Development of a high throughput optical density assay to determine fungicide sensitivity of oomycetes.
Identifieur interne : 000799 ( Main/Exploration ); précédent : 000798; suivant : 000800Development of a high throughput optical density assay to determine fungicide sensitivity of oomycetes.
Auteurs : Shannon Hunter [Nouvelle-Zélande] ; Rebecca Mcdougal [Nouvelle-Zélande] ; Mike J. Clearwater [Nouvelle-Zélande] ; Nari Williams [Nouvelle-Zélande] ; Peter Scott [Nouvelle-Zélande]Source :
- Journal of microbiological methods [ 1872-8359 ] ; 2018.
Descripteurs français
- KwdFr :
- Fongicides industriels (pharmacologie), Forêts (MeSH), Maladies des plantes (MeSH), Mycelium (croissance et développement), Nouvelle-Zélande (MeSH), Oomycetes (croissance et développement), Oomycetes (effets des médicaments et des substances chimiques), Phosphites (pharmacologie), Phytophthora (croissance et développement), Phytophthora (effets des médicaments et des substances chimiques), Racines de plante (microbiologie), Tests de sensibilité microbienne (méthodes), Tolérance aux médicaments (MeSH).
- MESH :
- croissance et développement : Mycelium, Oomycetes, Phytophthora.
- effets des médicaments et des substances chimiques : Oomycetes, Phytophthora.
- microbiologie : Racines de plante.
- méthodes : Tests de sensibilité microbienne.
- pharmacologie : Fongicides industriels, Phosphites.
- Forêts, Maladies des plantes, Nouvelle-Zélande, Tolérance aux médicaments.
- Wicri :
- geographic : Nouvelle-Zélande.
English descriptors
- KwdEn :
- Drug Tolerance (MeSH), Forests (MeSH), Fungicides, Industrial (pharmacology), Microbial Sensitivity Tests (methods), Mycelium (growth & development), New Zealand (MeSH), Oomycetes (drug effects), Oomycetes (growth & development), Phosphites (pharmacology), Phytophthora (drug effects), Phytophthora (growth & development), Plant Diseases (MeSH), Plant Roots (microbiology).
- MESH :
- chemical , pharmacology : Fungicides, Industrial, Phosphites.
- geographic : New Zealand.
- drug effects : Oomycetes, Phytophthora.
- growth & development : Mycelium, Oomycetes, Phytophthora.
- methods : Microbial Sensitivity Tests.
- microbiology : Plant Roots.
- Drug Tolerance, Forests, Plant Diseases.
Abstract
A high-throughput assay was developed to screen Phytophthora species for fungicide sensitivity using optical density measurements for unbiased, automated measurement of mycelial growth. The efficacy of the optical density assay (OD) to measure phosphite sensitivity in Phytophthora species was compared to two widely used methods, radial growth (RG) and dry weight (DW) assays. Three isolates of each of Phytophthora cinnamomi, P. multivora and P. pluvialis, with known phosphite exposure and three isolates of each species with no prior phosphite exposure, were screened for phosphite sensitivity using the three assays. Mycelial growth measurements were taken after culturing for 6, 14 and 15 days for the OD, DW and RG assays respectively. Mycelial growth inhibition at 15, 80, 200 and 500 μg/mL phosphite relative to growth on control media was used to determine effective concentration values for 50% growth reduction (EC50). The species varied in their tolerance to phosphite with P. cinnamomi being the least sensitive followed by P. multivora and P. pluvialis. No significant differences in tolerance were found between isolates within the same species using any method. The OD assay produced comparable EC50 values to the RG and DW assays. The growth of the three species was more sensitive to phosphite in the DW than the RG and OD assays, however limited sample throughput and greater variation in measuring small amounts of mycelia in the dry weight assessment increase variability and limits throughput. The OD assay offers a fast method to enable an inventory of chemical resistance and is particularly advantageous for slow growing species as it requires less time and offers greater throughput than existing RG and DW methods.
