Identification and Screening of LITAF Family Key Genes Responsive to Plant Secondary Metabolites in Helicoverpa armigera

棉铃虫中响应植物次生代谢物的LITAF家族关键基因的鉴定与筛选

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Abstract

Lipopolysaccharide-induced tumor necrosis factor alpha factor (LITAF) as a crucial cytokine can mediate the inflammatory and immune regulatory responses of an organism. It plays a significant mediatory role in the innate immune system of insects. Firstly, the LITAF family of cotton bollworm (Helicoverpa armigera, Lepidoptera: Noctuidae) was analyzed, and the differentially expressed genes of HaLITAFs were screened from the 2-tridecanone-treated transcriptome. Subsequently, these two differentially expressed HaLITAF genes were cloned and analyzed, and the effect of different larval stages, tissues, and plant secondary metabolites on their gene expression were detected in H. armigera. The cotton bollworm contains 13 LITAF genes, and there are no repetitive sequences among them. Upon 2-tridecanone treatment, only HaLITAF5 and HaLITAF7 were significantly upregulated in the LITAF family of cotton bollworm larvae after 6 h. The HaLITAF5 and HaLITAF7 proteins, respectively, comprised 78 and 113 amino acids, and both contained a CXXC motif, hydrophobic amino acid region and HXCPXCXXXXG motif. Both of them belong to the LITAF analogues (zf-LITAF-like) superfamily. The expressions of HaLITAF5 and HaLITAF7 were both the lowest in fourth-instar larvae and the highest in sixth-instar larvae, and both were abundantly expressed in the midgut of sixth-instar larvae. Following a 28 h treatment with 2-tridecanone, the expression levels of HaLITAF5 and HaLITAF7 peaked. HaLITAF7 exhibited the highest expression level after a 4 h exposure to tannic acid, whereas HaLITAF5 reached its peak after 28 h of tannic acid treatment. The maximum expression levels of HaLITAF5 and HaLITAF7 were observed after 28 h and 4 h of quercetin treatment, respectively. Subsequent to ZQ-8 treatment, HaLITAF5 expression peaked at 28 h, while HaLITAF7 expression peaked at 8 h. Both HaLITAF5 and HaLITAF7 were overexpressed after different kinds of plant secondary metabolite stresses, and thereby, they may be regulated in the expression of genes related to downstream detoxification metabolic processes. This provides a theoretical foundation for further studies on the functional mechanism of the LITAF gene in H. armigera.

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