Direct colony PCR-SSCP for detection of multiple pythiaceous oomycetes in environmental samples.
Identifieur interne : 002310 ( Main/Corpus ); précédent : 002309; suivant : 002311Direct colony PCR-SSCP for detection of multiple pythiaceous oomycetes in environmental samples.
Auteurs : Ping Kong ; Patricia A. Richardson ; Chuanxue HongSource :
- Journal of microbiological methods [ 0167-7012 ] ; 2005.
English descriptors
- KwdEn :
- DNA, Fungal (chemistry), DNA, Fungal (genetics), Daphne (microbiology), Lycopersicon esculentum (microbiology), Phytophthora (genetics), Phytophthora (isolation & purification), Plant Diseases (microbiology), Polymerase Chain Reaction (methods), Polymorphism, Single-Stranded Conformational (MeSH), Pythium (genetics), Pythium (isolation & purification), Rhododendron (microbiology), Soil Microbiology (MeSH), Tobacco (microbiology), Vinca (microbiology).
- MESH :
- chemical , chemistry : DNA, Fungal.
- chemical , genetics : DNA, Fungal.
- genetics : Phytophthora, Pythium.
- isolation & purification : Phytophthora, Pythium.
- methods : Polymerase Chain Reaction.
- microbiology : Daphne, Lycopersicon esculentum, Plant Diseases, Rhododendron, Tobacco, Vinca.
- Polymorphism, Single-Stranded Conformational, Soil Microbiology.
Abstract
Colony PCR was developed for detection of pythiaceous species recovered on selective agar plates without DNA extraction. A minute amount of mycelia from a single colony was picked up with a pipette tip and added directly to the PCR mix as template for DNA amplification. Successful amplification was achieved in over 95% of the colonies recovered from plant tissues, irrigation water and soil with species-specific primers or oomycete ITS-1 primers. PCR was inhibited in the case of colonies emerging from unwashed pine bark potting mix plates. Direct colony PCR with ITS-1 primers combined with single-strand conformation polymorphism analysis (SSCP) was used to determine population levels of single and multiple species in plant and environmental samples. Application of this technique for disease diagnosis and monitoring pathogen sources was explored, and the potential for studying diversity and population dynamics of other cultivated microbial communities in the environment is discussed.
DOI: 10.1016/j.mimet.2004.10.019
PubMed: 15676193
Links to Exploration step
pubmed:15676193Le document en format XML
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Richardson</name></author><author><name sortKey="Hong, Chuanxue" sort="Hong, Chuanxue" uniqKey="Hong C" first="Chuanxue" last="Hong">Chuanxue Hong</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2005">2005</date><idno type="RBID">pubmed:15676193</idno><idno type="pmid">15676193</idno><idno type="doi">10.1016/j.mimet.2004.10.019</idno><idno type="wicri:Area/Main/Corpus">002310</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">002310</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Direct colony PCR-SSCP for detection of multiple pythiaceous oomycetes in environmental samples.</title><author><name sortKey="Kong, Ping" sort="Kong, Ping" uniqKey="Kong P" first="Ping" last="Kong">Ping Kong</name><affiliation><nlm:affiliation>Department of Plant Pathology, Physiology, and Weed Science, Virginia Polytechnic Institute and State University, Hampton Roads Agricultural Research and Extension Center, 1444 Diamond Springs Road, Virginia Beach, VA 23455, USA. pkong@vt.edu</nlm:affiliation></affiliation></author><author><name sortKey="Richardson, Patricia A" sort="Richardson, Patricia A" uniqKey="Richardson P" first="Patricia A" last="Richardson">Patricia A. Richardson</name></author><author><name sortKey="Hong, Chuanxue" sort="Hong, Chuanxue" uniqKey="Hong C" first="Chuanxue" last="Hong">Chuanxue Hong</name></author></analytic><series><title level="j">Journal of microbiological methods</title><idno type="ISSN">0167-7012</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>DNA, Fungal (chemistry)</term><term>DNA, Fungal (genetics)</term><term>Daphne (microbiology)</term><term>Lycopersicon esculentum (microbiology)</term><term>Phytophthora (genetics)</term><term>Phytophthora (isolation & purification)</term><term>Plant Diseases (microbiology)</term><term>Polymerase Chain Reaction (methods)</term><term>Polymorphism, Single-Stranded Conformational (MeSH)</term><term>Pythium (genetics)</term><term>Pythium (isolation & purification)</term><term>Rhododendron (microbiology)</term><term>Soil Microbiology (MeSH)</term><term>Tobacco (microbiology)</term><term>Vinca (microbiology)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>DNA, Fungal</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA, Fungal</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Phytophthora</term><term>Pythium</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Phytophthora</term><term>Pythium</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" qualifier="microbiology" xml:lang="en"><term>Daphne</term><term>Lycopersicon esculentum</term><term>Plant Diseases</term><term>Rhododendron</term><term>Tobacco</term><term>Vinca</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Polymorphism, Single-Stranded Conformational</term><term>Soil Microbiology</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Colony PCR was developed for detection of pythiaceous species recovered on selective agar plates without DNA extraction. A minute amount of mycelia from a single colony was picked up with a pipette tip and added directly to the PCR mix as template for DNA amplification. Successful amplification was achieved in over 95% of the colonies recovered from plant tissues, irrigation water and soil with species-specific primers or oomycete ITS-1 primers. PCR was inhibited in the case of colonies emerging from unwashed pine bark potting mix plates. Direct colony PCR with ITS-1 primers combined with single-strand conformation polymorphism analysis (SSCP) was used to determine population levels of single and multiple species in plant and environmental samples. Application of this technique for disease diagnosis and monitoring pathogen sources was explored, and the potential for studying diversity and population dynamics of other cultivated microbial communities in the environment is discussed.