A PCR-based assay for detection of Alternaria radicina on carrot seed
A pair of polymerase chain reaction (PCR) primers was developed based upon the sequence of a cloned random amplified polymorphic DNA (RAPD) fragment of Alternaria radicina, and a PCR-based seed assay was developed for the detection of A. radicina from infested carrot seed. The seed assay involved a...
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| Veröffentlicht in: | Plant disease 2001, Vol.85 (1), p.18-23 |
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| container_title | Plant disease |
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| creator | PRYOR, B. M GILBERTSON, R. L |
| description | A pair of polymerase chain reaction (PCR) primers was developed based upon the sequence of a cloned random amplified polymorphic DNA (RAPD) fragment of Alternaria radicina, and a PCR-based seed assay was developed for the detection of A. radicina from infested carrot seed. The seed assay involved a 5-day incubation step, in which seed was maintained under high humidity conditions in order to increase fungal biomass. Seed was then incubated with lysis buffer, extracted with phenol-chloroform, and DNA was recovered using a silica matrix. PCR amplification of the target A. radicina DNA sequence was enhanced by the addition of skim milk to the PCR reaction mixture. With this PCR-based seed assay, A. radicina was detected from carrot seed lots with natural infestation rates as low as 0.3%. In seed lots prepared by mixing known amounts of A. radicina-infested seed with noninfested seed, this assay allowed for the detection of the pathogen from lots with infestation rates as low as 0.1%. |
| doi_str_mv | 10.1094/PDIS.2001.85.1.18 |
| format | Article |
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M ; GILBERTSON, R. L</creator><creatorcontrib>PRYOR, B. M ; GILBERTSON, R. L</creatorcontrib><description>A pair of polymerase chain reaction (PCR) primers was developed based upon the sequence of a cloned random amplified polymorphic DNA (RAPD) fragment of Alternaria radicina, and a PCR-based seed assay was developed for the detection of A. radicina from infested carrot seed. The seed assay involved a 5-day incubation step, in which seed was maintained under high humidity conditions in order to increase fungal biomass. Seed was then incubated with lysis buffer, extracted with phenol-chloroform, and DNA was recovered using a silica matrix. PCR amplification of the target A. radicina DNA sequence was enhanced by the addition of skim milk to the PCR reaction mixture. With this PCR-based seed assay, A. radicina was detected from carrot seed lots with natural infestation rates as low as 0.3%. In seed lots prepared by mixing known amounts of A. radicina-infested seed with noninfested seed, this assay allowed for the detection of the pathogen from lots with infestation rates as low as 0.1%.</description><identifier>ISSN: 0191-2917</identifier><identifier>EISSN: 1943-7692</identifier><identifier>DOI: 10.1094/PDIS.2001.85.1.18</identifier><identifier>PMID: 30832064</identifier><identifier>CODEN: PLDIDE</identifier><language>eng</language><publisher>St. Paul, MN: American Phytopathological Society</publisher><subject>Alternaria radicina ; Biological and medical sciences ; Daucus carota ; Fundamental and applied biological sciences. Psychology ; Fungal plant pathogens ; Generalities. Techniques ; Phytopathology. Animal pests. 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M</creatorcontrib><creatorcontrib>GILBERTSON, R. L</creatorcontrib><title>A PCR-based assay for detection of Alternaria radicina on carrot seed</title><title>Plant disease</title><addtitle>Plant Dis</addtitle><description>A pair of polymerase chain reaction (PCR) primers was developed based upon the sequence of a cloned random amplified polymorphic DNA (RAPD) fragment of Alternaria radicina, and a PCR-based seed assay was developed for the detection of A. radicina from infested carrot seed. The seed assay involved a 5-day incubation step, in which seed was maintained under high humidity conditions in order to increase fungal biomass. Seed was then incubated with lysis buffer, extracted with phenol-chloroform, and DNA was recovered using a silica matrix. PCR amplification of the target A. radicina DNA sequence was enhanced by the addition of skim milk to the PCR reaction mixture. With this PCR-based seed assay, A. radicina was detected from carrot seed lots with natural infestation rates as low as 0.3%. In seed lots prepared by mixing known amounts of A. radicina-infested seed with noninfested seed, this assay allowed for the detection of the pathogen from lots with infestation rates as low as 0.1%.