Cloning, expression, and functional analysis of two acetylcholinesterase genes in Spodoptera litura (Lepidoptera: Noctuidae)

Comp Biochem Physiol B Biochem Mol Biol. 2017 Apr:206:16-25. doi: 10.1016/j.cbpb.2017.01.007. Epub 2017 Jan 19.

Abstract

Two acetylcholinesterase genes (SlAce1 and SlAce2) were cloned from Spodoptera litura, which is an important pest that causes widespread economic damage to vegetables and ornamental plants. We analyzed their expression patterns and compared their biological functions by using RNA interference. Our results showed that SlAce1 and SlAce2 cDNA contains 2085bp and 1917bp nucleotides and encoding proteins of 694 and 638 amino acid residues, respectively. Phylogenic analysis indicated that the lineage of SlAce genes and SlAce1 was completely different from SlAce2. Although both genes were expressed in all developmental stages and majorly in the brain. The expression levels of the both genes were suppressed by inserting their related dsRNA in the 6th instar larvae, which led to 47.3% (SlAce1) and 37.9% (SlAce2) mortality. Interestingly, the suppression of the SlAce2 transcripts also led to significant reductions in the fecundity, hatching, and offspring in the parental generation of S. litura. It is concluded that SlAce2 is responsible for the hydrolysis of acetylcholine and also plays role in female breeding, embryo progress, and the development of progeny. Considerable larval mortality was observed after both AChE genes (i.e. Ace1 and Ace2) were silenced in S. litura confirms its insecticidal effectiveness, which provided a molecular basis in biological pest control approach.

Keywords: Acetylcholinesterase; Gene function; Quantitative real-time PCR; RNA interference; Spodoptera litura.

MeSH terms

  • Acetylcholinesterase / chemistry
  • Acetylcholinesterase / genetics*
  • Acetylcholinesterase / metabolism*
  • Animals
  • Cloning, Molecular
  • Female
  • Gene Expression Regulation, Enzymologic*
  • Gene Silencing
  • Larva
  • Peroxisome Proliferator-Activated Receptors / agonists
  • Peroxisome Proliferator-Activated Receptors / metabolism
  • Phylogeny
  • RNA, Double-Stranded / genetics
  • Spodoptera / classification
  • Spodoptera / enzymology*
  • Spodoptera / genetics*

Substances

  • Peroxisome Proliferator-Activated Receptors
  • RNA, Double-Stranded
  • Acetylcholinesterase