Development associated profiling of chitinase and microRNA of Helicoverpa armigera identified chitinase repressive microRNA

Sci Rep. 2013:3:2292. doi: 10.1038/srep02292.

Abstract

Expression of chitinase is developmentally regulated in insects in consonance with their molting process. During the larval-larval metamorphosis in Helicoverpa armigera, chitinase gene expression varies from high to negligible. In the five-day metamorphic course of fifth-instar larvae, chitinase transcript is least abundant on third day and maximal on fifth day. MicroRNA library prepared from these highest and lowest chitinase-expressing larval stages resulted in isolation of several miRNAs. In silico analysis of sequenced miRNAs revealed three miRNAs having sequence similarity to 3'UTR of chitinase. Gene-targeted specific action of these miRNAs, was investigated by luciferase reporter having 3'UTR of chitinase. Only one of three miRNAs, miR-24, inhibited luciferase expression. Further, a day-wise in vivo quantification of miR-24 in fifth-instar larvae revealed a negative correlation with corresponding chitinase transcript abundance. The force-feeding of synthetic miR-24 induced significant morphological aberrations accompanied with arrest of molting. These miR-24 force-fed larvae revealed significantly reduced chitinase transcript abundance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions
  • Animals
  • Base Sequence
  • Cell Line
  • Chitinases / chemistry
  • Chitinases / genetics*
  • Chitinases / metabolism
  • Gene Expression Profiling*
  • Gene Expression Regulation, Developmental
  • Larva / genetics
  • Larva / metabolism
  • Metamorphosis, Biological / genetics
  • MicroRNAs / genetics
  • Molecular Sequence Data
  • Moths / enzymology
  • Moths / genetics*
  • Phenotype
  • Reproducibility of Results
  • Time Factors
  • Transcription, Genetic

Substances

  • 3' Untranslated Regions
  • MicroRNAs
  • Chitinases