Genome-Informed Real-Time Polymerase Chain Reaction for Detection of Erwinia pyrifoliae, the Causal Agent of Asian Pear Blight

Erwinia pyrifoliae causes blight symptoms on pears (Pyrus spp.), apple (Malus spp.), and strawberry (Fragaria × ananassa), with up to 40% losses in strawberry observed during a 2013 outbreak in the Netherlands. These commodities are of economic importance in the United States. Introduction and estab...

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Main Authors: Jarred Yasuhara-Bell, Vessela Mavrodieva, Yazmín Rivera
Format: Article
Language:English
Published: The American Phytopathological Society 2025-06-01
Series:PhytoFrontiers
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Online Access:https://apsjournals.apsnet.org/doi/10.1094/PHYTOFR-08-24-0091-FI
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author Jarred Yasuhara-Bell
Vessela Mavrodieva
Yazmín Rivera
author_facet Jarred Yasuhara-Bell
Vessela Mavrodieva
Yazmín Rivera
author_sort Jarred Yasuhara-Bell
collection DOAJ
description Erwinia pyrifoliae causes blight symptoms on pears (Pyrus spp.), apple (Malus spp.), and strawberry (Fragaria × ananassa), with up to 40% losses in strawberry observed during a 2013 outbreak in the Netherlands. These commodities are of economic importance in the United States. Introduction and establishment of E. pyrifoliae could result in huge economic losses, thus creating a need for appropriate surveillance and disease management. The availability of validated diagnostic tools is the cornerstone of successful surveillance, quarantine, and eradication measures. In this study, assays based on two different E. pyrifoliae-specific targets were validated thoroughly. The hrpW target was described previously, whereas the hsvC target was established in this study. Both were duplexed with either a 16S rDNA (bacteria) or 18S rDNA (plant) internal control to accommodate different sample types and extractions. The hrpW and hsvC targets showed similar performance, with a limit of detection of approximately five genome equivalent copies per reaction with 100% positivity, demonstrating high sensitivity. The assays also demonstrated high analytical and diagnostic specificity. Positive results were obtained for eight known E. pyrifoliae isolates, comprising Korean and Japanese isolates, and no cross-reactions were observed with other Erwinia spp. tested, except closely related E. uzenensis, as well as 74 other pathogens and two commercial biocontrol agents. Symptomatic strawberries were found in the United States, and these assays were able to confirm the first report of E. pyrifoliae. These assays were deployed successfully and are beneficial tools that support efforts to protect U.S. agriculture and facilitate safe trade. [Figure: see text] Copyright © 2025 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.
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spelling doaj-art-1127b672f0df44c9ba5c29aaee14c7842025-08-20T02:38:01ZengThe American Phytopathological SocietyPhytoFrontiers2690-54422025-06-015217418610.1094/PHYTOFR-08-24-0091-FIGenome-Informed Real-Time Polymerase Chain Reaction for Detection of Erwinia pyrifoliae, the Causal Agent of Asian Pear BlightJarred Yasuhara-Bell0Vessela Mavrodieva1Yazmín Rivera2Plant Pathogen Confirmatory Diagnostics Laboratory (PPCDL), Science and Technology (S&T), Plant Protection and Quarantine (PPQ), Animal and Plant Health Inspection Service (APHIS), U.S. Department of Agriculture (USDA), Laurel, MD 20708, U.S.A.Plant Pathogen Confirmatory Diagnostics Laboratory (PPCDL), Science and Technology (S&T), Plant Protection and Quarantine (PPQ), Animal and Plant Health Inspection Service (APHIS), U.S. Department of Agriculture (USDA), Laurel, MD 20708, U.S.A.Plant Pathogen Confirmatory Diagnostics Laboratory (PPCDL), Science and Technology (S&T), Plant Protection and Quarantine (PPQ), Animal and Plant Health Inspection Service (APHIS), U.S. Department of Agriculture (USDA), Laurel, MD 20708, U.S.A.Erwinia pyrifoliae causes blight symptoms on pears (Pyrus spp.), apple (Malus spp.), and strawberry (Fragaria × ananassa), with up to 40% losses in strawberry observed during a 2013 outbreak in the Netherlands. These commodities are of economic importance in the United States. Introduction and establishment of E. pyrifoliae could result in huge economic losses, thus creating a need for appropriate surveillance and disease management. The availability of validated diagnostic tools is the cornerstone of successful surveillance, quarantine, and eradication measures. In this study, assays based on two different E. pyrifoliae-specific targets were validated thoroughly. The hrpW target was described previously, whereas the hsvC target was established in this study. Both were duplexed with either a 16S rDNA (bacteria) or 18S rDNA (plant) internal control to accommodate different sample types and extractions. The hrpW and hsvC targets showed similar performance, with a limit of detection of approximately five genome equivalent copies per reaction with 100% positivity, demonstrating high sensitivity. The assays also demonstrated high analytical and diagnostic specificity. Positive results were obtained for eight known E. pyrifoliae isolates, comprising Korean and Japanese isolates, and no cross-reactions were observed with other Erwinia spp. tested, except closely related E. uzenensis, as well as 74 other pathogens and two commercial biocontrol agents. Symptomatic strawberries were found in the United States, and these assays were able to confirm the first report of E. pyrifoliae. These assays were deployed successfully and are beneficial tools that support efforts to protect U.S. agriculture and facilitate safe trade. [Figure: see text] Copyright © 2025 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.https://apsjournals.apsnet.org/doi/10.1094/PHYTOFR-08-24-0091-FIAsian pear blightbacterial shoot blight of peardetectionErwinia pyrifoliaePCR
spellingShingle Jarred Yasuhara-Bell
Vessela Mavrodieva
Yazmín Rivera
Genome-Informed Real-Time Polymerase Chain Reaction for Detection of Erwinia pyrifoliae, the Causal Agent of Asian Pear Blight
PhytoFrontiers
Asian pear blight
bacterial shoot blight of pear
detection
Erwinia pyrifoliae
PCR
title Genome-Informed Real-Time Polymerase Chain Reaction for Detection of Erwinia pyrifoliae, the Causal Agent of Asian Pear Blight
title_full Genome-Informed Real-Time Polymerase Chain Reaction for Detection of Erwinia pyrifoliae, the Causal Agent of Asian Pear Blight
title_fullStr Genome-Informed Real-Time Polymerase Chain Reaction for Detection of Erwinia pyrifoliae, the Causal Agent of Asian Pear Blight
title_full_unstemmed Genome-Informed Real-Time Polymerase Chain Reaction for Detection of Erwinia pyrifoliae, the Causal Agent of Asian Pear Blight
title_short Genome-Informed Real-Time Polymerase Chain Reaction for Detection of Erwinia pyrifoliae, the Causal Agent of Asian Pear Blight
title_sort genome informed real time polymerase chain reaction for detection of erwinia pyrifoliae the causal agent of asian pear blight
topic Asian pear blight
bacterial shoot blight of pear
detection
Erwinia pyrifoliae
PCR
url https://apsjournals.apsnet.org/doi/10.1094/PHYTOFR-08-24-0091-FI
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