Exploration of RNA-Seq data to identify a potential pathogen of the leaf-mining moth, Stomphastis thraustica (Meyrick, 1908) (Lepidoptera: Gracillariidae)

Abstract

The leaf-mining moth, Stomphastis thraustica (Meyrick, 1908) was imported to Australia as a potential biological control agent of an exotic weed, bellyache bush (Jatropha gossypiifolia), from Peru. The insect colony has been maintained in the quarantine facility for over eight years but recently, significant mortality was observed in the culture. The larvae demonstrated swollen intersegments with a fragile integument. The infected larvae are cloudy muted green or yellowish whereas a healthy late instar larva is a vivid green. They slowly dehydrate and eventually die, at which point the larval body becomes rubbery and turns to black. We used next generation sequencing to identify the cause of mortality in the insects. Total RNA was extracted from 20 larvae in two cohorts, one with and one without apparent symptoms of disease, for deep sequencing on NovaSeq platform after eukaryote ribosomal RNA depletion. We identified several non-insect sequences belonging to viruses, bacteria, and fungi, but none of those showed significant abundance or enrichment in the infected dataset. The sequences related to a unicellular yeast, Saccharomyces cerevisiae, and they were among the highly expressed non-insect contigs; more than 5% of reads in both libraries mapped to the genome of this opportunistic microorganism.