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草地贪夜蛾卵黄原蛋白基因的分子特征及表达特性
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引用本文:赵静,邱之雨,肖留斌,谭永安,姜义平,柏立新.草地贪夜蛾卵黄原蛋白基因的分子特征及表达特性.植物保护学报,2021,48(4):798-805
DOI:10.13802/j.cnki.zwbhxb.2021.2020268
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作者单位E-mail
赵静 江苏省农业科学院植物保护研究所, 南京 210014  
邱之雨 扬州大学园艺与植物保护学院, 江苏 扬州 225009  
肖留斌 江苏省农业科学院植物保护研究所, 南京 210014 xlbwll@sohu.com 
谭永安 江苏省农业科学院植物保护研究所, 南京 210014  
姜义平 江苏省农业科学院植物保护研究所, 南京 210014  
柏立新 江苏省农业科学院植物保护研究所, 南京 210014  
中文摘要:为解析草地贪夜蛾Spodoptera frugiperda生殖调控的分子机制,利用反转录-聚合酶链式扩增(reverse transcription-polymerase chain reaction,RT-PCR)及cDNA末端快速扩增(rapid amplificationof cDNAends,RACE)技术克隆草地贪夜蛾生殖关键基因卵黄原蛋白(vitellogenin,Vg)全长序列,通过实时荧光定量PCR检测其Vg基因在不同性别、发育阶段及组织中的表达水平,并利用体视镜解剖观察不同日龄雌成虫的卵巢发育进度,分析其与Vg基因表达的相关性。结果显示,草地贪夜蛾Vg基因的cDNA序列全长为5 727 bp,开放阅读框长度为5 250 bp,编码1 749个氨基酸,预测编码蛋白的分子量为199.23 kD,N端前15个氨基酸为信号肽。基于鳞翅目昆虫Vg构建的系统发育树显示草地贪夜蛾Vg与斜纹夜蛾S.litura的Vg亲缘关系最近。草地贪夜蛾Vg基因在雌成虫脂肪体中特异性高表达,在雄成虫及雌成虫卵巢和表皮等其他组织中微表达;Vg在幼虫和化蛹初期表达量极低,从化蛹第8天起微弱表达,进入成虫期后表达量迅速上升,于羽化第4天雌成虫中达到表达峰值,随后降低。草地贪夜蛾Vg在卵巢的乳白透明期和卵黄沉积期上调表达,在成熟待产期达到峰值,进入产卵盛期及末期后表达量快速下降,表明草地贪夜蛾Vg在雌成虫期的表达动态与其卵巢发育进度紧密相关。
中文关键词:草地贪夜蛾  卵黄原蛋白  基因克隆  时空表达
 
Molecular and expression characteristics of vitellogenin gene in fall armyworm Spodoptera frugiperda
Author NameAffiliationE-mail
Zhao Jing Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, Jiangsu Province, China  
Qiu Zhiyu College of Horticulture and Plant Protection, Yangzhou University, Yangzhou 225009, Jiangsu Province, China  
Xiao Liubin Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, Jiangsu Province, China xlbwll@sohu.com 
Tan Yong'an Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, Jiangsu Province, China  
Jiang Yiping Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, Jiangsu Province, China  
Bai Lixin Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, Jiangsu Province, China  
Abstract:To analyze the molecular mechanism of reproductive regulation of fall armyworm Spodoptera frugiperda, the full-length sequence of S. frugiperda vitellogenin (Vg), a key gene for reproduction, was cloned by using reverse transcription-PCR (RT-PCR) and rapid amplification of cDNAends (RACE) techniques. The Vg expression profiles at different genders, tissues and developmental stages of S. frugiperda were studied by quantitative real-time PCR (qPCR); the ovaries of female adults at different days after eclosion were dissected by stereoscope to observe ovary development progress, and their correlation with Vg expression were analyzed. The results showed that S. frugiperda Vg cDNA sequence was 5 727 bp in length. The open read frame (ORF) of Vg was 5 250 bp, which encoded 1 749 amino acids. The predicted molecular weight of the encoded protein was 199.23 kD, and the first 15 amino acids at the N-terminus were signal peptides. Phylogenetic tree based on Lepidoptera Vgs showed that S. frugiperda Vg was closely related to the Vg of S. litura. The qPCR results showed that Vg was specifically highly expressed in the fat bodies of female adults and weakly expressed in males and other female adult tissues such as ovaries and epidermis. The Vg expression level was extremely low in larval and early pupal stage. Vg was weakly expressed from the 8th day of pupae, increased rapidly after entering the adult stage, reached a peak in the four-day-old female adult, and then decreased. The expression level of Vg gene increased rapidly at the opalescent transparent stage and yolk deposition stage of S. frugiperda ovaries, reached the peak in the egg maturation stage, and decreased rapidly after entering peak phase and terminal phase of oviposition. The results indicated that the expression dynamics of Vg in S. frugiperda female adults was closely related to ovarian development progress.
keywords:Spodoptera frugiperda  vitellogenin  gene cloning  spatiotemporal expression
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