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Chinese Journal of Biological Control ›› 2021, Vol. 37 ›› Issue (3): 495-507.DOI: 10.16409/j.cnki.2095-039x.2021.03.038

• RESEARCH REPORTS • Previous Articles    

Cloning, Sequence Analysis and Expression Characteristics of FoccOBP3 in Western Flower Thrips Frankliniella occidentalis

ZHANG Zhike1, HU Hua2, MA Rong3   

  1. 1. Ningxia Key Laboratory of Plant Diseases and Pests Control, Institute of Plant Protection, Ningxia Academy of Agriculture and Forestry Sciences, Yinchuan 750002, China;
    2. College of Agriculture, Ningxia University, Yinchuan 750021, China;
    3. Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2020-03-11 Published:2021-06-16

Abstract: Identifying the olfactory relative proteins in insects and studying their sequence and temporal-spatial expression will lay a basis for throwing light on the chemical communication mechanism between insect and their host plant. A new OBP gene in F. occidentalis was cloned, identified and named as FoccOBP3. The number in GenBank is MT682352. The full length of FoccOBP3 cDNA is 1226 bp. The open reading frame is 432 bp encoding a putative protein of 143 amino acids. The arrangement of typical six-cysteine is C1−X26−C2−X3−C3−X37−C4− X10−C5−X8−C6, which completely conforms to the typical OBPs' structural model about six conservative cysteine sites. The deduced amino acid sequence contains 5 lipotropism areas. A significant lipotropy pocket is formed by 62-75 amino acid residues. The molecular mass and isoelectric point of this amino acid sequence are 15.50 kD and 5.24, respectively. There was a putative signal peptide consist of 21 amino acid residues at the N terminus. The amino acid sequence of FoccOBP3 has the highest homolog with Diachasma alloeum DallGOBP (GenBank number is XP_015125081.1), Tribolium castaneum TcasPBP (GenBank number is XP_015835846.1) and TcasOBP07 (GenBank number is EFA04593.1) FoccOBP3 amino acid sequence is clusted into one small group with the amino acid sequence of Nicrophorus vespilloides NvesGOBP (GenBank number is XP_017781594.1), suggesting that these genes likely develop from a common ancestral gene. FoccOBP3 protein contains 127 amino acid residues that make up α−helix, 58 amino acid residues that form β−fold and 15 amino acid residues that form β−corner. The skeleton of FoccOBP3 protein is composed of six α−helices and the fold back connecting these helices, of which five α−helices form a binding pocket. The result of FoccOBP3 expression in different development stages of F. occidentalis indicated that this gene expresses highly in the first instar nymph. The next is in 5−day−old adult female. The expression levels of other development stages are lower very much. In addition, there are also difference in the expression between males and females. The expressions of female on day 1, 5 and 15 are higher than those of male at the same developmental stage except on day 10. The result of FoccOBP3 expression in different tissues of F. occidentalis revealed this gene express highly in the antennae. The next is in head. The expression levels of abdomen and leg are lower. The expression level of thorax is the lowest. An odorant binding protein FoccOBP3 from F. occidentalis was cloned in this study. The sequence characteristics of nucleotides and amino acids of FoccOBP3 were clarified. The secondary structure and three-dimensional model of FoccOBP3 were predicted successfully. Temporal-spatial expression pattern of FoccOBP3 was determined, suggesting that FoccOBP3 may play a significant role in odor recognition and host location of F. occidentalis.

Key words: Frankliniella occidentalis, odorant binding protein, molecular cloning, sequence analysis, structure prediction, spatial-temporal expression

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