Characterisation of the Small RNAs in the Biomedically Important Green-Bottle Blowfly Lucilia Sericata

PLoS One. 2015 Mar 24;10(3):e0122203. doi: 10.1371/journal.pone.0122203. eCollection 2015.

Abstract

Background: The green bottle fly maggot, Lucilia sericata, is a species with importance in medicine, agriculture and forensics. Improved understanding of this species' biology is of great potential benefit to many research communities. MicroRNAs (miRNA) are a short non-protein coding regulatory RNA, which directly regulate a host of protein coding genes at the translational level. They have been shown to have developmental and tissue specific distributions where they impact directly on gene regulation. In order to improve understanding of the biology of L. sericata maggots we have performed small RNA-sequencing of their secretions and tissue at different developmental stages.

Results: We have successfully isolated RNA from the secretions of L. sericata maggots. Illumina small RNA-sequencing of these secretions and the three tissues (crop, salivary gland, gut) revealed that the most common small RNA fragments were derived from ribosomal RNA and transfer RNAs of both insect and bacterial origins. These RNA fragments were highly specific, with the most common tRNAs, such as GlyGCC, predominantly represented by reads derived from the 5' end of the mature maggot tRNA. Each library also had a unique profile of miRNAs with a high abundance of miR-10-5p in the maggot secretions and gut and miR-8 in the food storage organ the crop and salivary glands. The pattern of small RNAs in the bioactive maggot secretions suggests they originate from a combination of saliva, foregut and hindgut tissues. Droplet digital RT-PCR validation of the RNA-sequencing data shows that not only are there differences in the tissue profiles for miRNAs and small RNA fragments but that these are also modulated through developmental stages of the insect.

Conclusions: We have identified the small-RNAome of the medicinal maggots L. sericata and shown that there are distinct subsets of miRNAs expressed in specific tissues that also alter during the development of the insect. Furthermore there are very specific RNA fragments derived from other non-coding RNAs present in tissues and in the secretions. This new knowledge has applicability in diverse research fields including wound healing, agriculture and forensics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Bodily Secretions / chemistry
  • Diptera / genetics*
  • Gastrointestinal Tract / chemistry
  • Gene Expression Regulation / genetics*
  • Gene Library
  • Larva / chemistry
  • MicroRNAs / genetics*
  • Molecular Sequence Data
  • RNA, Ribosomal / genetics
  • RNA, Transfer / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, RNA

Substances

  • MicroRNAs
  • RNA, Ribosomal
  • RNA, Transfer

Grant support

Funding was received from internal University of Auckland research grants and from the Maurice Wilkins Centre for Molecular Biodiscovery (www.mauricewilkinscentre.org). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.