Preprint Article Version 1 This version is not peer-reviewed

Next-generation sequencing to investigate existing and new insect associations with phytopathogenic fungal propagules

Version 1 : Received: 15 October 2018 / Approved: 15 October 2018 / Online: 15 October 2018 (18:52:27 CEST)

How to cite: Tremblay, É.D.; Kimoto, T.; Bérubé, J.A.; Bilodeau, G.J. Next-generation sequencing to investigate existing and new insect associations with phytopathogenic fungal propagules. Preprints 2018, 2018100333 (doi: 10.20944/preprints201810.0333.v1). Tremblay, É.D.; Kimoto, T.; Bérubé, J.A.; Bilodeau, G.J. Next-generation sequencing to investigate existing and new insect associations with phytopathogenic fungal propagules. Preprints 2018, 2018100333 (doi: 10.20944/preprints201810.0333.v1).

Abstract

Understanding ecological interactions is a key in managing phytopathology. Although entomologists rely mostly on both traditional molecular methods and morphological characteristics to identify pests, next-generation sequencing is becoming the go-to avenue for scientists studying fungal and oomycete phytopathogens. These organisms sometimes infect plants together with insects. There are many relationships yet to be discovered and much to learn about how these organisms interact with one another. Considering the growing number of exotic insect introductions in Canada, a high-throughput strategy for screening those insects is already implemented by the Canadian Food Inspection Agency (CFIA). However, no plan is deployed to investigate the phytopathogenic fungal and oomycete species interacting with insects. Metagenomics analysis was performed on the preservation fluids from CFIA’s insect traps across Canada. Using the Ion Torrent PGM technology and fusion primers for multiplexing and indexing, community profiling was conducted on the different semiochemicals used in the insect traps and the various areas where these traps were placed. Internal transcribed spacer 1 (fungi and oomycetes) and adenosine triphosphate synthase subunit 9-nicotinamide adenine dinucleotide dehydrogenase subunit 9 spacer amplicons were generated. Although direct links between organisms could not be established, moderately phytopathogenic fungi (e.g., Leptographium spp. and Meria laricis) and oomycetes (mainly Peronospora spp. and Pythium spp.) unique to every type of semiochemical were discovered. The entomopathogenic yeast Candida michaelii was also detected. This project demonstrated our ability to screen for unwanted species faster and at a higher scale and throughput than traditional pathogen diagnostic techniques. Additionally, minimal modifications to this approach would allow it to be used in other phytopathology fields.

Subject Areas

Insects, Forest, Fungi, ITS1, metagenomics, NGS, Oomycete, Phytopathogens, Phytophthora, vectors

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