DOI: 10.1016/j.mimet.2018.10.006
PubMed: 30308213
Affiliations:
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Clearwater</name><affiliation wicri:level="1"><nlm:affiliation>Department of Biology, School of Science, University of Waikato, Hamilton 3216, New Zealand.</nlm:affiliation><country xml:lang="fr">Nouvelle-Zélande</country><wicri:regionArea>Department of Biology, School of Science, University of Waikato, Hamilton 3216</wicri:regionArea><wicri:noRegion>Hamilton 3216</wicri:noRegion></affiliation></author><author><name sortKey="Williams, Nari" sort="Williams, Nari" uniqKey="Williams N" first="Nari" last="Williams">Nari Williams</name><affiliation wicri:level="1"><nlm:affiliation>Forest Protection, Scion, Rotorua 3010, New Zealand.</nlm:affiliation><country xml:lang="fr">Nouvelle-Zélande</country><wicri:regionArea>Forest Protection, Scion, Rotorua 3010</wicri:regionArea><wicri:noRegion>Rotorua 3010</wicri:noRegion></affiliation></author><author><name sortKey="Scott, Peter" sort="Scott, Peter" uniqKey="Scott P" first="Peter" last="Scott">Peter Scott</name><affiliation wicri:level="1"><nlm:affiliation>Forest Protection, Scion, Rotorua 3010, New Zealand. 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Clearwater</name><affiliation wicri:level="1"><nlm:affiliation>Department of Biology, School of Science, University of Waikato, Hamilton 3216, New Zealand.</nlm:affiliation><country xml:lang="fr">Nouvelle-Zélande</country><wicri:regionArea>Department of Biology, School of Science, University of Waikato, Hamilton 3216</wicri:regionArea><wicri:noRegion>Hamilton 3216</wicri:noRegion></affiliation></author><author><name sortKey="Williams, Nari" sort="Williams, Nari" uniqKey="Williams N" first="Nari" last="Williams">Nari Williams</name><affiliation wicri:level="1"><nlm:affiliation>Forest Protection, Scion, Rotorua 3010, New Zealand.</nlm:affiliation><country xml:lang="fr">Nouvelle-Zélande</country><wicri:regionArea>Forest Protection, Scion, Rotorua 3010</wicri:regionArea><wicri:noRegion>Rotorua 3010</wicri:noRegion></affiliation></author><author><name sortKey="Scott, Peter" sort="Scott, Peter" uniqKey="Scott P" first="Peter" last="Scott">Peter Scott</name><affiliation wicri:level="1"><nlm:affiliation>Forest Protection, Scion, Rotorua 3010, New Zealand. Electronic address: peter.scott@scionresearch.com.</nlm:affiliation><country xml:lang="fr">Nouvelle-Zélande</country><wicri:regionArea>Forest Protection, Scion, Rotorua 3010</wicri:regionArea><wicri:noRegion>Rotorua 3010</wicri:noRegion></affiliation></author></analytic><series><title level="j">Journal of microbiological methods</title><idno type="eISSN">1872-8359</idno><imprint><date when="2018" type="published">2018</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Drug Tolerance (MeSH)</term><term>Forests (MeSH)</term><term>Fungicides, Industrial (pharmacology)</term><term>Microbial Sensitivity Tests (methods)</term><term>Mycelium (growth & development)</term><term>New Zealand (MeSH)</term><term>Oomycetes (drug effects)</term><term>Oomycetes (growth & development)</term><term>Phosphites (pharmacology)</term><term>Phytophthora (drug effects)</term><term>Phytophthora (growth & development)</term><term>Plant Diseases (MeSH)</term><term>Plant Roots (microbiology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Fongicides industriels (pharmacologie)</term><term>Forêts (MeSH)</term><term>Maladies des plantes (MeSH)</term><term>Mycelium (croissance et développement)</term><term>Nouvelle-Zélande (MeSH)</term><term>Oomycetes (croissance et développement)</term><term>Oomycetes (effets des médicaments et des substances chimiques)</term><term>Phosphites (pharmacologie)</term><term>Phytophthora (croissance et développement)</term><term>Phytophthora (effets des médicaments et des substances chimiques)</term><term>Racines de plante (microbiologie)</term><term>Tests de sensibilité microbienne (méthodes)</term><term>Tolérance aux médicaments (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Fungicides, Industrial</term><term>Phosphites</term></keywords><keywords scheme="MESH" type="geographic" xml:lang="en"><term>New Zealand</term></keywords><keywords scheme="MESH" qualifier="croissance et développement" xml:lang="fr"><term>Mycelium</term><term>Oomycetes</term><term>Phytophthora</term></keywords><keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>Oomycetes</term><term>Phytophthora</term></keywords><keywords scheme="MESH" qualifier="effets des médicaments et des substances chimiques" xml:lang="fr"><term>Oomycetes</term><term>Phytophthora</term></keywords><keywords scheme="MESH" qualifier="growth & development" xml:lang="en"><term>Mycelium</term><term>Oomycetes</term><term>Phytophthora</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Microbial Sensitivity Tests</term></keywords><keywords scheme="MESH" qualifier="microbiologie" xml:lang="fr"><term>Racines de plante</term></keywords><keywords scheme="MESH" qualifier="microbiology" xml:lang="en"><term>Plant Roots</term></keywords><keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>Tests de sensibilité