Isolation of a species-specific mitochondrial DNA sequence for identification of Tilletia indica, the karnal bunt of wheat fungus
Mitochondrial DNA (mtDNA) from five isolates of Tilletia indica was isolated and digested with several restriction enzymes. A 2.3-kb EcoRI fragment was chosen, cloned, and shown to hybridize with total DNA restricted with EcoRI from T. indica and not from a morphologically similar smut fungus, Tille...
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| Veröffentlicht in: | Applied and Environmental Microbiology 1996-01, Vol.62 (1), p.87-93 |
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| creator | Ferreira, M.A.S.V. (Universidade de Brasilia, Brasilia, Brazil.) Tooley, P.W Hatziloukas, E Castro, C Schaad, N.W |
| description | Mitochondrial DNA (mtDNA) from five isolates of Tilletia indica was isolated and digested with several restriction enzymes. A 2.3-kb EcoRI fragment was chosen, cloned, and shown to hybridize with total DNA restricted with EcoRI from T. indica and not from a morphologically similar smut fungus, Tilletia barclayana. The clone was partially sequenced, and primers were designed and tested under high-stringency conditions in PCR assays. The primer pair Ti1/Ti4 amplified a 2.3-kb fragment from total DNA of 17 T. indica isolates from India, Pakistan, and Mexico. DNA from 25 isolates of other smut fungi (T. barclayana, Tilletia foetida, Tilletia caries, Tilletia fusca, and Tilletia controversa) did not produce any bands, as detected by ethidium bromide-stained agarose gels and Southern hybridizations. The sensitivity of the assay was determined and increased by using a single nested primer in a second round of amplification, so that 1 pg of total mycelial DNA could be detected. The results indicated that the primers which originated from a cloned mtDNA sequence can be used to differentiate T. indica from other Tilletia species and have the potential to identify teliospores contaminating wheat seeds |
| doi_str_mv | 10.1128/aem.62.1.87-93.1996 |
| format | Article |
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(Universidade de Brasilia, Brasilia, Brazil.) ; Tooley, P.W ; Hatziloukas, E ; Castro, C ; Schaad, N.W</creator><creatorcontrib>Ferreira, M.A.S.V. (Universidade de Brasilia, Brasilia, Brazil.) ; Tooley, P.W ; Hatziloukas, E ; Castro, C ; Schaad, N.W</creatorcontrib><description>Mitochondrial DNA (mtDNA) from five isolates of Tilletia indica was isolated and digested with several restriction enzymes. A 2.3-kb EcoRI fragment was chosen, cloned, and shown to hybridize with total DNA restricted with EcoRI from T. indica and not from a morphologically similar smut fungus, Tilletia barclayana. The clone was partially sequenced, and primers were designed and tested under high-stringency conditions in PCR assays. The primer pair Ti1/Ti4 amplified a 2.3-kb fragment from total DNA of 17 T. indica isolates from India, Pakistan, and Mexico. DNA from 25 isolates of other smut fungi (T. barclayana, Tilletia foetida, Tilletia caries, Tilletia fusca, and Tilletia controversa) did not produce any bands, as detected by ethidium bromide-stained agarose gels and Southern hybridizations. The sensitivity of the assay was determined and increased by using a single nested primer in a second round of amplification, so that 1 pg of total mycelial DNA could be detected. The results indicated that the primers which originated from a cloned mtDNA sequence can be used to differentiate T. indica from other Tilletia species and have the potential to identify teliospores contaminating wheat seeds</description><identifier>ISSN: 0099-2240</identifier><identifier>EISSN: 1098-5336</identifier><identifier>DOI: 10.1128/aem.62.1.87-93.1996</identifier><identifier>PMID: 