Unveiling invasive insect threats to plant biodiversity: Leveraging eDNA metabarcoding and saturated salt trap solutions for biosurveillance

18. Hakimzadeh A, Asbun AA, Albanese D, Bernard M, Buchner D, Callahan B, et al. A pile of pipelines: an overview of the bioinformatics software for metabarcoding data analyses. Preprints; 2023 Feb. https:// doi.org/10.22541/au.167610608.87815963/v1 19. Porter TM, Hajibabaei M. MetaWorks: A flexible, scalable bioinformatic pipeline for high-throughput multi-marker biodiversity assessments. PLOS ONE. 2022; 17: e0274260. https://doi.org/10.1371/ journal.pone.0274260 PMID: 36174014 20. Ratnasingham S. mBRAVE: The Multiplex Barcode Research And Visualization Environment. Biodiver- sity Information Science and Standards. 2019; 3: e37986. https://doi.org/10.3897/biss.3.37986 21. mBRAVE—Metabarcoding at Scale. [cited 24 Feb 2023]. Available: https://www.mbrave.net/ 22. Wang Q, Garrity GM, Tiedje JM, Cole JR. Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol. 2007; 73: 5261–5267. https://doi. org/10.1128/AEM.00062-07 PMID: 17586664 23. Young RG, Milia´n-Garcı´a Y, Yu J, Bullas-Appleton E, Hanner RH. Biosurveillance for invasive insect pest species using an environmental DNA metabarcoding approach and a high salt trap collection fluid. Ecology and Evolution. 2020. https://doi.org/10.1002/ece3.7113 PMID: 33613989 24. Milia´ n-Garcı´a Y. Modified Qiagen DNeasy Blood and Tissue extraction method (Cat. No. / ID: 69506) for eDNA extraction from filters (Nitrocellulose Mixed Ester membrane filters or similar). In: protocols.io [Internet]. 2022 [cited 11 Aug 2022]. Available: https://www.protocols.io/private/

8C4AD99B18E311ED837E0A58A9FEAC02 25. Milia´ n-Garcı´a Y, Janke LAA, Young RG, Ambagala A, Hanner RH. Validation of an Effective Protocol for Culicoides Latreille (Diptera: Ceratopogonidae) Detection Using eDNA Metabarcoding. Insects. 2021; 12: 401. https://doi.org/10.3390/insects12050401 PMID: 33946322 26. 16S Metagenomic Sequencing Library Preparation. In: Illumina [Internet]. [cited 29 Nov 2020]. Avail- able: https://support.illumina.com/downloads/16s_metagenomic_sequencing_library_preparation.html 27. Ewels P, Magnusson M, Lundin S, Ka¨ ller M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics. 2016; 32: 3047–3048. https://doi.org/10.1093/bioinformatics/ btw354 PMID: 27312411 28. Ratnasingham S, Hebert PDN. A DNA-Based Registry for All Animal Species: The Barcode Index Num- ber (BIN) System. PLOS ONE. 2013; 8: e66213. https://doi.org/10.1371/journal.pone.0066213 PMID: 23861743 29. Chao A, Chiu C-H, Jost L. Unifying Species Diversity, Phylogenetic Diversity, Functional Diversity, and Related Similarity and Differentiation Measures Through Hill Numbers. Annual Review of Ecology, Evo- lution, and Systematics. 2014; 45: 297–324. https://doi.org/10.1146/annurev-ecolsys-120213-091540 30. Bullas-Appleton E, Kimoto T, Turgeon JJ. Discovery of Trichoferus campestris (Coleoptera: Cerambyci- dae) in Ontario, Canada and first host record in North America. The Canadian Entomologist. 2014; 146: 111–116. https://doi.org/10.4039/tce.2013.61

31. Ministry of Northern Development, Mines, Natural Resources and Foresty. In: https://www.ontario.ca/ page/emerald-ash-borer [Internet]. 20 May 2021 [cited 28 Jun 2022]. Available: http://www.ontario.ca/ page/emerald-ash-borer 32. Haack RA, Jendak E, Houping L, Marchant KR, Petrice TR, Poland TM, et al. The emerald ash borer: a new exotic pest in North America. 2002 [cited 28 Jun 2022]. Available: https://www.nrs.fs.fed.us/pubs/ 1858 33. CABI. Invasive Species Compendium. In: Agrilus planipennis (emerald ash borer) [Internet]. 12 Sep 2020 [cited 28 Jun 2022]. Available: https://www.cabi.org/dmpp/abstract/20063140407 34. Wagner DL. Emerald Ash Borer Threatens Ash-feeding Lepidoptera. News of the Lepidopterists’ Soci- ety. 200749: 3. 35. Hausman CE, Rocha OJ, Jaeger JF. How emerald ash borer facilitates a secondary spread of invasive plant species: impacts of emerald ash borer eradication and tree mortality. Proceedings of the emerald ash borer and Asian longhorned beetle research and technology development meeting, Pittsburgh, Pennsylvania. 2007. pp. 14–15. 36. Hope E, Sun L, McKenney D, Bogdanski B, Pedlar J, MacDonald H, et al. Emerald Ash Borer, Agrilus planipennis: An Economic Analysis of Regulations in Canada. 2020; 40. 37. Hope ES, McKenney DW, Pedlar JH, Lawrence K, MacDonald H. Canadian efforts to slow the spread of Emerald Ash Borer (Agrilus planipennis Fairmaire) are economically efficient. Ecological Economics. 2021; 188: 107126. https://doi.org/10.1016/j.ecolecon.2021.107126

38. Powell KS, Cooper PD, Forneck A. Chapter Four—The Biology, Physiology and Host–Plant Interac- tions of Grape Phylloxera Daktulosphaira vitifoliae. In: Johnson SN, Hiltpold I, Turlings TCJ, editors. Advances in Insect Physiology. Academic Press; 2013. pp. 159–218. https://doi.org/10.1016/B978-0- 12-417165-7.00004–0 PLOS ONE Unveiling invasive insect threats to plant biodiversity PLOS ONE | https://doi.org/10.1371/journal.pone.0290036 August 11, 2023 22 / 24