Molecular cloning, expression profiling and RNA interference of a vitellogenin gene from Harmonia axyridis(Coleoptera: Coccinellidae)  

作　　者：LIANG Chao LIU Ting-hui HAN Shi-peng HE Yun-zhuan 

机构地区：[1]College of Plant Protection, Hebei Agricultural University

出　　处：《Journal of Integrative Agriculture》2019年第10期2311-2320,共10页农业科学学报（英文版）

基　　金：supported by the National Natural Science Foundation of China (31272095);the National Key Research and Development Program of China (2017YFD0201004);the Hebei Modern Agriculture Industry Technology System, China (HBCT2018060204)

摘　　要：In this study, the Harmonia axyridis vitellogenin gene 2 (HmVg2) sequence was identified from transcriptomic databases of female adult H. axyridis and cloned into pMD18-T vector. The HmVg2 gene was 5 460 bp in length, and formed with an open reading frame (ORF) of 5 361 nucleotides (GenBank accession no. KY794939). The putative molecular weight of the primary HmVg2 protein was 203.459 kDa and the predicted isoelectric point (pI) was 8.59. HmVg2 contained a signal peptide, vitellogenin N-terminal (vitellogenin-N) domain, domain of unknown function 1943 (DUF1934) domain and von Willebrand factor type D (VWD) domain. The developmental expression profiling showed that HmVg2 was extremely highly expressed in female insects, but was expressed at lower levels in male insects. In female insects, HmVg2 was mainly expressed in the wing and fat body. The double-stranded RNA-HmVg2/-GFP was injected into H. axyridis, and qRT-PCR results showed that the HmVg2 gene was specifically silenced. The eggs laid during the first five days and the hatching rate of eggs was lower than controls after dsHmVg2 injection. This investigation demonstrated that the HmVg2 gene plays an important role in H. axyridis reproduction and enriches the function of the insect vitellogenin gene.In this study, the Harmonia axyridis vitellogenin gene 2(HmVg2) sequence was identified from transcriptomic databases of female adult H. axyridis and cloned into p MD18-T vector. The HmVg2 gene was 5 460 bp in length, and formed with an open reading frame(ORF) of 5 361 nucleotides(Gen Bank accession no. KY794939). The putative molecular weight of the primary HmVg2 protein was 203.459 k Da and the predicted isoelectric point(p I) was 8.59. HmVg2 contained a signal peptide, vitellogenin N-terminal(vitellogenin-N) domain, domain of unknown function 1943(DUF1934) domain and von Willebrand factor type D(VWD) domain. The developmental expression profiling showed that HmVg2 was extremely highly expressed in female insects, but was expressed at lower levels in male insects. In female insects, HmVg2 was mainly expressed in the wing and fat body. The double-stranded RNA-HmVg2/-GFP was injected into H. axyridis, and q RT-PCR results showed that the HmVg2 gene was specifically silenced. The eggs laid during the first five days and the hatching rate of eggs was lower than controls after ds HmVg2 injection. This investigation demonstrated that the HmVg2 gene plays an important role in H. axyridis reproduction and enriches the function of the insect vitellogenin gene.

关 键 词：VITELLOGENIN Harmonia axyridis expression profiling RNAi 

分 类 号：S[农业科学]