Abstract

BackgroundInsects use pheromones, chemical signals that underlie all animal behaviors, for communication and for attracting mates. Synthetic pheromones are widely used in pest control strategies because they are environmentally safe. The production of insect pheromones in transgenic plants, which could be more economical and effective in producing isomerically pure compounds, has recently been successfully demonstrated. This research requires information regarding the pheromone biosynthetic pathways and the characterization of pheromone biosynthetic enzymes (PBEs). We used Illumina sequencing to characterize the pheromone gland (PG) transcriptome of the Pyralid moth, Ephestia cautella, a destructive storage pest, to reveal putative candidate genes involved in pheromone biosynthesis, release, transport and degradation.ResultsWe isolated the E. cautella pheromone compound as (Z,E)-9,12-tetradecadienyl acetate, and the major pheromone precursors 16:acyl, 14:acyl, E14-16:acyl, E12-14:acyl and Z9,E12-14:acyl. Based on the abundance of precursors, two possible pheromone biosynthetic pathways are proposed. Both pathways initiate from C16:acyl-CoA, with one involving ∆14 and ∆9 desaturation to generate Z9,E12-14:acyl, and the other involving the chain shortening of C16:acyl-CoA to C14:acyl-CoA, followed by ∆12 and ∆9 desaturation to generate Z9,E12-14:acyl-CoA. Then, a final reduction and acetylation generates Z9,E12-14:OAc. Illumina sequencing yielded 83,792 transcripts, and we obtained a PG transcriptome of ~49.5 Mb. A total of 191 PBE transcripts, which included pheromone biosynthesis activating neuropeptides, fatty acid transport proteins, acetyl-CoA carboxylases, fatty acid synthases, desaturases, β-oxidation enzymes, fatty acyl-CoA reductases (FARs) and fatty acetyltransferases (FATs), were selected from the dataset. A comparison of the E. cautella transcriptome data with three other Lepidoptera PG datasets revealed that 45 % of the sequences were shared. Phylogenetic trees were constructed for desaturases, FARs and FATs, and transcripts that clustered with the ∆14, ∆12 and ∆9 desaturases, PG-specific FARs and potential candidate FATs, respectively, were identified. Transcripts encoding putative pheromone degrading enzymes, and candidate pheromone carrier and receptor proteins expressed in the E. cautella PG, were also identified.ConclusionsOur study provides important background information on the enzymes involved in pheromone biosynthesis. This information will be useful for the in vitro production of E. cautella sex pheromones and may provide potential targets for disrupting the pheromone-based communication system of E. cautella to prevent infestations.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1710-2) contains supplementary material, which is available to authorized users.

Highlights

  • Insects use pheromones, chemical signals that underlie all animal behaviors, for communication and for attracting mates

  • Sex pheromone gland (PG) extraction and fatty-acyl precursor analysis Analysis of the E. cautella PGs excised at the calling period (2-day-old, at mid-scotophase) showed the presence of the compound (Z,12- tetradecadienyl acetate (E)-9),12-tetradecadienyl acetate (Z9,E1214:OAc) by their GC retention times (18.19 min) and mass spectra [ion fragment of m/z 61, a characteristic of acetate compounds (CH3COOH2+) and diagnostic ion at m/z 192] in comparison with those of authentic pheromone samples (Fig. 1)

  • Our study provides comprehensive information on the pheromone molecules, biosynthetic pathways and genes expressed in the PG that are related to pheromone biosynthesis, degradation, transport and release

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Summary

Introduction

Chemical signals that underlie all animal behaviors, for communication and for attracting mates. As the first step in pheromone biosynthesis, carboxylation of acetyl-CoA to malonyl-CoA is catalyzed by ACC [10] This is followed by fatty acid synthase (FAS) activity to produce saturated fatty acids (C18:0 and C16:0) using malonyl-CoA as the substrate. Once specific unsaturated pheromone precursors are formed, the terminal carboxyl group is modified to form one of the functional groups, alcohol, aldehyde or acetate ester (OH, CHO or OAc, respectively), and is catalyzed by fatty acyl reductase (FAR), aldehyde reductase (AR) or fatty acetyltransferase (FAT), respectively [3,4,5, 10]. The molecular characterizations of other critical enzymes in the pheromone biosynthetic pathway, such as ACC, FAS, and several β-oxidation and acetylation enzymes, have not been characterized at the enzymatic level in insects

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