A Molecular Method to Assess Viability of Phytophthora in Infected Wood Following Phytosanitary Heat Treatment
International trade in wood products is an important component of the global economy. However, wood and wood products may have pests associated with them that could be introduced into importing countries, posing phytosanitary risks and leading to the implementation of regulatory restrictions that af...
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The American Phytopathological Society
2024-12-01
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| Series: | PhytoFrontiers |
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| Online Access: | https://apsjournals.apsnet.org/doi/10.1094/PHYTOFR-05-24-0056-R |
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| author | Isabel Leal Nicolas Feau Adnan Uzunovic Brett Foord Richard C. Hamelin |
| author_facet | Isabel Leal Nicolas Feau Adnan Uzunovic Brett Foord Richard C. Hamelin |
| author_sort | Isabel Leal |
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| description | International trade in wood products is an important component of the global economy. However, wood and wood products may have pests associated with them that could be introduced into importing countries, posing phytosanitary risks and leading to the implementation of regulatory restrictions that affect wood trade. The application of heat to kill wood-associated pests has been a successful phytosanitary method to reduce their spread. To evaluate the efficacy of wood heat treatment to kill fungal and fungus-like pathogens, the method of choice has been to grow organisms in cultures for subsequent identification. However, some plant pathogens can be difficult or impossible to grow in axenic cultures, and a molecular method can still be useful for assessing pathogen viability after heat treatment. RNA is a single-stranded molecule that is responsible for the transcription of genes. Since it becomes rapidly unstable after cell death, it provides a measure of viability. We therefore designed and tested RNA-based molecular diagnostic assays targeting essential genes and assessed their presence after heat treatment in wood colonized by four Phytophthora species of phytosanitary concern (P. ×multiformis, P. cinnamomi, P. lateralis, and P. ramorum) through reverse transcription and real-time polymerase chain reaction (RT-qPCR). Our assays differentiate between genomic and mRNA as the TaqMan probes span exon–intron junctions. We validated these RT-qPCR assays to assess heat treatment efficacy of Phytophthora-inoculated wood. These assays can be very useful tools to assess the effectiveness of current and emerging phytosanitary wood treatments. [Figure: see text] Copyright © 2024 His Majesty the King in Right of Canada, as represented by the Minister of Natural Resources Canada. This is an open access article distributed under the CC BY-NC-ND 4.0 International license. |
| format | Article |
| id | doaj-art-3d7a88962ce14523bb9f17bb5f033dbb |
| institution | DOAJ |
| issn | 2690-5442 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | The American Phytopathological Society |
| record_format | Article |
| series | PhytoFrontiers |
| spelling | doaj-art-3d7a88962ce14523bb9f17bb5f033dbb2025-08-20T02:39:15ZengThe American Phytopathological SocietyPhytoFrontiers2690-54422024-12-014469069910.1094/PHYTOFR-05-24-0056-RA Molecular Method to Assess Viability of Phytophthora in Infected Wood Following Phytosanitary Heat TreatmentIsabel Leal0Nicolas Feau1Adnan Uzunovic2Brett Foord3Richard C. Hamelin4Pacific Forestry Centre, Natural Resources Canada, Victoria, British Columbia, CanadaPacific Forestry Centre, Natural Resources Canada, Victoria, British Columbia, CanadaCanada Wood Group, Vancouver, British Columbia, CanadaPacific Forestry Centre, Natural Resources Canada, Victoria, British Columbia, Canada, RetiredDepartment of Forest and Conservation Sciences, Faculty of Forestry, University of British Columbia, Vancouver, British Columbia, CanadaInternational trade in wood products is an important component of the global economy. However, wood and wood products may have pests associated with them that could be introduced into importing countries, posing phytosanitary risks and leading to the implementation of regulatory restrictions that affect wood trade. The application of heat to kill wood-associated pests has been a successful phytosanitary method to reduce their spread. To evaluate the efficacy of wood heat treatment to kill fungal and fungus-like pathogens, the method of choice has been to grow organisms in cultures for subsequent identification. However, some plant pathogens can be difficult or impossible to grow in axenic cultures, and a molecular method can still be useful for assessing pathogen viability after heat treatment. RNA is a single-stranded molecule that is responsible for the transcription of genes. Since it becomes rapidly unstable after cell death, it provides a measure of viability. We therefore designed and tested RNA-based molecular diagnostic assays targeting essential genes and assessed their presence after heat treatment in wood colonized by four Phytophthora species of phytosanitary concern (P. ×multiformis, P. cinnamomi, P. lateralis, and P. ramorum) through reverse transcription and real-time polymerase chain reaction (RT-qPCR). Our assays differentiate between genomic and mRNA as the TaqMan probes span exon–intron junctions. We validated these RT-qPCR assays to assess heat treatment efficacy of Phytophthora-inoculated wood. These assays can be very useful tools to assess the effectiveness of current and emerging phytosanitary wood treatments. [Figure: see text] Copyright © 2024 His Majesty the King in Right of Canada, as represented by the Minister of Natural Resources Canada. 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-05-24-0056-Rmessenger RNAPhytophphtoraPhytosanitary heat-treatmentRT-qPCRwood products |
| spellingShingle | Isabel Leal Nicolas Feau Adnan Uzunovic Brett Foord Richard C. Hamelin A Molecular Method to Assess Viability of Phytophthora in Infected Wood Following Phytosanitary Heat Treatment PhytoFrontiers messenger RNA Phytophphtora Phytosanitary heat-treatment RT-qPCR wood products |
| title | A Molecular Method to Assess Viability of Phytophthora in Infected Wood Following Phytosanitary Heat Treatment |
| title_full | A Molecular Method to Assess Viability of Phytophthora in Infected Wood Following Phytosanitary Heat Treatment |
| title_fullStr | A Molecular Method to Assess Viability of Phytophthora in Infected Wood Following Phytosanitary Heat Treatment |
| title_full_unstemmed | A Molecular Method to Assess Viability of Phytophthora in Infected Wood Following Phytosanitary Heat Treatment |
| title_short | A Molecular Method to Assess Viability of Phytophthora in Infected Wood Following Phytosanitary Heat Treatment |
| title_sort | molecular method to assess viability of phytophthora in infected wood following phytosanitary heat treatment |
| topic | messenger RNA Phytophphtora Phytosanitary heat-treatment RT-qPCR wood products |
| url | https://apsjournals.apsnet.org/doi/10.1094/PHYTOFR-05-24-0056-R |
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