Developing a non-destructive metabarcoding protocol for detection of pest insects in bulk trap catches

Abstract Metabarcoding has the potential to revolutionise insect surveillance by providing high-throughput and cost-effective species identification of all specimens within mixed trap catches. Nevertheless, incorporation of metabarcoding into insect diagnostic laboratories will first require the dev...

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Autores principales:	Jana Batovska, Alexander M. Piper, Isabel Valenzuela, John Paul Cunningham, Mark J. Blacket
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/f68bd94e81da4159a3ab5ab43770f5d7
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spelling	oai:doaj.org-article:f68bd94e81da4159a3ab5ab43770f5d72021-12-02T18:02:48ZDeveloping a non-destructive metabarcoding protocol for detection of pest insects in bulk trap catches10.1038/s41598-021-85855-62045-2322https://doaj.org/article/f68bd94e81da4159a3ab5ab43770f5d72021-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-85855-6https://doaj.org/toc/2045-2322Abstract Metabarcoding has the potential to revolutionise insect surveillance by providing high-throughput and cost-effective species identification of all specimens within mixed trap catches. Nevertheless, incorporation of metabarcoding into insect diagnostic laboratories will first require the development and evaluation of protocols that adhere to the specialised regulatory requirements of invasive species surveillance. In this study, we develop a multi-locus non-destructive metabarcoding protocol that allows sensitive detection of agricultural pests, and subsequent confirmation using traditional diagnostic techniques. We validate this protocol for the detection of tomato potato psyllid (Bactericera cockerelli) and Russian wheat aphid (Diuraphis noxia) within mock communities and field survey traps. We find that metabarcoding can reliably detect target insects within mixed community samples, including specimens that morphological identification did not initially detect, but sensitivity appears inversely related to community size and is impacted by primer biases, target loci, and sample indexing strategy. While our multi-locus approach allowed independent validation of target detection, lack of reference sequences for 18S and 12S restricted its usefulness for estimating diversity in field samples. The non-destructive DNA extraction proved invaluable for resolving inconsistencies between morphological and metabarcoding identification results, and post-extraction specimens were suitable for both morphological re-examination and DNA re-extraction for confirmatory barcoding.Jana BatovskaAlexander M. PiperIsabel ValenzuelaJohn Paul CunninghamMark J. BlacketNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-14 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jana Batovska Alexander M. Piper Isabel Valenzuela John Paul Cunningham Mark J. Blacket Developing a non-destructive metabarcoding protocol for detection of pest insects in bulk trap catches
description	Abstract Metabarcoding has the potential to revolutionise insect surveillance by providing high-throughput and cost-effective species identification of all specimens within mixed trap catches. Nevertheless, incorporation of metabarcoding into insect diagnostic laboratories will first require the development and evaluation of protocols that adhere to the specialised regulatory requirements of invasive species surveillance. In this study, we develop a multi-locus non-destructive metabarcoding protocol that allows sensitive detection of agricultural pests, and subsequent confirmation using traditional diagnostic techniques. We validate this protocol for the detection of tomato potato psyllid (Bactericera cockerelli) and Russian wheat aphid (Diuraphis noxia) within mock communities and field survey traps. We find that metabarcoding can reliably detect target insects within mixed community samples, including specimens that morphological identification did not initially detect, but sensitivity appears inversely related to community size and is impacted by primer biases, target loci, and sample indexing strategy. While our multi-locus approach allowed independent validation of target detection, lack of reference sequences for 18S and 12S restricted its usefulness for estimating diversity in field samples. The non-destructive DNA extraction proved invaluable for resolving inconsistencies between morphological and metabarcoding identification results, and post-extraction specimens were suitable for both morphological re-examination and DNA re-extraction for confirmatory barcoding.
format	article
author	Jana Batovska Alexander M. Piper Isabel Valenzuela John Paul Cunningham Mark J. Blacket
author_facet	Jana Batovska Alexander M. Piper Isabel Valenzuela John Paul Cunningham Mark J. Blacket
author_sort	Jana Batovska
title	Developing a non-destructive metabarcoding protocol for detection of pest insects in bulk trap catches
title_short	Developing a non-destructive metabarcoding protocol for detection of pest insects in bulk trap catches
title_full	Developing a non-destructive metabarcoding protocol for detection of pest insects in bulk trap catches
title_fullStr	Developing a non-destructive metabarcoding protocol for detection of pest insects in bulk trap catches
title_full_unstemmed	Developing a non-destructive metabarcoding protocol for detection of pest insects in bulk trap catches
title_sort	developing a non-destructive metabarcoding protocol for detection of pest insects in bulk trap catches
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/f68bd94e81da4159a3ab5ab43770f5d7
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Developing a non-destructive metabarcoding protocol for detection of pest insects in bulk trap catches

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