Cloning, Characterization, and RNA Interference Effect of the UDP-N-Acetylglucosamine Pyrophosphorylase Gene in Cnaphalocrocis medinalis

Genes (Basel). 2021 Mar 24;12(4):464. doi: 10.3390/genes12040464.

Abstract

The rice leaf folder, Cnaphalocrocis medinalis is a major pest of rice and is difficult to control. UDP-N-acetylglucosamine pyrophosphorylase (UAP) is a key enzyme in the chitin synthesis pathway in insects. In this study, the UAP gene from C. medinalis (CmUAP) was cloned and characterized. The cDNA of CmUAP is 1788 bp in length, containing an open reading frame of 1464 nucleotides that encodes 487 amino acids. Homology and phylogenetic analyses of the predicted protein indicated that CmUAP shared 91.79%, 87.89%, and 82.75% identities with UAPs of Glyphodes pyloalis, Ostrinia furnacalis, and Heortia vitessoides, respectively. Expression pattern analyses by droplet digital PCR demonstrated that CmUAP was expressed at all developmental stages and in 12 tissues of C. medinalis adults. Silencing of CmUAP by injection of double-stranded RNA specific to CmUAP caused death, slow growth, reduced feeding and excretion, and weight loss in C. medinalis larvae; meanwhile, severe developmental disorders were observed. The findings suggest that CmUAP is essential for the growth and development of C. medinalis, and that targeting the CmUAP gene through RNAi technology can be used for biological control of this insect.

Keywords: Cnaphalocrocis medinalis; RNA interference; UDP-N-acetylglucosamine pyrophosphorylase; expression pattern; gene cloning.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cloning, Molecular / methods*
  • Gene Expression Regulation, Developmental
  • Gene Expression Regulation, Enzymologic
  • Inactivation, Metabolic
  • Insect Proteins / genetics
  • Insect Proteins / metabolism
  • Moths / enzymology
  • Moths / genetics
  • Moths / growth & development*
  • Nucleotidyltransferases / genetics*
  • Nucleotidyltransferases / metabolism*
  • Open Reading Frames
  • Oryza / parasitology
  • RNA Interference
  • Tissue Distribution

Substances

  • Insect Proteins
  • Nucleotidyltransferases
  • UDPacetylglucosamine pyrophosphorylase