Development and evaluation of a recombinase polymerase amplification assay for rapid detection of strawberry red stele pathogen
Abstract Phytophthora fragariae causes drastic damage in strawberry crops. P. fragariae infects strawberry roots and causes red stele root rot. Although P. fragariae is a quarantine organism, its spread in Finland continues as more and more fields contract the disease. The spread can be halted throu...
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doaj-ea44f2eb55644da1ba7a6564e236ac5a2020-11-25T03:33:05ZengBMCPhytopathology Research2524-41672020-08-012111210.1186/s42483-020-00069-4Development and evaluation of a recombinase polymerase amplification assay for rapid detection of strawberry red stele pathogenMustafa Ahmad Munawar0Anna Toljamo1Frank Martin2Elina Oksanen3Harri Kokko4Department of Environmental and Biological Sciences, University of Eastern FinlandDepartment of Environmental and Biological Sciences, University of Eastern FinlandUnited States Department of Agriculture, ARSDepartment of Environmental and Biological Sciences, University of Eastern FinlandDepartment of Environmental and Biological Sciences, University of Eastern FinlandAbstract Phytophthora fragariae causes drastic damage in strawberry crops. P. fragariae infects strawberry roots and causes red stele root rot. Although P. fragariae is a quarantine organism, its spread in Finland continues as more and more fields contract the disease. The spread can be halted through developing rapid and reliable detection assays. We have developed a rapid recombinase polymerase amplification (RPA) assay for P. fragariae targeting the Phytophthora mitochondrial DNA intergenic atp9-nad9 marker. The assay is DNA-extraction free and capable of detecting as low as 10 fg of P. fragariae genomic DNA. We found the assay reliable for diagnosing field plants when samples are adequately collected. We also applied the RPA assay to the detection of the pathogen in the soil through coupling the assay with baiting with the host plant. The results suggest that if only a small number of samples are analysed, the baiting results will not be reliable.http://link.springer.com/article/10.1186/s42483-020-00069-4Phytophthora fragariaeRed steleRecombinase polymerase amplificationCrude macerationBaitingIndicator plants |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Mustafa Ahmad Munawar Anna Toljamo Frank Martin Elina Oksanen Harri Kokko |
spellingShingle |
Mustafa Ahmad Munawar Anna Toljamo Frank Martin Elina Oksanen Harri Kokko Development and evaluation of a recombinase polymerase amplification assay for rapid detection of strawberry red stele pathogen Phytopathology Research Phytophthora fragariae Red stele Recombinase polymerase amplification Crude maceration Baiting Indicator plants |
author_facet |
Mustafa Ahmad Munawar Anna Toljamo Frank Martin Elina Oksanen Harri Kokko |
author_sort |
Mustafa Ahmad Munawar |
title |
Development and evaluation of a recombinase polymerase amplification assay for rapid detection of strawberry red stele pathogen |
title_short |
Development and evaluation of a recombinase polymerase amplification assay for rapid detection of strawberry red stele pathogen |
title_full |
Development and evaluation of a recombinase polymerase amplification assay for rapid detection of strawberry red stele pathogen |
title_fullStr |
Development and evaluation of a recombinase polymerase amplification assay for rapid detection of strawberry red stele pathogen |
title_full_unstemmed |
Development and evaluation of a recombinase polymerase amplification assay for rapid detection of strawberry red stele pathogen |
title_sort |
development and evaluation of a recombinase polymerase amplification assay for rapid detection of strawberry red stele pathogen |
publisher |
BMC |
series |
Phytopathology Research |
issn |
2524-4167 |
publishDate |
2020-08-01 |
description |
Abstract Phytophthora fragariae causes drastic damage in strawberry crops. P. fragariae infects strawberry roots and causes red stele root rot. Although P. fragariae is a quarantine organism, its spread in Finland continues as more and more fields contract the disease. The spread can be halted through developing rapid and reliable detection assays. We have developed a rapid recombinase polymerase amplification (RPA) assay for P. fragariae targeting the Phytophthora mitochondrial DNA intergenic atp9-nad9 marker. The assay is DNA-extraction free and capable of detecting as low as 10 fg of P. fragariae genomic DNA. We found the assay reliable for diagnosing field plants when samples are adequately collected. We also applied the RPA assay to the detection of the pathogen in the soil through coupling the assay with baiting with the host plant. The results suggest that if only a small number of samples are analysed, the baiting results will not be reliable. |
topic |
Phytophthora fragariae Red stele Recombinase polymerase amplification Crude maceration Baiting Indicator plants |
url |
http://link.springer.com/article/10.1186/s42483-020-00069-4 |
work_keys_str_mv |
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