甲硫氨酸富集蛋白在棉铃虫卵巢发育中的功能及机制解析  

Functional mechanism of methionine-rich storage protein in the ovarian development of Helicoverpa armigera

作　　者：秦千越 方斌 王靖康 张博 耿梓宸 安世恒[1] 李祥 赵文丽 QIN Qianyue;FANG Bin;WANG Jingkang;ZHANG Bo;GENG Zichen;AN Shiheng;LI Xiang;ZHAO Wenli(College of Plant Protection,Henan Agricultural University,Zhengzhou 450046,China)

机构地区：[1]河南农业大学植物保护学院,河南郑州450046

出　　处：《河南农业大学学报》2025年第2期285-294,共10页Journal of Henan Agricultural University

基　　金：国家自然科学基金项目(31801339,32302345);河南省自然科学基金青年项目(232300420188);河南省高等学校重点科研项目(23A210005)。

摘　　要：【目的】探讨甲硫氨酸富集蛋白(methionine-rich storage protein, HaSP1)在棉铃虫(Helicoverpa armigera)卵巢发育中的功能及相关机制。【方法】分析蛋白质不足处理棉铃虫4龄幼虫对卵巢发育、HaSP1基因转录表达和JH滴度的影响;PCR扩增棉铃虫HaSP1基因开放阅读框序列,并进行生物信息学分析;利用实时荧光定量聚合酶链式反应(real-time quantitative polymerase chain reaction,qRT-PCR)检测HaSP1基因在棉铃虫羽化前3 d至羽化后4 d雌成虫卵巢和脂肪体中的转录表达变化;最后通过RNA干扰技术(RNA interference, RNAi)敲低雌成虫体内HaSP1基因的表达量,分析对卵巢发育及其体内总氨基酸含量、甲硫氨酸(methionine,Met)含量以及HaVg转录表达量的影响。【结果】棉铃虫幼虫期饲料中蛋白质不足,会显著抑制雌成虫的生殖能力,并导致JH滴度和HaSP1基因的转录显著降低。棉铃虫HaSP1基因的开放阅读框长2 256 bp,棉铃虫、黏虫(Mythimna loreyi)和粉纹夜蛾(Trichoplusia ni)中的SP1亲缘关系更近。HaSP1基因在卵巢中羽化前第2天高表达,在脂肪体中羽化前第3天高表达。敲低HaSP1基因转录会抑制棉铃虫卵巢发育,并显著降低HaVg转录、总氨基酸及Met含量。【结论】HaSP1通过影响甲硫氨酸含量从而在棉铃虫卵巢发育中发挥重要作用。【Objective】To investigate the function and mechanism of Methionine-rich storage protein(HaSP1)in the ovarian development of Helicoverpa armigera.【Method】The effects of protein shortage on ovarian development from four-instar larvae,HaSP1 transcriptional expression,and juvenile hormone(JH)titer were analyzed.The open reading frame of HaSP1 was amplified by PCR and subjected to bioinformatics analysis.The transcriptional expression profile of HaSP1 in the ovaries and fat bodies of females from three days before emergence to four days after emergence was measured using real-time quantitative polymerase chain reaction(qRT-PCR).Finally,the effects of HaSP1 knockdown in females’ovarian development,total amino acid content,methionine content,and HaVg transcriptional expression were analyzed using RNA interference(RNAi).【Result】Protein deficiency in the larval stage significantly inhibited the reproductive capacity of females and led to markedly decreases in JH titer and HaSP1 transcription.The open reading frame of HaSP1 is 2256 bp,and HaSP1 is more closely to SP1 in Mythimna loreyi,and Trichoplusia ni.HaSP1 was highly expressed in ovaries on the second day before emergence,and the expression peak in fat body occurred on the third day before emergence.Knocking down HaSP1 inhibited ovarian development and significantly reduced HaVg transcription,total amino acid content,and methionine content.【Conclusion】HaSP1 played an important role in the ovarian development of H.armigera by affecting methionine content.

关 键 词：棉铃虫 甲硫氨酸富集蛋白SP1 卵巢发育 RNA干扰 甲硫氨酸 

分 类 号：S433.4[农业科学—农业昆虫与害虫防治]