8572716</identifier><identifier>CODEN: AEMIDF</identifier><language>eng</language><publisher>Washington, DC: American Society for Microbiology</publisher><subject>ADN ; AMPLIFICATION CHAINE POLYMERASE ; Base Sequence ; Biological and medical sciences ; Cellular biology ; CHAMPIGNON ; Cloning, Molecular ; Deoxyribonucleic acid ; DNA ; DNA Primers ; DNA, Fungal - analysis ; DNA, Fungal - genetics ; DNA, Fungal - isolation & purification ; DNA, Mitochondrial - analysis ; DNA, Mitochondrial - genetics ; DNA, Mitochondrial - isolation & purification ; Enzymes ; ESPORAS ; Fundamental and applied biological sciences. Psychology ; Fungi ; HIBRIDACION DE ADN ; HONGOS ; HYBRIDATION D'ADN ; IDENTIFICACION ; IDENTIFICATION ; Microbiology ; MITOCHONDRIE ; MITOCONDRIA ; Molecular Sequence Data ; Mycological methods and techniques used in mycology ; Mycology ; ORGANISME TRANSMISSIBLE PAR SEMENCE ; ORGANISMOS TRANSMITIDOS POR SEMILLA ; Polymerase Chain Reaction - methods ; REACCION DE CADENAS DE POLIMERASA ; SECUENCIA NUCLEICA ; Sensitivity and Specificity ; Sequence Analysis, DNA ; SEQUENCE NUCLEIQUE ; Species Specificity ; SPORE ; TECHNIQUE ANALYTIQUE ; TECNICAS ANALITICAS ; TILLETIA ; Tilletia barclayana ; TILLETIA INDICA ; Triticum - microbiology ; TRITICUM AESTIVUM ; Ustilaginales - genetics ; Wheat</subject><ispartof>Applied and Environmental Microbiology, 1996-01, Vol.62 (1), p.87-93</ispartof><rights>1996 INIST-CNRS</rights><rights>Copyright American Society for Microbiology Jan 1996</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c615t-1c19655315cf2f97d4d69914e1081ddb0ac50bf9b8df11c3036eee1d83790b053</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC167776/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC167776/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,315,728,781,785,886,3189,3190,4025,27928,27929,27930,53796,53798</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=3101519$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8572716$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ferreira, M.A.S.V. (Universidade de Brasilia, Brasilia, Brazil.)</creatorcontrib><creatorcontrib>Tooley, P.W</creatorcontrib><creatorcontrib>Hatziloukas, E</creatorcontrib><creatorcontrib>Castro, C</creatorcontrib><creatorcontrib>Schaad, N.W</creatorcontrib><title>Isolation of a species-specific mitochondrial DNA sequence for identification of Tilletia indica, the karnal bunt of wheat fungus</title><title>Applied and Environmental Microbiology</title><addtitle>Appl Environ Microbiol</addtitle><description>Mitochondrial DNA (mtDNA) from five isolates of Tilletia indica was isolated and digested with several restriction enzymes. A 2.3-kb EcoRI fragment was chosen, cloned, and shown to hybridize with total DNA restricted with EcoRI from T. indica and not from a morphologically similar smut fungus, Tilletia barclayana. The clone was partially sequenced, and primers were designed and tested under high-stringency conditions in PCR assays. The primer pair Ti1/Ti4 amplified a 2.3-kb fragment from total DNA of 17 T. indica isolates from India, Pakistan, and Mexico. DNA from 25 isolates of other smut fungi (T. barclayana, Tilletia foetida, Tilletia caries, Tilletia fusca, and Tilletia controversa) did not produce any bands, as detected by ethidium bromide-stained agarose gels and Southern hybridizations. The sensitivity of the assay was determined and increased by using a single nested primer in a second round of amplification, so that 1 pg of total mycelial DNA could be detected. The results indicated that the primers which originated from a cloned mtDNA sequence can be used to differentiate T. indica from other Tilletia species and have the potential to identify teliospores contaminating