JIA-2021-1984 脂肪酸结合蛋白基因在黄野螟蜕皮过程中的作用研究

doi:10.1016/j.jia.2022.08.003

摘要/Abstract

摘要：

【目的】脂肪酸结合蛋白（Fatty acid-binding protein）在细胞内以脂肪酸载体蛋白广泛参与脂肪酸的吸收、转运和代谢。它是昆虫脂质代谢中的关键蛋白，在昆虫的多种生理活动中发挥重要作用。研究黄野螟（Heortia vitessoides Moore）脂肪酸结合蛋白的时空表达特性及其在蜕皮发育中的生物学功能，为基于RNAi的黄野螟害虫防治技术提供理论基础。【方法】在黄野螟成虫转录组数据库中通过搜索关键词及在线比对获得黄野螟脂肪酸结合蛋白（HvFABP）的全序列，并对其进行生物信息学分析。以黄野螟不同龄期、不同的幼虫组织和成虫组织部位为材料提取RNA并反转录为cDNA，采用实时荧光定量PCR研究其时空表达特性；选取发育阶段相同的黄野螟幼虫进行饥饿胁迫、20E处理和RNA干扰探讨其生物学功能。【结果】本研究成功鉴定出一条黄野螟脂肪酸结合蛋白基因。同源性比对和系统发育树分析显示，HvFABP编码的氨基酸序列与鳞翅目昆虫的脂肪酸结合蛋白具有很高的相似性，在系统发育树上与家蚕的BmFABP1关系最密切。不同龄期和不同组织的实时荧光定量PCR结果表明HvFABP在各个龄期和组织中均有表达，其中在预蛹期到成虫期表达量较高，并在幼虫中肠和成虫翅膀的表达量最高。饥饿胁迫24h、48h时HvFABP表达被抑制，而在72h黄野螟进入预蛹期时表达量显著上调。20-hydroxyecdysone（20E）处理24h和48h时HvFABP表达量均上调，表明20E可诱导HvFABP的表达。RNA干扰显著沉默了HvFABP，导致黄野螟发育延迟、蜕皮异常和出现致死表型，存活率显著下降。【结论】这些结果表明HvFABP在黄野螟的蜕皮过程中起关键作用，该基因被沉默后黄野螟无法完成蜕皮而导致死亡。【创新性】昆虫脂肪酸结合蛋白基因功能多种多样，在基因功能的探索上远不如哺乳动物。本研究首次对黄野螟脂肪酸结合蛋白基因进行时空表达分析和基因功能研究，丰富昆虫脂肪酸结合蛋白基因功能的探索，为黄野螟生物防治靶标基因的合理选择提供理论基础。

Abstract:

As intracellular fatty acid (FA) carriers, FA-binding proteins (FABPs) widely participate in the absorption, transport, and metabolism of FAs. It is a key protein in insect lipid metabolism and plays an important role in various physiological activities of insects. An FABP gene (HvFABP) was cloned from the transcriptional library of Heortia vitessoides Moore (Lepidoptera: Crambidae), and its expression patterns were determined using reverse transcription quantitative PCR (RT-qPCR). Stage- and tissue-specific expression profiles indicated that HvFABP highly expressed from prepupal to adult stages and in larval midgut and adult wings. HvFABP expression may be induced through starvation, mRNA expression was downregulated at 24 and 48 h and upregulated at 72 h after starvation. Furthermore, 20-hydroxyecdysone can induce the upregulation of its expression. RNA interference-mediated silencing of HvFABP significantly inhibited HvFABP expression, resulting in delayed development, abnormal molting or lethal phenotypes, and a significantly reduced survival rate. These results indicate that HvFABP plays a key role in the molting of H. vitessoides

Key words: fatty acid-binding protein , starvation , 20-hydroxyecdysone , RNA interference , Heortia vitessoides Moore

. JIA-2021-1984 脂肪酸结合蛋白基因在黄野螟蜕皮过程中的作用研究[J]. Journal of Integrative Agriculture, 2023, 22(2): 495-504.

YE Qing-ya, LI Zhi-xing, CHEN Qing-ling, SUN Ming-xu, YIN Ming-liang, LIN Tong. Fatty acid-binding protein gene is indispensable for molting process in Heortia vitessoides (Lepidoptera: Crambidae)[J]. Journal of Integrative Agriculture, 2023, 22(2): 495-504.

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