microbienne</term></keywords><keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Fongicides industriels</term><term>Phosphites</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Drug Tolerance</term><term>Forests</term><term>Plant Diseases</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Forêts</term><term>Maladies des plantes</term><term>Nouvelle-Zélande</term><term>Tolérance aux médicaments</term></keywords><keywords scheme="Wicri" type="geographic" xml:lang="fr"><term>Nouvelle-Zélande</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A high-throughput assay was developed to screen Phytophthora species for fungicide sensitivity using optical density measurements for unbiased, automated measurement of mycelial growth. The efficacy of the optical density assay (OD) to measure phosphite sensitivity in Phytophthora species was compared to two widely used methods, radial growth (RG) and dry weight (DW) assays. Three isolates of each of Phytophthora cinnamomi, P. multivora and P. pluvialis, with known phosphite exposure and three isolates of each species with no prior phosphite exposure, were screened for phosphite sensitivity using the three assays. Mycelial growth measurements were taken after culturing for 6, 14 and 15 days for the OD, DW and RG assays respectively. Mycelial growth inhibition at 15, 80, 200 and 500 μg/mL phosphite relative to growth on control media was used to determine effective concentration values for 50% growth reduction (EC50). The species varied in their tolerance to phosphite with P. cinnamomi being the least sensitive followed by P. multivora and P. pluvialis. No significant differences in tolerance were found between isolates within the same species using any method. The OD assay produced comparable EC50 values to the RG and DW assays. The growth of the three species was more sensitive to phosphite in the DW than the RG and OD assays, however limited sample throughput and greater variation in measuring small amounts of mycelia in the dry weight assessment increase variability and limits throughput. The OD assay offers a fast method to enable an inventory of chemical resistance and is particularly advantageous for slow growing species as it requires less time and offers greater throughput than existing RG and DW methods.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">30308213</PMID><DateCompleted><Year>2019</Year><Month>07</Month><Day>01</Day></DateCompleted><DateRevised><Year>2019</Year><Month>07</Month><Day>01</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1872-8359</ISSN><JournalIssue CitedMedium="Internet"><Volume>154</Volume><PubDate><Year>2018</Year><Month>11</Month></PubDate></JournalIssue><Title>Journal of microbiological methods</Title><ISOAbbreviation>J Microbiol Methods</ISOAbbreviation></Journal><ArticleTitle>Development of a high throughput optical density assay to determine fungicide sensitivity of oomycetes.</ArticleTitle><Pagination><MedlinePgn>33-39</MedlinePgn></Pagination><ELocationID EIdType="pii" ValidYN="Y">S0167-7012(18)30495-0</ELocationID><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.mimet.2018.10.006</ELocationID><Abstract><AbstractText>A high-throughput assay was developed to screen Phytophthora species for fungicide sensitivity using optical density measurements for unbiased, automated measurement of mycelial growth. The efficacy of the optical density assay (OD) to measure phosphite sensitivity in Phytophthora species was compared to two widely used methods, radial growth (RG) and dry weight (DW) assays. Three isolates of each of Phytophthora cinnamomi, P. multivora and P. pluvialis, with known phosphite exposure and three isolates of each species with no prior phosphite exposure, were screened for phosphite sensitivity using the three assays. Mycelial growth measurements were taken after culturing for 6, 14 and 15 days for the OD, DW and RG assays respectively. Mycelial growth inhibition at 15, 80, 200 and 500 μg/mL phosphite relative to growth on control media was used to determine effective concentration values for 50% growth reduction (EC50). The species varied in their tolerance to phosphite with P. cinnamomi being the least sensitive followed by P. multivora and P. pluvialis. No significant differences in tolerance were found between isolates within the same species using any method. The OD assay produced comparable EC50 values to the RG and DW assays. The growth of the three species was more sensitive to phosphite in the DW than the RG and OD assays, however limited sample throughput and greater variation in measuring small amounts of mycelia in the dry weight assessment increase variability and limits throughput. The OD assay offers a fast method to enable an inventory of chemical resistance and is particularly advantageous for slow growing species as it requires less time and offers greater throughput than existing RG and DW methods.</AbstractText><CopyrightInformation>Copyright © 2018. Published by Elsevier B.V.