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星豹蛛PaCYP3001U16基因的克隆与表达分析
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引用本文:王雅丽,赵瑞,王美,张晓晨,燕晶晶,李锐.星豹蛛PaCYP3001U16基因的克隆与表达分析.植物保护学报,2022,49(2):603-611
DOI:10.13802/j.cnki.zwbhxb.2022.2021153
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作者单位E-mail
王雅丽 山西农业大学植物保护学院, 太谷 030801  
赵瑞 山西农业大学植物保护学院, 太谷 030801  
王美 山西农业大学植物保护学院, 太谷 030801  
张晓晨 山西农业大学植物保护学院, 太谷 030801  
燕晶晶 山西农业大学植物保护学院, 太谷 030801  
李锐 山西农业大学植物保护学院, 太谷 030801 sxaulr@126.com 
中文摘要:为探究细胞色素P450基因在星豹蛛Pardosa astrigera体内的表达特性及对溴氰菊酯胁迫的响应,采用反转录PCR技术克隆其细胞色素P450基因并进行序列分析,采用实时荧光定量PCR (quantitative real-time PCR,RT-qPCR)技术分析其在星豹蛛各发育阶段及雌雄成蛛不同部位的表达水平,同时分析其在溴氰菊酯不同浓度胁迫下和不同处理时间后的表达模式。结果显示,在星豹蛛雌成蛛中克隆得到1个细胞色素P450基因,其开放阅读框为1 473 bp,编码490个氨基酸,命名为PaCYP3001U16(GenBank登录号为MZ643213)。PaCYP3001U16基因在星豹蛛各发育阶段均有表达,其中在成蛛期的表达量最高,在6龄幼蛛期的表达量最低;该基因在雌雄成蛛腹部的表达量显著高于在头胸部及足部的表达量。星豹蛛雄成蛛经LC10(5.151 mg/L)、LC30(8.619 mg/L)和LC50(12.311 mg/L)溴氰菊酯处理12 h,PaCYP3001U16基因的表达量均被抑制,且在LC50处理下表达量最低。PaCYP3001U16基因经LC30溴氰菊酯处理不同时间后表达趋势不同,在处理2 h和4 h表现为诱导效应,且在处理4 h的基因表达量最高,为对照的1.79倍;在处理8、12和24 h表现为抑制效应,在处理24 h的基因表达量最低;而处理48 h的基因表达量又开始升高,表现为诱导效应。表明星豹蛛PaCYP3001U16基因在不同发育阶段和成蛛不同部位的表达水平不同,且溴氰菊酯可以诱导该基因的表达,推测该基因参与星豹蛛的生长发育及对外源物质的代谢。
中文关键词:星豹蛛  细胞色素P450  基因克隆  序列分析  溴氰菊酯  表达分析
 
Cloning and expression analysis of cytochrome P450 gene PaCYP3001U16 in a wolf spider Pardosa astrigera
Author NameAffiliationE-mail
WANG Ya-li College of Plant Protection, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China  
ZHAO Rui College of Plant Protection, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China  
WANG Mei College of Plant Protection, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China  
ZHANG Xiao-chen College of Plant Protection, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China  
YAN Jing-jing College of Plant Protection, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China  
LI Rui College of Plant Protection, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China sxaulr@126.com 
Abstract:In order to explore the expression characteristics of cytochrome P450 gene in a wolf spider Pardosa astrigera and its response to deltamethrin stress, reverse transcription-PCR was used to clone the gene and its sequence was analyzed. Quantitative real-time PCR(RT-qPCR) was used to analyze the expression levels of cytochrome P450 gene at different developmental stages and in different body parts of female and male adults of P. astrigera. The expression pattern of cytochrome P450 gene in P. astrigera was analyzed under different concentrations of deltamethrin and different treatment durations. The results showed that a cytochrome P450 gene was cloned from female adults of P. astrigera with an open reading frame of 1 473 bp, encoding 490 amino acids, named PaCYP3001U16(GenBank accession no.MZ643213). The PaCYP3001U16 gene was expressed at all developmental stages of P. astrigera; its expression was the highest at the adult stage and the lowest at the 6th-instar larval stage, and the expression in the abdomen of female and male adults of P. astrigera was significantly higher than those in cephalothorax and legs. Exposure of male adults of P. astrigera to deltamethrin at three concentrations(LC10: 5.151 mg/L, LC30: 8.619 mg/L and LC50: 12.311 mg/L) for 12 h led to the inhibition of the expression of PaCYP3001U16 gene, and the lowest expression was found in LC50treatment. The expression levels of PaCYP3001U16 gene were different after LC30deltamethrin treatment for different periods:the gene expression exhibited an induction effect at 2 h and 4 h, and the highest expression was detected at 4 h, 1.79 times of that in the control. The expression of PaCYP3001U16 gene displayed an inhibition effect at 8, 12 and 24 h, and the lowest expression occurred at 24 h. Its expression rebounded at 48 h,showing an induction effect. The PaCYP3001U16 gene had different expression levels at different developmental stages and in different body parts of adults of P. astrigera. Deltamethrin could induce the expression of PaCYP3001U16 gene. It indicated that PaCYP3001U16 gene was involved in the growth and development of P. astrigera and metabolism of exogenous substances.
keywords:Pardosa astrigera  cytochrome P450  gene cloning  sequence analysis  deltamethrin  expression analysis
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