, humans, and the environment. A bio-rational approach of screening natural small-molecule inhibitors that specifically target proteins critical to its life processes can pave the way for developing novel bioinsecticides. Digestive enzymes (DEs), which impair feeding on plants (herbivory), are promising targets. We generated de novo transcriptomes, annotated DE-related genes from the gut and abdomen, manually annotated the DE gene family from the recently available genome and our transcriptome data, and reported 34 glycosidases, 85 lipases, and 201 proteases. We identified several tandem duplicates and allelic variants from the lipase and protease families, notably, 10 RferLip and 21 RferPro alleles, which emerged primarily through indels and single-site substitution. These alleles may confer enhanced digestive lipolysis and proteolysis. Phylogenetic analyses identified and classified different subfamilies of DEs and revealed close evolutionary relationships with other coleopterans. We assessed select candidate DEs' activity and the potential for inhibition in silico to better understand the herbivory arsenal. In silico analysis revealed that the selected enzymes exhibited similar ligand-binding affinity to their corresponding substrate, except for protease aminopeptidase N, RferPro40, which exhibited poorer affinity to the inhibitor bestatin. Overall, our study serves as a foundation for further functional analysis and offers a novel target for the development of a novel bio-rational insecticide for .
红棕象甲是全球多种棕榈种植园的一种具有破坏性的入侵害虫。常用的广谱化学杀虫剂对昆虫进行防治时，会对非目标生物、人类和环境构成高风险。一种筛选特异性靶向其生命过程关键蛋白质的天然小分子抑制剂的生物合理方法，可为开发新型生物杀虫剂铺平道路。消化酶（DEs）会损害对植物的取食（食草行为），是很有前景的靶点。我们从头生成了转录组，注释了来自肠道和腹部的与DE相关的基因，根据最近可得的基因组和我们的转录组数据手动注释了DE基因家族，并报告了34种糖苷酶、85种脂肪酶和201种蛋白酶。我们从脂肪酶和蛋白酶家族中鉴定出了几个串联重复和等位基因变体，特别是10个红棕象甲脂肪酶（RferLip）和21个红棕象甲蛋白酶（RferPro）等位基因，它们主要通过插入缺失和单碱基替换出现。这些等位基因可能赋予增强的消化性脂肪分解和蛋白水解能力。系统发育分析鉴定并分类了DEs的不同亚家族，并揭示了与其他鞘翅目昆虫的密切进化关系。我们评估了选定候选DEs的活性以及在计算机模拟中的抑制潜力，以更好地了解食草武器库。计算机模拟分析表明，除了蛋白酶氨肽酶N（RferPro40）对抑制剂贝司他汀的亲和力较差外，所选酶对其相应底物表现出相似的配体结合亲和力。总体而言，我们的研究为进一步的功能分析奠定了基础，并为开发针对红棕象甲的新型生物合理杀虫剂提供了一个新靶点。
