Molecular cloning and characterization of a ryanodine receptor gene in brown planthopper (BPH), Nilaparvata lugens (Stal)

摘要

BACKGROUND: Ryanodine receptors (RyRs) are a distinct class of intracellular calcium (Ca~(2+)) release channel. The recent discovery of diamide insecticides has prompted studies on insect RyRs. However, information about the structure and function of insect RyRs is still limited. In this study, we isolated and characterized a full-length RyR cDNA (named NIRyR) from the brown planthopper, Nilaparvata lugens (Stal) (Homoptera: Delphacidae), a serious rice pest throughout Asia. RESULTS: The composite NIRyR gene containsan open reading frame of 15 423 bp encoding a protein of 5140 amino acid residues, which shares high sequence identity (78-81%) with other insect homologues, except for two regions (IDR1:4379-4732; IDR2: 1307-1529) with markedly low identity (44-48 and 38-41%, respectively). All hallmarks of the RyR proteins are conserved in the NIRyR protein, including the RyR domain as well as mannosyltransferase, IP3R and RyR (pfam02815) (MIR) and RyR and IP3R homology (pfamOl 365) (RIH) domains. Expression analysis of NIRyR revealed significant differences in mRNA expression levels among N. lugens developmental stages. Furthermore, three alternative splicing sites were identified in NIRyR, one of which forms the mutually exclusive exons A/B and is conserved in various insect species. Diagnostic PCR assays showed that the splice variant containing exon A was predominantly detected in all developmental stages. CONCLUSION: NIRyR may play an important role in the control of developmental processes of N. lugens. Alternative splicing may generate the functional diversity of NIRyR. The results provided the basis for further structural and functional characterization of NIRyR.