Quantification of conidial density of Aspergillus flavus and A. parasiticus in soil from almond orchards using real-time PCR
To design the Aspergillus flavus and Aspergillus parasiticus-specific primers and a real-time PCR assay for quantification of the conidial density in soil. Aspergillus flavus and A. parasiticus-specific DNA primers were designed based on internal transcribed spacer sequences to distinguish these two...
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| Veröffentlicht in: | Journal of applied microbiology 2009-05, Vol.106 (5), p.1649-1660 |
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| creator | Luo, Y Gao, W Doster, M Michailides, T.J |
| description | To design the Aspergillus flavus and Aspergillus parasiticus-specific primers and a real-time PCR assay for quantification of the conidial density in soil. Aspergillus flavus and A. parasiticus-specific DNA primers were designed based on internal transcribed spacer sequences to distinguish these two species and from other Aspergillus and other fungal species. A method of pathogen DNA extraction directly from soil samples was developed. Using the designed primers, a real-time PCR assay was developed to quantitatively determine the conidial density of each A. flavus and A. parasiticus in soil, after generating corresponding standard curves. Known conidial densities of each A. flavus or A. parasiticus in soil significantly correlated with those tested with the real-time PCR. This study demonstrated the applicability of the real-time PCR assay in studies of quantifying A. flavus and A. parasiticus in soil as inoculum sources. The A. flacus and A. parasitic-specific primers can be widely used in aflatoxin research. The real-time PCR assay developed in this study provides a potential approach to quantify the plant pathogen density from not only soil but also other sources in relation to aflatoxin contamination from environment, food and feed commodities. |
| doi_str_mv | 10.1111/j.1365-2672.2008.04132.x |
| format | Article |
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Aspergillus flavus and A. parasiticus-specific DNA primers were designed based on internal transcribed spacer sequences to distinguish these two species and from other Aspergillus and other fungal species. A method of pathogen DNA extraction directly from soil samples was developed. Using the designed primers, a real-time PCR assay was developed to quantitatively determine the conidial density of each A. flavus and A. parasiticus in soil, after generating corresponding standard curves. Known conidial densities of each A. flavus or A. parasiticus in soil significantly correlated with those tested with the real-time PCR. This study demonstrated the applicability of the real-time PCR assay in studies of quantifying A. flavus and A. parasiticus in soil as inoculum sources. The A. flacus and A. parasitic-specific primers can be widely used in aflatoxin research. The real-time PCR assay developed in this study provides a potential approach to quantify the plant pathogen density from not only soil but also other sources in relation to aflatoxin contamination from environment, food and feed commodities.</description><identifier>ISSN: 1364-5072</identifier><identifier>EISSN: 1365-2672</identifier><identifier>DOI: 10.1111/j.1365-2672.2008.04132.x</identifier><identifier>PMID: 19226388</identifier><language>eng</language><publisher>Oxford, UK: Oxford, UK : Blackwell Publishing Ltd</publisher><subject>aflatoxin ; aflatoxins ; Agriculture ; Aspergillus - classification ; Aspergillus - genetics ; Aspergillus - growth & development ; Aspergillus - isolation & purification ; Aspergillus flavus ; Aspergillus parasiticus ; Biological and medical sciences ; DNA Primers - chemistry ; DNA, Fungal - genetics ; DNA, Fungal - isolation & purification ; DNA, Intergenic ; Fundamental and applied biological sciences. Psychology ; fungi ; internal transcribed spacer ; internal transcribed spacers ; Microbiology ; Phylogeny ; Polymerase Chain Reaction - methods ; Prunus ; Prunus dulcis ; quantification ; quantitative analysis ; real-time PCR ; Sequence Alignment ; soil ; Soil Microbiology ; Spores, Fungal - growth & development ; Spores, Fungal - isolation & purification</subject><ispartof>Journal of applied microbiology, 2009-05, Vol.106 (5), p.1649-1660</ispartof><rights>2009 The Authors. Journal compilation © 2009 The Society for Applied Microbiology</rights><rights>2009 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4532-c7aace96886706faf22ceee67627b1c7c3f75f424609b22e2c160db38f402ba73</citedby><cites>FETCH-LOGICAL-c4532-c7aace96886706faf22ceee67627b1c7c3f75f424609b22e2c160db38f402ba73</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fj.1365-2672.2008.04132.