wheat seeds</description><subject>ADN</subject><subject>AMPLIFICATION CHAINE POLYMERASE</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Cellular biology</subject><subject>CHAMPIGNON</subject><subject>Cloning, Molecular</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA Primers</subject><subject>DNA, Fungal - analysis</subject><subject>DNA, Fungal - genetics</subject><subject>DNA, Fungal - isolation & purification</subject><subject>DNA, Mitochondrial - analysis</subject><subject>DNA, Mitochondrial - genetics</subject><subject>DNA, Mitochondrial - isolation & purification</subject><subject>Enzymes</subject><subject>ESPORAS</subject><subject>Fundamental and applied biological sciences. 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(Universidade de Brasilia, Brasilia, Brazil.)</creator><creator>Tooley, P.W</creator><creator>Hatziloukas, E</creator><creator>Castro, C</creator><creator>Schaad, N.W</creator><general>American Society for Microbiology</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QO</scope><scope>7SN</scope><scope>7SS</scope><scope>7ST</scope><scope>7T7</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>SOI</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>199601</creationdate><title>Isolation of a species-specific mitochondrial DNA sequence for identification of Tilletia indica, the karnal bunt of wheat fungus</title><author>Ferreira, M.A.S.V. (Universidade de Brasilia, Brasilia, Brazil.) ; Tooley, P.W ; Hatziloukas, E ; Castro, C ; Schaad, N.W</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c615t-1c19655315cf2f97d4d69914e1081ddb0ac50bf9b8df11c3036eee1d83790b053</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>ADN</topic><topic>AMPLIFICATION CHAINE POLYMERASE</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Cellular biology</topic><topic>CHAMPIGNON</topic><topic>Cloning, Molecular</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA Primers</topic><topic>DNA, Fungal - analysis</topic><topic>DNA, Fungal - genetics</topic><topic>DNA, Fungal - isolation & purification</topic><topic>DNA, Mitochondrial - analysis</topic><topic>DNA, Mitochondrial - genetics</topic><topic>DNA, Mitochondrial - isolation & purification</topic><topic>Enzymes</topic><topic>ESPORAS</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fungi</topic><topic>HIBRIDACION DE ADN</topic><topic>HONGOS</topic><topic>HYBRIDATION D'ADN</topic><topic>IDENTIFICACION</topic><topic>IDENTIFICATION</topic><topic>Microbiology</topic><topic>MITOCHONDRIE</topic><topic>MITOCONDRIA</topic><topic>Molecular Sequence Data</topic><topic>Mycological methods and techniques used in mycology</topic><topic>Mycology</topic><topic>ORGANISME TRANSMISSIBLE PAR SEMENCE</topic><topic>ORGANISMOS TRANSMITIDOS POR SEMILLA</topic><topic>Polymerase Chain Reaction - methods</topic><topic>REACCION DE CADENAS DE POLIMERASA</topic><topic>SECUENCIA NUCLEICA</topic><topic>Sensitivity and Specificity</topic><topic>Sequence Analysis, DNA</topic><topic>SEQUENCE NUCLEIQUE</topic><topic>Species Specificity</topic><topic>SPORE</topic><topic>TECHNIQUE ANALYTIQUE</topic><topic>TECNICAS ANALITICAS</topic><topic>TILLETIA</topic><topic>Tilletia barclayana</topic><topic>TILLETIA INDICA</topic><topic>Triticum - microbiology</topic><topic>TRITICUM AESTIVUM</topic><topic>Ustilaginales - genetics</topic><topic>Wheat</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ferreira, M.A.S.V. (Universidade de Brasilia, Brasilia, Brazil.)</creatorcontrib><creatorcontrib>Tooley, P.W</creatorcontrib><creatorcontrib>Hatziloukas, E</creatorcontrib><creatorcontrib>Castro, C</creatorcontrib><creatorcontrib>Schaad, N.W</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Environment Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>Environment Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Applied and Environmental Microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ferreira, M.A.S.V. (Universidade de Brasilia, Brasilia, Brazil.)