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Hunter</LastName><ForeName>Shannon</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Department of Biology, School of Science, University of Waikato, Hamilton 3216, New Zealand; Forest Protection, Scion, Rotorua 3010, New Zealand.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>McDougal</LastName><ForeName>Rebecca</ForeName><Initials>R</Initials><AffiliationInfo><Affiliation>Forest Protection, Scion, Rotorua 3010, New Zealand.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Clearwater</LastName><ForeName>Mike J</ForeName><Initials>MJ</Initials><AffiliationInfo><Affiliation>Department of Biology, School of Science, University of Waikato, Hamilton 3216, New Zealand.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Williams</LastName><ForeName>Nari</ForeName><Initials>N</Initials><AffiliationInfo><Affiliation>Forest Protection, Scion, Rotorua 3010, New Zealand.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Scott</LastName><ForeName>Peter</ForeName><Initials>P</Initials><AffiliationInfo><Affiliation>Forest Protection, Scion, Rotorua 3010, New Zealand. Electronic address: peter.scott@scionresearch.com.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2018</Year><Month>10</Month><Day>09</Day></ArticleDate></Article><MedlineJournalInfo><Country>Netherlands</Country><MedlineTA>J Microbiol Methods</MedlineTA><NlmUniqueID>8306883</NlmUniqueID><ISSNLinking>0167-7012</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D005659">Fungicides, Industrial</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D017905">Phosphites</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D004361" MajorTopicYN="N">Drug Tolerance</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D065928" MajorTopicYN="N">Forests</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D005659" MajorTopicYN="N">Fungicides, Industrial</DescriptorName><QualifierName UI="Q000494" MajorTopicYN="Y">pharmacology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D008826" MajorTopicYN="N">Microbial Sensitivity Tests</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D025282" MajorTopicYN="N">Mycelium</DescriptorName><QualifierName UI="Q000254" MajorTopicYN="N">growth & development</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D009520" MajorTopicYN="N" Type="Geographic">New Zealand</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D009868" MajorTopicYN="N">Oomycetes</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="Y">drug effects</QualifierName><QualifierName UI="Q000254" MajorTopicYN="Y">growth & development</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D017905" MajorTopicYN="N">Phosphites</DescriptorName><QualifierName UI="Q000494" MajorTopicYN="N">pharmacology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010838" MajorTopicYN="N">Phytophthora</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="N">drug effects</QualifierName><QualifierName UI="Q000254" MajorTopicYN="N">growth & development</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010935" MajorTopicYN="N">Plant Diseases</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018517" MajorTopicYN="N">Plant Roots</DescriptorName><QualifierName UI="Q000382" MajorTopicYN="N">microbiology</QualifierName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="Y">Fungicide</Keyword><Keyword MajorTopicYN="Y">Oomycete</Keyword><Keyword MajorTopicYN="Y">Optical density</Keyword><Keyword MajorTopicYN="Y">Phosphite</Keyword><Keyword MajorTopicYN="Y">Phytophthora</Keyword><Keyword MajorTopicYN="Y">Tolerance</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2018</Year><Month>06</Month><Day>21</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2018</Year><Month>09</Month><Day>26</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2018</Year><Month>10</Month><Day>08</Day></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2018</Year><Month>10</Month><Day>12</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2019</Year><Month>7</Month><Day>2</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2018</Year><Month>10</Month><Day>12</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">30308213</ArticleId><ArticleId IdType="pii">S0167-7012(18)30495-0</ArticleId><ArticleId IdType="doi">10.1016/j.mimet.2018.10.006</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Nouvelle-Zélande</li></country></list><tree><country name="Nouvelle-Zélande"><noRegion><name sortKey="Hunter, Shannon" sort="Hunter, Shannon" uniqKey="Hunter S" first="Shannon" last="Hunter">Shannon Hunter</name></noRegion><name sortKey="Clearwater, Mike J" sort="Clearwater, Mike J" uniqKey="Clearwater M" first="Mike J" last="Clearwater">Mike J. Clearwater</name><name sortKey="Mcdougal, Rebecca" sort="Mcdougal, Rebecca" uniqKey="Mcdougal R" first="Rebecca" last="Mcdougal">Rebecca Mcdougal</name><name sortKey="Scott, Peter" sort="Scott, Peter" uniqKey="Scott P" first="Peter" last="Scott">Peter Scott</name><name sortKey="Williams, Nari" sort="Williams, Nari" uniqKey="Williams N" first="Nari" last="Williams">Nari Williams</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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