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Chinese Journal of Biological Control ›› 2021, Vol. 37 ›› Issue (3): 486-494.DOI: 10.16409/j.cnki.2095-039x.2021.01.015

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Ligand Binding Characteristics of the Chemosensory Protein AipsCSP2 from Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae)

RAO Fuqiang1,2, SU Xu1,2, LI Zibo2, GENG Ting3, ZHANG Yongjun2, SONG Ping1, GU Shaohua4   

  1. 1. College of Plant Protection, Agricultural University of Hebei, Baoding 071000, China;
    2. State Key Laboratory for Biology of Plant Diseases and Insect Pests/Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
    3. Langfang Experimental Station of Crop Pests Science, MOA, Langfang 065000, China;
    4. College of Plant Protection, China Agricultural University, Beijing 100193, China
  • Received:2020-05-27 Online:2021-06-08 Published:2021-02-23

Abstract: To clarify the expression of AipsCSP2 gene in male and female adult tissues of Agrotis ipsilon (Hufnagel), we investigate the ligand binding characteristics of the AipsCSP2 and explore its functions. According to the pheromone gland transcriptome data, the AipsCSP2 gene was cloned and analyzed by bioinformatics and phylogenetic analysis. The expression level of the gene in different tissues of male and female adult was determined by qPCR. The recombinant protein was expressed and purified with prokaryotic expression system. Finally, the binding ability of this protein to the sex pheromone components of A. ipsilon and other Lepidoptera insects, as well as the volatiles of the main host plants were determined by fluorescence competitive binding assay. Results showed that the open reading frame (ORF) of AipsCSP2 gene was 360 bp and it encoded 119 amino acids. It has four conserved cysteines in the amino acid sequence and is a typical chemosensory protein. The tissue expression patterns showed that it was specifically high expressed in the female adult pheromone gland and male adult accessory gland, low expressed in other tissues. Fluorescent competitive binding assay results showed that AipsCSP2 has a very strong binding ability to cis-11-hexadecylaldehyde, the precursor of A. ipsilon sex pheromone cis-11-hexadecylacetate, with Ki of 1.48 μmol/L. It also has a strong binding ability to alcoholic and aldehydic sex pheromone of other Lepidoptera insects, such as tetradecanol, cis-9-hexadecaldehyde and cis-7-hexadecaldehyde, with Ki of 1.10 μmol/L, 0.63 μmol/L and 1.51 μmol/L. It has a moderate ability to the sex pheromone components of A. ipsilon cis-7-dodecanoacetate and cis-8-dodecanoacetate. The binding ability to the volatiles of the main host plants of A. ipsilon was weak or not. It is suggested that AipsCSP2 may be involved in the synthesis and recognition of sex pheromone in A. ipsilon, and play an important role in reproductive activities.

Key words: Agrotis ipsilon, chemosensory protein, AipsCSP2, tissue expression, prokaryotic expression, fluorescent competitive binding assay

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