PhytophthoraV1, Main, Exploration, bibRecord, 000117

Molecular assays to detect the presence and viability of Phytophthora ramorum and Grosmannia clavigera.

Identifieur interne : 000117 ( Main/Exploration ); précédent : 000116; suivant : 000118

Molecular assays to detect the presence and viability of Phytophthora ramorum and Grosmannia clavigera.

Auteurs : Barbara Wong [Canada] ; Isabel Leal [Canada] ; Nicolas Feau [Canada] ; Angela Dale [Canada] ; Adnan Uzunovic [Canada] ; Richard C. Hamelin [Canada]

Source :

PloS one [ 1932-6203 ] ; 2020.

RBID : pubmed:32023247

Descripteurs français

KwdFr :
- ADN fongique (analyse), ARN fongique (analyse), Bois (microbiologie), Maladies des plantes (microbiologie), Ophiostomatales (cytologie), Ophiostomatales (génétique), Ophiostomatales (isolement et purification), Phytophthora (cytologie), Phytophthora (génétique), Phytophthora (isolement et purification), Réaction de polymérisation en chaîne (MeSH), Survie cellulaire (MeSH).
MESH :
- analyse : ADN fongique, ARN fongique.
- cytologie : Ophiostomatales, Phytophthora.
- génétique : Ophiostomatales, Phytophthora.
- isolement et purification : Ophiostomatales, Phytophthora.
- microbiologie : Bois, Maladies des plantes.
- Réaction de polymérisation en chaîne, Survie cellulaire.

English descriptors

KwdEn :
- Cell Survival (MeSH), DNA, Fungal (analysis), Ophiostomatales (cytology), Ophiostomatales (genetics), Ophiostomatales (isolation & purification), Phytophthora (cytology), Phytophthora (genetics), Phytophthora (isolation & purification), Plant Diseases (microbiology), Polymerase Chain Reaction (MeSH), RNA, Fungal (analysis), Wood (microbiology).
MESH :
- chemical , analysis : DNA, Fungal, RNA, Fungal.
- cytology : Ophiostomatales, Phytophthora.
- genetics : Ophiostomatales, Phytophthora.
- isolation & purification : Ophiostomatales, Phytophthora.
- microbiology : Plant Diseases, Wood.
- Cell Survival, Polymerase Chain Reaction.

Abstract

Wood and wood products can harbor microorganisms that can raise phytosanitary concerns in countries importing or exporting these products. To evaluate the efficacy of wood treatment on the survival of microorganisms of phytosanitary concern the method of choice is to grow microbes in petri dishes for subsequent identification. However, some plant pathogens are difficult or impossible to grow in axenic cultures. A molecular methodology capable of detecting living fungi and fungus-like organisms in situ can provide a solution. RNA represents the transcription of genes and can become rapidly unstable after cell death, providing a proxy measure of viability. We designed and used RNA-based molecular diagnostic assays targeting genes essential to vital processes and assessed their presence in wood colonized by fungi and oomycetes through reverse transcription and real-time polymerase chain reaction (PCR). A stability analysis was conducted by comparing the ratio of mRNA to gDNA over time following heat treatment of mycelial cultures of the Oomycete Phytophthora ramorum and the fungus Grosmannia clavigera. The real-time PCR results indicated that the DNA remained stable over a period of 10 days post treatment in heat-treated samples, whereas mRNA could not be detected after 24 hours for P. ramorum or 96 hours for G. clavigera. Therefore, this method provides a reliable way to evaluate the viability of these pathogens and offers a potential way to assess the effectiveness of existing and emerging wood treatments. This can have important phytosanitary impacts on assessing both timber and non-timber forest products of commercial value in international wood trade.

DOI: 10.1371/journal.pone.0221742
PubMed: 32023247
PubMed Central: PMC7001964

Affiliations:

Links toward previous steps (curation, corpus...)

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Le document en format XML

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<term>Phytophthora (isolation & purification)</term>
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<term>Maladies des plantes (microbiologie)</term>
<term>Ophiostomatales (cytologie)</term>
<term>Ophiostomatales (génétique)</term>
<term>Ophiostomatales (isolement et purification)</term>
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<term>Phytophthora (génétique)</term>
<term>Phytophthora (isolement et purification)</term>
<term>Réaction de polymérisation en chaîne (MeSH)</term>
<term>Survie cellulaire (MeSH)</term>
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<term>RNA, Fungal</term>
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<term>ARN fongique</term>
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<front><div type="abstract" xml:lang="en">Wood and wood products can harbor microorganisms that can raise phytosanitary concerns in countries importing or exporting these products. To evaluate the efficacy of wood treatment on the survival of microorganisms of phytosanitary concern the method of choice is to grow microbes in petri dishes for subsequent identification. However, some plant pathogens are difficult or impossible to grow in axenic cultures. A molecular methodology capable of detecting living fungi and fungus-like organisms in situ can provide a solution. RNA represents the transcription of genes and can become rapidly unstable after cell death, providing a proxy measure of viability. We designed and used RNA-based molecular diagnostic assays targeting genes essential to vital processes and assessed their presence in wood colonized by fungi and oomycetes through reverse transcription and real-time polymerase chain reaction (PCR). A stability analysis was conducted by comparing the ratio of mRNA to gDNA over time following heat treatment of mycelial cultures of the Oomycete Phytophthora ramorum and the fungus Grosmannia clavigera. The real-time PCR results indicated that the DNA remained stable over a period of 10 days post treatment in heat-treated samples, whereas mRNA could not be detected after 24 hours for P. ramorum or 96 hours for G. clavigera. Therefore, this method provides a reliable way to evaluate the viability of these pathogens and offers a potential way to assess the effectiveness of existing and emerging wood treatments. This can have important phytosanitary impacts on assessing both timber and non-timber forest products of commercial value in international wood trade.</div>
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<Abstract><AbstractText>Wood and wood products can harbor microorganisms that can raise phytosanitary concerns in countries importing or exporting these products. To evaluate the efficacy of wood treatment on the survival of microorganisms of phytosanitary concern the method of choice is to grow microbes in petri dishes for subsequent identification. However, some plant pathogens are difficult or impossible to grow in axenic cultures. A molecular methodology capable of detecting living fungi and fungus-like organisms in situ can provide a solution. RNA represents the transcription of genes and can become rapidly unstable after cell death, providing a proxy measure of viability. We designed and used RNA-based molecular diagnostic assays targeting genes essential to vital processes and assessed their presence in wood colonized by fungi and oomycetes through reverse transcription and real-time polymerase chain reaction (PCR). A stability analysis was conducted by comparing the ratio of mRNA to gDNA over time following heat treatment of mycelial cultures of the Oomycete Phytophthora ramorum and the fungus Grosmannia clavigera. The real-time PCR results indicated that the DNA remained stable over a period of 10 days post treatment in heat-treated samples, whereas mRNA could not be detected after 24 hours for P. ramorum or 96 hours for G. clavigera. Therefore, this method provides a reliable way to evaluate the viability of these pathogens and offers a potential way to assess the effectiveness of existing and emerging wood treatments. This can have important phytosanitary impacts on assessing both timber and non-timber forest products of commercial value in international wood trade.</AbstractText>
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<AffiliationInfo><Affiliation>Department of Forest and Conservation Sciences, University of British Columbia, Vancouver, BC, Canada.</Affiliation>
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Molecular assays to detect the presence and viability of Phytophthora ramorum and Grosmannia clavigera.

Molecular assays to detect the presence and viability of Phytophthora ramorum and Grosmannia clavigera.

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