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High-Throughput Sequencing to Investigate Phytopathogenic Fungal Propagules Caught in Baited Insect Traps

1
Canadian Food Inspection Agency, 3851 Fallowfield Road, Nepean, ON K2H 8P9, Canada
2
Canadian Food Inspection Agency, 4321 Still Creek Dr, Burnaby, BC V5C 6S7, Canada
3
Natural Resources Canada, Laurentian Forestry Centre, 1055 Du P.E.P.S. Street, P.O. Box 10380 Québec, QC G1V 4C7, Canada
*
Author to whom correspondence should be addressed.
J. Fungi 2019, 5(1), 15; https://doi.org/10.3390/jof5010015
Received: 14 January 2019 / Revised: 2 February 2019 / Accepted: 4 February 2019 / Published: 12 February 2019
(This article belongs to the Special Issue Fungal-Insect Interactions)
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Abstract

Studying the means of dispersal of plant pathogens is crucial to better understand the dynamic interactions involved in plant infections. On one hand, entomologists rely mostly on both traditional molecular methods and morphological characteristics, to identify pests. On the other hand, high-throughput sequencing (HTS) is becoming the go-to avenue for scientists studying phytopathogens. These organisms sometimes infect plants, together with insects. Considering the growing number of exotic insect introductions in Canada, forest pest-management efforts would benefit from the development of a high-throughput strategy to investigate the phytopathogenic fungal and oomycete species interacting with wood-boring insects. We recycled formerly discarded preservative fluids from the Canadian Food Inspection Agency annual survey using insect traps and analysed more than one hundred samples originating from across Canada. Using the Ion Torrent Personal Genome Machine (PGM) HTS technology and fusion primers, we performed metabarcoding to screen unwanted fungi and oomycetes species, including Phytophthora spp. Community profiling was conducted on the four different wood-boring, insect-attracting semiochemicals; although the preservative (contained ethanol) also attracted other insects. Phytopathogenic fungi (e.g., Leptographium spp. and Meria laricis in the pine sawyer semiochemical) and oomycetes (mainly Peronospora spp. and Pythium aff. hypogynum in the General Longhorn semiochemical), solely associated with one of the four types of semiochemicals, were detected. This project demonstrated that the insect traps’ semiochemical microbiome represents a new and powerful matrix for screening phytopathogens. Compared to traditional diagnostic techniques, the fluids allowed for a faster and higher throughput assessment of the biodiversity contained within. Additionally, minimal modifications to this approach would allow it to be used in other phytopathology fields. View Full-Text
Keywords: insects; vectors; forest; fungi; metagenomics; HTS; oomycete insects; vectors; forest; fungi; metagenomics; HTS; oomycete
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Tremblay, É.D.; Kimoto, T.; Bérubé, J.A.; Bilodeau, G.J. High-Throughput Sequencing to Investigate Phytopathogenic Fungal Propagules Caught in Baited Insect Traps. J. Fungi 2019, 5, 15.

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