《环境昆虫学报》

本研究旨在明确棉铃虫Helicoverpa armigera（Hübner）几丁质合成酶1（HaCHS1）基因在不同发育时期和不同组织的表达及氟啶脲处理后对该基因表达的影响。利用RT-PCR和RACE技术克隆获得棉铃虫HaCHS1基因，HaCHS1基因全长为5 247 bp，其中开放阅读框4 698 bp，编码1 565个氨基酸，5′非翻译区为168 bp，3′非翻译区为381 bp。预测蛋白的相对分子质量为180.7 kDa，等电点为6.50，HaCHS1具有16个跨膜螺旋和6个N-糖基化位点。RT-qPCR检测结果表明，HaCHS1基因在预蛹期表达量最低，在蛹期第1天表达量最高。HaCHS1基因在头部的表达量最高，其次是表皮，而在中肠、气管、马氏管和脂肪体中的表达量均极低。亚致死浓度（60 mg/L）氟啶脲处理后，棉铃虫HaCHS1基因呈现出“抑制-激活-再抑制”的波动。本研究将为棉铃虫HaCHS1基因功能的研究奠定基础，为基于几丁质合成途径设计新药提供科学的理论依据。

英文摘要:

The objective of this study was to investigate spatio temporal expression profiles of chitin synthase 1 gene (HaCHS1) in Helicoverpa armigera (Hübner), and the effect of chlorfluazuron on the relative expression levels of HaCHS1. HaCHS1was cloned by reverse transcription PCR (RT-PCR) and rapid amplification cDNA ends (RACE) technologies. The length of HaCHS1 was 5 247 bp, with an open reading frame (ORF) of 4 698 bp, encoding 1 565 amino acids, a 5′untranslated region of 168 bp, and a 3′untranslated region of 381 bp. HaCHS1 had a theoretical molecular mass of 180.7 kDa, with an isoelectric point of 6.50, 16 transmembrane helices and 6 N-glycosylation sites.The results of real time quantitative PCR (RT-qPCR) assay showed that the relative expression levels of HaCHS1 were the lowest in prepupae stage and the highest on the first day of the pupae stage. In addition, the relative expression levelsof HaCHS1 were the highest in the head, followed by the epidermis, while very low in the midgut, trachea, malpighian tubules, and fatbody. After exposure to chlorfluazuron (60 mg/L), the relative expression levels of HaCHS1 exhibited “inhibition activation inhibition” fluctuations in H. armigera larvae. These results provide the favorable foundation for the study on the function of CHS1 gene inH. armigera, and provide a scientific theoretical basis for the design of novel drugs.

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