甘蓝夜蛾CYP9A90基因的克隆及溴氰菊酯对其诱导表达

发布时间：2018-04-01 03:38

本文选题：甘蓝夜蛾　切入点：P　出处：《植物保护》2017年01期

【摘要】：P450 CYP9A家族基因与昆虫对拟除虫菊酯类杀虫剂的抗性相关。为证实溴氰菊酯对甘蓝夜蛾CYP9A基因的诱导效果,本研究采用RT-PCR和RACE技术克隆获得甘蓝夜蛾CYP9A基因,real-time PCR检测该基因在甘蓝夜蛾不同组织中的表达差异及溴氰菊酯处理甘蓝夜蛾5龄幼虫不同时间后该基因相对表达量变化,研究结果可为甘蓝夜蛾对溴氰菊酯的抗性治理提供依据。结果表明:克隆得到甘蓝夜蛾CYP9A基因cDNA全长序列,该序列包含1 828bp,包括1个125bp的5′非编码区,1个104bp的3′非编码区和1个1 599bp的开放阅读框,编码532个氨基酸,分子量约为61.1kDa,等电点为8.84,GenBank登录号为KR676343,被国际P450命名委员会命名为CYP9A90。Real-time PCR分析结果表明,该基因在甘蓝夜蛾5龄幼虫6个组织中表达情况不同,其中在脂肪体中表达量最高。低剂量溴氰菊酯作用后不同时间点CYP9A90mRNA总体呈现诱导表达趋势。
[Abstract]:P450 CYP9A family genes were associated with insect resistance to pyrethroid insecticides. The induction of deltamethrin on CYP9A gene of Spodoptera spp. In this study, RT-PCR and RACE techniques were used to clone the CYP9A gene of Spodoptera spp. The expression of CYP9A gene was detected by real-time PCR in different tissues of Spodoptera spp., and the relative expression level of the gene was changed after deltamethrin was treated with the 5th instar larvae of Spodoptera spp. The results may provide a basis for the management of resistance to deltamethrin. The results showed that the full-length cDNA sequence of CYP9A gene was obtained. The sequence consists of 1 828 BP, including 5 'non-coding region of 125bp, 3' noncoding region of 1 104bp and 1 open reading frame of 1 599bp, encoding 532 amino acids. The molecular weight is about 61.1 kDa, the isoelectric point is 8.84 GenBank accession number is KR676343.The results of CYP9A90.Real-time PCR analysis show that the gene is expressed differently in 6 tissues of 5th instar larvae of Spodoptera exigua. The expression level of CYP9A90mRNA was the highest in fat body, and the tendency of CYP9A90mRNA expression was induced at different time points after the treatment of low dose deltamethrin.
【作者单位】：东北农业大学农学院;东北农业大学资源与环境学院;
【基金】：黑龙江省教育厅科学技术研究项目(12531045)
【分类号】：S433

【相似文献】