Cloning And Tissue Expression Of Odorant Binding Proteins Gene In Callosobruchus Chinensis(coleoptera:Bruchidae)

Callosobruchus chinensis is a dominant pest in the seeds of leguminous plants such as mung beans,cowpeas,and red beans.It has a strong infection ability and hidden life history,which poses a serious threat to legume storage.Odorant binding proteins(OBPs)play an important role in the process of insect olfactory recognition.The study of C.chinensis OBPs is helpful to understand its olfactory perception mechanism and lay the molecular foundation for the development of prevention and control technology based on olfactory regulation.This study is based on C.chinensis antennal transcriptome data.The main results are as follows:(1)6 CchiOBPs genes(CchiOBP1 ~ CchiOBP6)of C.chinensis were cloned,and the open reading frame sizes of Cchi OBP1,Cchi OBP2,Cchi OBP3,Cchi OBP4,Cchi OBP5 and Cchi OBP6 were 423,432,396,441,279,and 405 bp.Cchi OBP1,Cchi OBP2,Cchi OBP3,Cchi OBP4,and Cchi OBP6 are Classcial OBPs,Cchi OBP5 is an incomplete Minus-C OBP.Using software to predict that all 6 Cchi OBPs contain signal peptides,and Cchi OBP2 has relatively strong hydrophilicity.Phylogenetic analysis showed that Cchi OBP1,Cchi OBP2,and Cchi OBP5 had high homology with OBPs of Chrysomelidae,and Cchi OBP3,Cchi OBP4,and Cchi OBP6 had high homology with OBPs of the Cerambycidae.(2)The main fragments of 8 candidate internal reference genes were amplified,and the best internal reference gene were determined by q RT-PCR.By analyzing the Ct value,the evaluation results of the 3softwares of ge Norm,Norm Finder and Best Keeper are integrated,β-Tubulin was identified as the best internal reference gene in different tissues of adult C.chinensis.(3)Analysis of the expression levels of different tissues of adult C.chinensis showed that 6Cchi OBPs were expressed in different amounts in the antennae,heads,abdomen,wings and feet.Cchi OBP1,Cchi OBP2,Cchi OBP3,Cchi OBP4,and Cchi OBP6 all showed high expression in male and female C.chinensis antennae,and were significantly higher than the other 4 tissues.Cchi OBP5 is relatively special,showing high expression in female antennae and heads;but in males,the expression level of the feet is significantly higher than other tissues,and the expression level in antennae is the lowest,which may be related to its function.