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fj.1365-2672.2008.04132.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>315,781,785,1418,27929,27930,45579,45580</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=21345277$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19226388$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Luo, Y</creatorcontrib><creatorcontrib>Gao, W</creatorcontrib><creatorcontrib>Doster, M</creatorcontrib><creatorcontrib>Michailides, T.J</creatorcontrib><title>Quantification of conidial density of Aspergillus flavus and A. parasiticus in soil from almond orchards using real-time PCR</title><title>Journal of applied microbiology</title><addtitle>J Appl Microbiol</addtitle><description>To design the Aspergillus flavus and Aspergillus parasiticus-specific primers and a real-time PCR assay for quantification of the conidial density in soil. Aspergillus flavus and A. parasiticus-specific DNA primers were designed based on internal transcribed spacer sequences to distinguish these two species and from other Aspergillus and other fungal species. A method of pathogen DNA extraction directly from soil samples was developed. Using the designed primers, a real-time PCR assay was developed to quantitatively determine the conidial density of each A. flavus and A. parasiticus in soil, after generating corresponding standard curves. Known conidial densities of each A. flavus or A. parasiticus in soil significantly correlated with those tested with the real-time PCR. This study demonstrated the applicability of the real-time PCR assay in studies of quantifying A. flavus and A. parasiticus in soil as inoculum sources. The A. flacus and A. parasitic-specific primers can be widely used in aflatoxin research. The real-time PCR assay developed in this study provides a potential approach to quantify the plant pathogen density from not only soil but also other sources in relation to aflatoxin contamination from environment, food and feed commodities.</description><subject>aflatoxin</subject><subject>aflatoxins</subject><subject>Agriculture</subject><subject>Aspergillus - classification</subject><subject>Aspergillus - genetics</subject><subject>Aspergillus - growth & development</subject><subject>Aspergillus - isolation & purification</subject><subject>Aspergillus flavus</subject><subject>Aspergillus parasiticus</subject><subject>Biological and medical sciences</subject><subject>DNA Primers - chemistry</subject><subject>DNA, Fungal - genetics</subject><subject>DNA, Fungal - isolation & purification</subject><subject>DNA, Intergenic</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>fungi</subject><subject>internal transcribed spacer</subject><subject>internal transcribed spacers</subject><subject>Microbiology</subject><subject>Phylogeny</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Prunus</subject><subject>Prunus dulcis</subject><subject>quantification</subject><subject>quantitative analysis</subject><subject>real-time PCR</subject><subject>Sequence Alignment</subject><subject>soil</subject><subject>Soil Microbiology</subject><subject>Spores, Fungal - growth & development</subject><subject>Spores, Fungal - isolation & purification</subject><issn>1364-5072</issn><issn>1365-2672</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkk2P0zAQhiMEYpeFvwC-AKcEe5zY6YFDVfGpRXzu2Zo4dnHlxMVOYCvx43G21XJD-DKj8fPO2H5dFITRiuX1YlcxLpoShIQKKG0rWjMO1fWd4vx24-5NXpcNlXBWPEhpRynjtBH3izO2AhC8bc-L359nHCdnncbJhZEES3QYXe_Qk96MyU2HpbZOexO3zvs5EevxZw449mRdkT1GzJTTueRGkoLzxMYwEPRDyEiI-jvGPpE5uXFLokFfTm4w5NPmy8PinkWfzKNTvCiuXr_6tnlbXn58826zvix13XAotUTUZiXaVkgqLFoAbYwRUoDsmJaaW9nYGmpBVx2AAc0E7Tve2ppCh5JfFM-Pffcx_JhNmtTgkjbe42jCnJTknK44gzqTz_5JAm0kE_Uqg-0R1DGkFI1V--gGjAfFqFo8Uju1WKEWK9TikbrxSF1n6ePTjLkbTP9XeDIlA09PACaN3kYctUu3HDBeNyCXa708cr-cN4f_PoB6v_6wZFn_5Ki3GBRuY55x9RWWT8IEEyK__R8FybaP</recordid><startdate>200905</startdate><enddate>200905</enddate><creator>Luo, Y</creator><creator>Gao, W</creator><creator>Doster, M</creator><creator>Michailides, T.J</creator><general>Oxford, UK : Blackwell Publishing Ltd</general><general>Blackwell Publishing Ltd</general><general>Blackwell</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>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>200905</creationdate><title>Quantification of conidial density of Aspergillus flavus and A. parasiticus in soil from almond orchards using real-time PCR</title><author>Luo, Y ; Gao, W ; Doster, M ; Michailides, T.J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4532-c7aace96886706faf22ceee67627b1c7c3f75f424609b22e2c160db38f402ba73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>aflatoxin</topic><topic>aflatoxins</topic><topic>Agriculture</topic><topic>Aspergillus - classification</topic><topic>Aspergillus - genetics</topic><topic>Aspergillus - growth & development</topic><topic>Aspergillus - isolation & purification</topic><topic>Aspergillus flavus</topic><topic>Aspergillus parasiticus</topic><topic>Biological and medical sciences</topic><topic>DNA Primers - chemistry</topic><topic>DNA, Fungal - genetics</topic><topic>DNA, Fungal - isolation & purification</topic><topic>DNA, Intergenic</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>fungi</topic><topic>internal transcribed spacer</topic><topic>internal transcribed spacers</topic><topic>Microbiology</topic><topic>Phylogeny</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Prunus</topic><topic>Prunus dulcis</topic><topic>quantification</topic><topic>quantitative analysis</topic><topic>real-time PCR</topic><topic>Sequence Alignment</topic><topic>soil</topic><topic>Soil Microbiology</topic><topic>Spores, Fungal - growth & development</topic><topic>Spores, Fungal - isolation & purification</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Luo, Y</creatorcontrib><creatorcontrib>Gao, W</creatorcontrib><creatorcontrib>Doster, M</creatorcontrib><creatorcontrib>Michailides, T.J</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>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of applied microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Luo, Y</au><au>Gao, W</au><au>Doster, M</au><au>Michailides, T.J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Quantification of conidial density of Aspergillus flavus and A. parasiticus in soil from almond orchards using real-time PCR</atitle><jtitle>Journal of applied microbiology</jtitle><addtitle>J Appl Microbiol</addtitle><date>2009-05</date><risdate>2009</risdate><volume>106</volume><issue>5</issue><spage>1649</spage><epage>1660</epage><pages>1649-1660</pages><issn>1364-5072</issn><eissn>1365-2672</eissn><abstract>To design the Aspergillus flavus and Aspergillus parasiticus-specific primers and a real-time PCR assay for quantification of the conidial density in soil. Aspergillus flavus and A. parasiticus-specific DNA primers were designed based on internal transcribed spacer sequences to distinguish these two species and from other Aspergillus and other fungal species. A method of pathogen DNA extraction directly from soil samples was developed. Using the designed primers, a real-time PCR assay was developed to quantitatively determine the conidial density of each A. flavus and A. parasiticus in soil, after generating corresponding standard curves. Known conidial densities of each A. flavus or A. parasiticus in soil significantly correlated with those tested with the real-time PCR. This study demonstrated the applicability of the real-time PCR assay in studies of quantifying A. flavus and A. parasiticus in soil as inoculum sources. The A. flacus and A. parasitic-specific primers can be widely used in aflatoxin research. The real-time PCR assay developed in this study provides a potential approach to quantify the plant pathogen density from not only soil but also other sources in relation to aflatoxin contamination from environment, food and feed commodities.</abstract><cop>Oxford, UK</cop><pub>Oxford, UK : Blackwell Publishing Ltd</pub><pmid>19226388</pmid><doi>10.1111/j.1365-2672.2008.04132.x</doi><tpages>12</tpages></addata></record> |
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| source | MEDLINE; Access via Wiley Online Library; Oxford University Press Journals All Titles (1996-Current) |
| subjects | aflatoxin aflatoxins Agriculture Aspergillus - classification Aspergillus - genetics Aspergillus - growth & development Aspergillus - isolation & purification Aspergillus flavus Aspergillus parasiticus Biological and medical sciences DNA Primers - chemistry DNA, Fungal - genetics DNA, Fungal - isolation & purification DNA, Intergenic Fundamental and applied biological sciences. Psychology fungi internal transcribed spacer internal transcribed spacers Microbiology Phylogeny Polymerase Chain Reaction - methods Prunus Prunus dulcis quantification quantitative analysis real-time PCR Sequence Alignment soil Soil Microbiology Spores, Fungal - growth & development Spores, Fungal - isolation & purification |
| title | Quantification of conidial density of Aspergillus flavus and A. parasiticus in soil from almond orchards using real-time PCR |
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