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">15676193</PMID><DateCompleted><Year>2005</Year><Month>04</Month><Day>05</Day></DateCompleted><DateRevised><Year>2005</Year><Month>01</Month><Day>28</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0167-7012</ISSN><JournalIssue CitedMedium="Print"><Volume>61</Volume><Issue>1</Issue><PubDate><Year>2005</Year><Month>Apr</Month></PubDate></JournalIssue><Title>Journal of microbiological methods</Title><ISOAbbreviation>J Microbiol Methods</ISOAbbreviation></Journal><ArticleTitle>Direct colony PCR-SSCP for detection of multiple pythiaceous oomycetes in environmental samples.</ArticleTitle><Pagination><MedlinePgn>25-32</MedlinePgn></Pagination><Abstract><AbstractText>Colony PCR was developed for detection of pythiaceous species recovered on selective agar plates without DNA extraction. A minute amount of mycelia from a single colony was picked up with a pipette tip and added directly to the PCR mix as template for DNA amplification. Successful amplification was achieved in over 95% of the colonies recovered from plant tissues, irrigation water and soil with species-specific primers or oomycete ITS-1 primers. PCR was inhibited in the case of colonies emerging from unwashed pine bark potting mix plates. Direct colony PCR with ITS-1 primers combined with single-strand conformation polymorphism analysis (SSCP) was used to determine population levels of single and multiple species in plant and environmental samples. Application of this technique for disease diagnosis and monitoring pathogen sources was explored, and the potential for studying diversity and population dynamics of other cultivated microbial communities in the environment is discussed.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Kong</LastName><ForeName>Ping</ForeName><Initials>P</Initials><AffiliationInfo><Affiliation>Department of Plant Pathology, Physiology, and Weed Science, Virginia Polytechnic Institute and State University, Hampton Roads Agricultural Research and Extension Center, 1444 Diamond Springs Road, Virginia Beach, VA 23455, USA. pkong@vt.edu</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Richardson</LastName><ForeName>Patricia A</ForeName><Initials>PA</Initials></Author><Author ValidYN="Y"><LastName>Hong</LastName><ForeName>Chuanxue</ForeName><Initials>C</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList></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="D004271">DNA, Fungal</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D004271" MajorTopicYN="N">DNA, Fungal</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D032364" MajorTopicYN="N">Daphne</DescriptorName><QualifierName UI="Q000382" MajorTopicYN="N">microbiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018551" MajorTopicYN="N">Lycopersicon esculentum</DescriptorName><QualifierName UI="Q000382" MajorTopicYN="N">microbiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010838" MajorTopicYN="N">Phytophthora</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010935" MajorTopicYN="N">Plant Diseases</DescriptorName><QualifierName UI="Q000382" MajorTopicYN="Y">microbiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D016133" MajorTopicYN="N">Polymerase Chain Reaction</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018807" MajorTopicYN="N">Polymorphism, Single-Stranded Conformational</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D011775" MajorTopicYN="N">Pythium</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D029793" MajorTopicYN="N">Rhododendron</DescriptorName><QualifierName UI="Q000382" MajorTopicYN="N">microbiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D012988" MajorTopicYN="N">Soil Microbiology</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D014026" MajorTopicYN="N">Tobacco</DescriptorName><QualifierName UI="Q000382" MajorTopicYN="N">microbiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D031053" MajorTopicYN="N">Vinca</DescriptorName><QualifierName UI="Q000382" MajorTopicYN="N">microbiology</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2004</Year><Month>07</Month><Day>02</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2004</Year><Month>10</Month><Day>26</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2004</Year><Month>10</Month><Day>27</Day></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2005</Year><Month>1</Month><Day>29</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2005</Year><Month>4</Month><Day>6</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2005</Year><Month>1</Month><Day>29</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">15676193</ArticleId><ArticleId IdType="pii">S0167-7012(04)00302-1</ArticleId><ArticleId IdType="doi">10.1016/j.mimet.2004.10.019</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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