Characterization and analysis of the transcriptome in Opisina arenosella from different developmental stages using single-molecule real-time transcript sequencing and RNA-seq

Int J Biol Macromol. 2021 Feb 1:169:216-227. doi: 10.1016/j.ijbiomac.2020.12.098. Epub 2020 Dec 16.

Abstract

Opisina arenosella is one of the main pests harming coconut trees. To date, there have been few studies on the molecular genetics, biochemistry and physiology of O. arenosella at the transcriptional level, and there are no available reference genomes. Here, Illumina RNA sequencing combined with PacBio single-molecule real-time analysis was applied to study the transcriptome of this pest at different developmental stages, providing reference data for transcript expression analysis. Twelve samples of O. arenosella from different stages of development were sequenced using Illumina RNA sequencing, and the pooled RNA samples were sequenced with PacBio technology (Iso-Seq). A full-length transcriptome with 41,938 transcripts was captured, and the N50 and N90 lengths were 3543 bp and 1646 bp, respectively. A total of 36,925 transcripts were annotated in public databases, 6493 of which were long noncoding RNAs, while 2510 represented alternative splicing events. There were significant differences in the gene expression profiles at different developmental stages, with high levels of differential gene expression associated with growth, development, carbohydrate metabolism and immunity. This work provides resources and information for the study of the transcriptome and gene function of O. arenosella and provides a valuable foundation for understanding the changes in gene expression during development.

Keywords: Illumina RNA sequencing; Opisina arenosella; Single-molecule real-time sequencing.

MeSH terms

  • Animals
  • Gene Expression / genetics
  • Gene Expression Profiling / methods
  • Lepidoptera / genetics
  • Molecular Sequence Annotation / methods
  • Moths / genetics*
  • RNA / genetics
  • RNA, Long Noncoding / genetics
  • RNA-Seq / methods
  • Sequence Analysis, RNA / methods*
  • Single Molecule Imaging / methods
  • Transcriptome / genetics*

Substances

  • RNA, Long Noncoding
  • RNA