</description><subject>Alternaria radicina</subject><subject>Biological and medical sciences</subject><subject>Daucus carota</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fungal plant pathogens</subject><subject>Generalities. Techniques</subject><subject>Phytopathology. Animal pests. Plant and forest protection</subject><subject>seeds</subject><issn>0191-2917</issn><issn>1943-7692</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><recordid>eNp9kE1LxDAQhoMoun78AC8SEMRL60yaNslxWT9hwcWPc5hNE6h0W026h_33dnH16GkG5nlfhoexc4Qcwcibxe3Tay4AMNdljjnqPTZBI4tMVUbsswmgwUwYVEfsOKUPAJCy0ofsqABdCKjkhN1N-WL2ki0p-ZpTSrThoY-89oN3Q9N3vA982g4-dhQb4pHqxjUd8fHiKMZ-4Mn7-pQdBGqTP9vNE_Z-f_c2e8zmzw9Ps-k8cxJwyCgoVZqlJyQSVAKGytSqWFbGQZAKA0lUevwSal1WOngTCgxeFugEjXtxwq5_ej9j_7X2abCrJjnfttT5fp2sQK0FSAFqRK_-RVGpShphRhB_QBf7lKIP9jM2K4obi2C3mu1Ws91qtrq0aFGPmYtd-Xq58vVf4tfrCFzuAEqO2hCpc0364wwoZUTxDZrsgxU</recordid><startdate>2001</startdate><enddate>2001</enddate><creator>PRYOR, B. M</creator><creator>GILBERTSON, R. 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Plant and forest protection</topic><topic>seeds</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>PRYOR, B. M</creatorcontrib><creatorcontrib>GILBERTSON, R. L</creatorcontrib><collection>Pascal-Francis</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>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Plant disease</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>PRYOR, B. M</au><au>GILBERTSON, R. L</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A PCR-based assay for detection of Alternaria radicina on carrot seed</atitle><jtitle>Plant disease</jtitle><addtitle>Plant Dis</addtitle><date>2001</date><risdate>2001</risdate><volume>85</volume><issue>1</issue><spage>18</spage><epage>23</epage><pages>18-23</pages><issn>0191-2917</issn><eissn>1943-7692</eissn><coden>PLDIDE</coden><abstract>A pair of polymerase chain reaction (PCR) primers was developed based upon the sequence of a cloned random amplified polymorphic DNA (RAPD) fragment of Alternaria radicina, and a PCR-based seed assay was developed for the detection of A. radicina from infested carrot seed. The seed assay involved a 5-day incubation step, in which seed was maintained under high humidity conditions in order to increase fungal biomass. Seed was then incubated with lysis buffer, extracted with phenol-chloroform, and DNA was recovered using a silica matrix. PCR amplification of the target A. radicina DNA sequence was enhanced by the addition of skim milk to the PCR reaction mixture. With this PCR-based seed assay, A. radicina was detected from carrot seed lots with natural infestation rates as low as 0.3%. In seed lots prepared by mixing known amounts of A. radicina-infested seed with noninfested seed, this assay allowed for the detection of the pathogen from lots with infestation rates as low as 0.1%.</abstract><cop>St. Paul, MN</cop><pub>American Phytopathological Society</pub><pmid>30832064</pmid><doi>10.1094/PDIS.2001.85.1.18</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 0191-2917 |
| ispartof | Plant disease, 2001, Vol.85 (1), p.18-23 |
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| language | eng |
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| source | Alma/SFX Local Collection; EZB Electronic Journals Library; American Phytopathological Society Journal Back Issues |
| subjects | Alternaria radicina Biological and medical sciences Daucus carota Fundamental and applied biological sciences. Psychology Fungal plant pathogens Generalities. Techniques Phytopathology. Animal pests. Plant and forest protection seeds |
| title | A PCR-based assay for detection of Alternaria radicina on carrot seed |
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