Ryanodine receptor genes of the rice stem borer, Chilo suppressalis: Molecular cloning, alternative splicing and expression profiling

Pestic Biochem Physiol. 2017 Jan:135:69-77. doi: 10.1016/j.pestbp.2016.06.002. Epub 2016 Jun 8.

Abstract

The ryanodine receptor (RyR) of the calcium release channel is the main target of anthranilic and phthalic diamide insecticides which have high selective insecticidal activity relative to mammalian toxicity. In this study, the full-length cDNA of Chilo suppressalis RyR (CsRyR) was isolated and characterized. The CsRyR mRNA has an open reading frame (ORF) of 15,387bp nucleotides, which encodes 5128 amino acids with GenBank ID: KR088972. Comparison of protein sequences showed that CsRyR shared high identities with other insects of 77-96% and lower identity to mammals and nematodes with only 42-45%. One alternative splicing site (KENLG) unique to Lepidoptera was found and two exclusive exons of CsRyR (I /II) were revealed. Spatial and temporal expression of CsRyR mRNA was at the highest relative level in 3rd instar larvae and head (including brain and muscle), and at the lowest expression level in egg and fat body. The expression levels of whole body CsRyR mRNA were increased remarkably after injection of 4th instar larvae with chlorantraniliprole at 0.004 to 0.4μg/g. This structural and functional information on CsRyR provides the basis for further understanding the selective action of chlorantraniliprole and possibly other diamide insecticides.

Keywords: Alternative splicing; Chilo suppressalis; Chlorantraniliprole, phylogenetic relationships; Ryanodine receptor; mRNA expression.

MeSH terms

  • Alternative Splicing
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • Gene Expression Profiling
  • Insect Proteins / genetics*
  • Insecticides / toxicity*
  • Larva / drug effects
  • Larva / genetics*
  • Lepidoptera / drug effects
  • Lepidoptera / genetics*
  • Phylogeny
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Ryanodine Receptor Calcium Release Channel / genetics*
  • Sequence Analysis, DNA
  • ortho-Aminobenzoates / toxicity*

Substances

  • DNA, Complementary
  • Insect Proteins
  • Insecticides
  • RNA, Messenger
  • Ryanodine Receptor Calcium Release Channel
  • ortho-Aminobenzoates
  • chlorantranilipole