白背飞虱两个P450基因的克隆、序列分析及杀虫剂胁迫下的表达分析

doi:10.16409/j.cnki.2095-039x.2019.06.014

中国生物防治学报 ›› 2020, Vol. 36 ›› Issue (1): 87-96.DOI: 10.16409/j.cnki.2095-039x.2019.06.014

白背飞虱两个P450基因的克隆、序列分析及杀虫剂胁迫下的表达分析

贾泽艳¹, 周操², 曾庆会¹, 石敏¹, 杨洪^2,3, 金道超²

1. 贵州大学农学院, 贵阳 550025;
2. 贵州大学昆虫研究所/贵州山地农业病虫害重点实验室, 贵阳 550025;
3. 贵州大学烟草学院, 贵阳 550025

收稿日期:2019-04-03 出版日期:2020-02-08 发布日期:2020-02-26
通讯作者: 杨洪,教授,E-mail:axyridis@163.com
作者简介:贾泽艳,E-mail:2077130768@qq.com
基金资助:
国家自然科学基金（31560522，31960537）；贵州省农业科技攻关项目（2013-3006）；贵州省农业科技攻关项目（2010-3064）；大学生创新创业训练计划（201810657023）

Cloning and Sequence Analysis of Two P450 Genes and Expression of the Genes in Sogatella furcifera under Insecticides Stress

JIA Zeyan¹, ZHOU Cao², ZENG Qinghui¹, SHI Min¹, YANG Hong^2,3, JIN Daochao²

1. Agricultural College of Guizhou University, Guiyang 550025, China;
2. Institute of Entomology, Guizhou University/Provincial Key Laboratory for Agricultural Pest Management of Mountainous Regions, Guiyang 550025, China;
3. College of Tobacco Science, Guizhou University, Guiyang 550025, China

Received:2019-04-03 Online:2020-02-08 Published:2020-02-26

摘要/Abstract

摘要： 细胞色素P450属于超基因家族，与昆虫对杀虫剂抗药性产生有重要关系。本文在白背飞虱转录组数据的基础上，克隆到了两个P450基因，分别命名为SfCYP4DE1和SfCYP353D1v2，其分别编码534和471个氨基酸，分子量约为61.567和53.762 kDa，理论等电点为9.18和8.74。结构域分析发现，SfCYP4DE1和SfCYP353D1v2氨基酸序列中均包含5个P450基因共有序列。噻虫嗪、噻嗪酮和阿维菌素的LC₁₀、LC₂₅、LC₅₀和LC₉₀4个不同浓度处理白背飞虱3龄若虫48 h后，采用RT-qPCR技术测定SfCYP4DE1和SfCYP353D1v2两个基因的相对表达量，结果表明：噻虫嗪、噻嗪酮LC₁₀和LC₂₅浓度胁迫后SfCYP4DE1基因相对表达量均显著低于对照组，但随着浓度继续的加大，SfCYP4DE1基因的表达量出现上升的趋势。不同的是，经阿维菌素LC₁₀和LC₂₅浓度胁迫后，SfCYP4DE1基因的表达量均显著高于对照组，且随着浓度增加其表达量升高。与对照组相比，经噻虫嗪不同浓度胁迫后，SfCYP353D1v2基因表达量均呈现出诱导增加的趋势，且LC₁₀、LC₅₀和LC₉₀处理组显著高于对照组。噻嗪酮LC₁₀浓度处理能够显著诱导SfCYP353D1v2基因的表达，但阿维菌素LC₁₀浓度处理下其表达量被显著抑制。另外，以阿维菌素LC₅₀和LC₉₀浓度处理后，SfCYP353D1v2基因的表达量被显著诱导。以上研究结果可为进一步研究白背飞虱P450基因对杀虫剂解毒代谢途径提供基础资料。

关键词: 白背飞虱, 细胞色素P450, 基因克隆, 杀虫剂

Abstract: Cytochrome P450 is a supergene family, which is closely related to insecticides resistance. Based on the transcriptome data of Sogatella furcifera, two P450 genes, SfCYP4DE1 and SfCYP353D1v2, were cloned. SfCYP4DE1 and SfCYP353D1v2 encode 534 and 471 amino acids with molecular weight of 61.567 and 53.762 kDa and theoretical isoelectric point of 9.18 and 8.74, respectively. Domain analysis showed that the amino acid sequences of SfCYP4DE1 and SfCYP353D1v2 contained the common sequences of 5 P450 genes. The relative expression levels of SfCYP4DE1 and SfCYP353D1v2 were determined by RT-qPCR at 48h post treatment of 3^rd instar S. furcifera with different concentrations (LC₁₀, LC₂₅, LC₅₀, and LC₉₀) of thiamethoxam, buprofezin and avermectin. The relative expression of SfCYP4DE1 gene was significantly lower in the insects treated with thiamethoxam and buprofezin at both LC₁₀ and LC₂₅ than that in the control insects. However, with the further increase of insecticide concentration, the expression of SfCYP4DE1 gene showed an increasing trend. In contrast, when the insects were treated with avermectin at LC₁₀ and LC₂₅, the expression of SfCYP4DE1 gene was significantly higher than that in the control insects; and the expression level increased with the increase of avermectin concentration. For the SfCYP353D1v2 gene, expression was induced by different concentrations of thiamethoxam in comparison with the control, and was significantly higher in the LC₁₀, LC₅₀ and LC₉₀ treatments than in the control. Treatment with buprofezin at LC₁₀ significantly induced expression of the SfCYP353D1v2 gene, while treatment with avermectin at LC₁₀ significantly inhibited the gene expression. However, treatment with avermectin at LC₅₀ and LC₉₀ significantly induced expression of the SfCYP353D1v2 gene. These results provide basic information for further study of the roles of P450 genes in detoxification and metabolism of insecticides in S. furcifera.

Key words: Sogatella furcifera, cytochrome P450, gene cloning, insecticide

中图分类号:

S435.112

贾泽艳, 周操, 曾庆会, 石敏, 杨洪, 金道超. 白背飞虱两个P450基因的克隆、序列分析及杀虫剂胁迫下的表达分析[J]. 中国生物防治学报, 2020, 36(1): 87-96.

JIA Zeyan, ZHOU Cao, ZENG Qinghui, SHI Min, YANG Hong, JIN Daochao. Cloning and Sequence Analysis of Two P450 Genes and Expression of the Genes in Sogatella furcifera under Insecticides Stress[J]. Chinese Journal Of Biological Control, 2020, 36(1): 87-96.

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白背飞虱两个P450基因的克隆、序列分析及杀虫剂胁迫下的表达分析

Cloning and Sequence Analysis of Two P450 Genes and Expression of the Genes in Sogatella furcifera under Insecticides Stress

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