</au><au>Tooley, P.W</au><au>Hatziloukas, E</au><au>Castro, C</au><au>Schaad, N.W</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Isolation of a species-specific mitochondrial DNA sequence for identification of Tilletia indica, the karnal bunt of wheat fungus</atitle><jtitle>Applied and Environmental Microbiology</jtitle><addtitle>Appl Environ Microbiol</addtitle><date>1996-01</date><risdate>1996</risdate><volume>62</volume><issue>1</issue><spage>87</spage><epage>93</epage><pages>87-93</pages><issn>0099-2240</issn><eissn>1098-5336</eissn><coden>AEMIDF</coden><abstract>Mitochondrial DNA (mtDNA) from five isolates of Tilletia indica was isolated and digested with several restriction enzymes. A 2.3-kb EcoRI fragment was chosen, cloned, and shown to hybridize with total DNA restricted with EcoRI from T. indica and not from a morphologically similar smut fungus, Tilletia barclayana. The clone was partially sequenced, and primers were designed and tested under high-stringency conditions in PCR assays. The primer pair Ti1/Ti4 amplified a 2.3-kb fragment from total DNA of 17 T. indica isolates from India, Pakistan, and Mexico. DNA from 25 isolates of other smut fungi (T. barclayana, Tilletia foetida, Tilletia caries, Tilletia fusca, and Tilletia controversa) did not produce any bands, as detected by ethidium bromide-stained agarose gels and Southern hybridizations. The sensitivity of the assay was determined and increased by using a single nested primer in a second round of amplification, so that 1 pg of total mycelial DNA could be detected. The results indicated that the primers which originated from a cloned mtDNA sequence can be used to differentiate T. indica from other Tilletia species and have the potential to identify teliospores contaminating wheat seeds</abstract><cop>Washington, DC</cop><pub>American Society for Microbiology</pub><pmid>8572716</pmid><doi>10.1128/aem.62.1.87-93.1996</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0099-2240 |
| ispartof | Applied and Environmental Microbiology, 1996-01, Vol.62 (1), p.87-93 |
| issn | 0099-2240 1098-5336 |
| language | eng |
| recordid | cdi_pubmed_primary_8572716 |
| source | MEDLINE; American Society for Microbiology Journals; PubMed Central; Alma/SFX Local Collection |
| subjects | ADN AMPLIFICATION CHAINE POLYMERASE Base Sequence Biological and medical sciences Cellular biology CHAMPIGNON Cloning, Molecular Deoxyribonucleic acid DNA DNA Primers DNA, Fungal - analysis DNA, Fungal - genetics DNA, Fungal - isolation & purification DNA, Mitochondrial - analysis DNA, Mitochondrial - genetics DNA, Mitochondrial - isolation & purification Enzymes ESPORAS Fundamental and applied biological sciences. Psychology Fungi HIBRIDACION DE ADN HONGOS HYBRIDATION D'ADN IDENTIFICACION IDENTIFICATION Microbiology MITOCHONDRIE MITOCONDRIA Molecular Sequence Data Mycological methods and techniques used in mycology Mycology ORGANISME TRANSMISSIBLE PAR SEMENCE ORGANISMOS TRANSMITIDOS POR SEMILLA Polymerase Chain Reaction - methods REACCION DE CADENAS DE POLIMERASA SECUENCIA NUCLEICA Sensitivity and Specificity Sequence Analysis, DNA SEQUENCE NUCLEIQUE Species Specificity SPORE TECHNIQUE ANALYTIQUE TECNICAS ANALITICAS TILLETIA Tilletia barclayana TILLETIA INDICA Triticum - microbiology TRITICUM AESTIVUM Ustilaginales - genetics Wheat |
| title | Isolation of a species-specific mitochondrial DNA sequence for identification of Tilletia indica, the karnal bunt of wheat fungus |
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