关键词: 棉铃虫')">棉铃虫, 离子型受体')">离子型受体, 表达谱')">表达谱, 荧光定量PCR')">荧光定量PCR, 原位杂交')">原位杂交

Abstract: 【Aim】 Ionotropic receptors (IRs) are newly discovered chemosensory receptors in insects, which play an important role in insects in sensing acids and amines in the environment. The studies of cloning, sequence alignment and expression localization of genes of IRs will help their preliminary functional analysis in Helicoverpa armigera. 【Methods】 Based on the results of antennal transcriptome analysis, the full-length sequence of an ionotropic receptor gene of H. armigera was cloned by PCR, and its sequence structure was analyzed. The relative expression levels of this gene in different tissues of female and male adults of H. armigera were analyzed by real-time fluorescent quantitative PCR assay, and its expression localization in male antennae was detected by in situ hybridization. 【Results】 The full-length cDNA sequence of HarmIR8agene (GenBank accession no.: MH638313) was cloned from H. armigera. The HarmIR8a sequence comprises a 2 688 bp ORF and encodes a protein of 895 amino acids with three predicted transmembrane domains. HarmIR8acontains one amino-terminal domain (ATD), one ligand binding domain (LBD) consisting of two lobes (S1 and S2), one pore loop (P), three transmembrane domains (M1, M2 and M3) and one C-terminus. The qPCR results revealed that HarmIR8awas mainly expressed in the head (with appendages removed) and antennae of female and male adults of H. armigera, with the highest relative expression level in the antennae. The results of in situ hybridization of the male adult antennae of H. armigera showed that HarmIR8awas mainly expressed in the antennal coeloconic sensilla. 【Conclusion】 The transcriptional level and expression localization of HarmIR8awere preliminarily determined. This study lays a foundation for further functional characterization and physiological activities of ionotropic receptor genes in H. armigera.

Key words: Helicoverpa armigera, ionotropic receptors, expression profile, -time fluorescent quantitative PCR')">real-time fluorescent quantitative PCR, in situ hybridization