Cloning and spatiotemporal expression profiling of the 14-3-3 genes in Bemisia tabaci MED(Hemiptera:Aleyrodidae)
[Aim]14-3-3 proteins are a class of regulatory proteins found in eukaryotic organisms,which can be involved in signal transduction,immune response,growth and development,and stress response.The aim of this study is to clone the full-length cDNA sequences of the 14-3-3 genes in Bemisia tabaci MED,and understand the characteristics of the proteins encoded by 14-3-3 genes and the spatiotemporal expression patterns of 14-3-3 genes.[Methods]The full-length cDNA sequences of 14-3-3 genes of B.tabaci MED were cloned by RT-PCR,and their biological properties were analyzed by bioinformatics software and online website.RT-qPCR was used to determine the expression levels of 14-3-3 genes in different developmental stages(egg,1st-4th instar nymphs and adult),in adult male and female,and in the head,thorax and abdomen of female adult of B.tabaci MED.[Results]Two subtypes of the 14-3-3 gene of B.tabaci MED were cloned and characterized:Bt14-3-3 epsilon(GenBank accession no.:XM_019046102.1)and Bt14-3-3 zeta(GenBank accession no.:XM_019057395.1).The open reading frames(ORFs)of Bt14-3-3 epsilon and Bt14-3-3 zeta were 771 and 744 bp,encoding 256 and 247 amino acids,respectively.The proteins encoded by Bt14-3-3 epsilon and Bt14-3-3 zeta were hydrophilic proteins without transmembrane helical region and signal peptide,and their secondary structure mainly consisted of α-helices.Phylogenetic tree analysis showed that Bt14-3-3 epsilon was clustered into one cluster with 14-3-3 epsilon proteins of Nilaparvata lugens,Cimex lectularius and Halyomorpha halys,sharing higher homology,while Bt14-3-3 zeta was more closely related to 14-3-3 zeta of N.lugens.RT-qPCR results showed that Bt14-3-3 epsilon and Bt14-3-3 zeta had higher expression levels in the egg,female adults and abdomen of female adults of B.tabaci MED.[Conclusion]The full-length sequence,characteristics of the coded proteins and spatiotemporal expression of two subtypes of the 14-3-3 gene of B.tabaci MED have been clarified.The results of this study provide a basis for subsequent studies on the molecular function of 14-3-3 proteins.
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