Detection of blueberry stunt phytoplasma in Eastern Canada using cpn60-based molecular diagnostic assays

Abstract Blueberry stunt phytoplasma (BBSP; ‘Candidatus Phytoplasma asteris’) is an insect-vectored plant pathogen that causes severe yield losses in blueberry (Vaccinium corymbosum), which is the most valuable fruit crop in Canada. Rapid, field-based diagnostic assays are desirable tools for the co...

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Autores principales:	Christine Hammond, Edel Pérez-López, Jennifer Town, Charles Vincent, Debra Moreau, Tim Dumonceaux
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4ac88e5e3da3448baaebb47a99101c7e
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spelling	oai:doaj.org-article:4ac88e5e3da3448baaebb47a99101c7e2021-11-14T12:24:21ZDetection of blueberry stunt phytoplasma in Eastern Canada using cpn60-based molecular diagnostic assays10.1038/s41598-021-01439-42045-2322https://doaj.org/article/4ac88e5e3da3448baaebb47a99101c7e2021-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-01439-4https://doaj.org/toc/2045-2322Abstract Blueberry stunt phytoplasma (BBSP; ‘Candidatus Phytoplasma asteris’) is an insect-vectored plant pathogen that causes severe yield losses in blueberry (Vaccinium corymbosum), which is the most valuable fruit crop in Canada. Rapid, field-based diagnostic assays are desirable tools for the control of BBSP, as part of an integrated, proactive approach to production management termed biovigilance. We designed and validated a chaperonin-60 (cpn60)-targeted LAMP assay for detection of BBSP, providing a rapid, low cost, field-deployable diagnostic option. Our validation demonstrates that the assay is reproducible, with high analytical specificity and improved sensitivity when compared with 16S rRNA nested PCR. We applied the validated LAMP assay to nearly 2000 blueberry samples from Québec and Nova Scotia over three growing seasons (2016–2018). Our surveys revealed that BBSP is present in most sites across both provinces, though detection of the pathogen in individual plants varied in different tissues across sampling dates and across years, and evidence of spread between plants was limited. To quantify pathogen load in select plants, we designed additional qPCR and ddPCR assays, also based on cpn60. We found that pathogen load fluctuates in individual plants, both within and between growing seasons. Finally, we designed an interactive map to visualize the results of our surveys. These results provide a validated diagnostic assay that can be used as part of a biovigilance strategy for detecting and controlling infections caused by BBSP.Christine HammondEdel Pérez-LópezJennifer TownCharles VincentDebra MoreauTim DumonceauxNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Christine Hammond Edel Pérez-López Jennifer Town Charles Vincent Debra Moreau Tim Dumonceaux Detection of blueberry stunt phytoplasma in Eastern Canada using cpn60-based molecular diagnostic assays
description	Abstract Blueberry stunt phytoplasma (BBSP; ‘Candidatus Phytoplasma asteris’) is an insect-vectored plant pathogen that causes severe yield losses in blueberry (Vaccinium corymbosum), which is the most valuable fruit crop in Canada. Rapid, field-based diagnostic assays are desirable tools for the control of BBSP, as part of an integrated, proactive approach to production management termed biovigilance. We designed and validated a chaperonin-60 (cpn60)-targeted LAMP assay for detection of BBSP, providing a rapid, low cost, field-deployable diagnostic option. Our validation demonstrates that the assay is reproducible, with high analytical specificity and improved sensitivity when compared with 16S rRNA nested PCR. We applied the validated LAMP assay to nearly 2000 blueberry samples from Québec and Nova Scotia over three growing seasons (2016–2018). Our surveys revealed that BBSP is present in most sites across both provinces, though detection of the pathogen in individual plants varied in different tissues across sampling dates and across years, and evidence of spread between plants was limited. To quantify pathogen load in select plants, we designed additional qPCR and ddPCR assays, also based on cpn60. We found that pathogen load fluctuates in individual plants, both within and between growing seasons. Finally, we designed an interactive map to visualize the results of our surveys. These results provide a validated diagnostic assay that can be used as part of a biovigilance strategy for detecting and controlling infections caused by BBSP.
format	article
author	Christine Hammond Edel Pérez-López Jennifer Town Charles Vincent Debra Moreau Tim Dumonceaux
author_facet	Christine Hammond Edel Pérez-López Jennifer Town Charles Vincent Debra Moreau Tim Dumonceaux
author_sort	Christine Hammond
title	Detection of blueberry stunt phytoplasma in Eastern Canada using cpn60-based molecular diagnostic assays
title_short	Detection of blueberry stunt phytoplasma in Eastern Canada using cpn60-based molecular diagnostic assays
title_full	Detection of blueberry stunt phytoplasma in Eastern Canada using cpn60-based molecular diagnostic assays
title_fullStr	Detection of blueberry stunt phytoplasma in Eastern Canada using cpn60-based molecular diagnostic assays
title_full_unstemmed	Detection of blueberry stunt phytoplasma in Eastern Canada using cpn60-based molecular diagnostic assays
title_sort	detection of blueberry stunt phytoplasma in eastern canada using cpn60-based molecular diagnostic assays
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/4ac88e5e3da3448baaebb47a99101c7e
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Detection of blueberry stunt phytoplasma in Eastern Canada using cpn60-based molecular diagnostic assays

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