Quantitative Molecular Detection of Xanthomonas hortorum pv. carotae in Carrot Seed Before and After Hot-Water Treatment
Molecular assays to detect and quantify DNA from viable cells of the seedborne pathogen Xanthomonas hortorum pv. carotae in carrot seed were developed and evaluated for use on nontreated and hot-water-treated seed lots. Both a TaqMan real-time polymerase chain reaction (PCR) assay and a loop-mediate...
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| Veröffentlicht in: | Plant disease 2013-12, Vol.97 (12), p.1585-1592 |
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| creator | TEMPLE, Todd N DU TOIT, Lindsey J DERIE, Michael L JOHNSON, Kenneth B |
| description | Molecular assays to detect and quantify DNA from viable cells of the seedborne pathogen Xanthomonas hortorum pv. carotae in carrot seed were developed and evaluated for use on nontreated and hot-water-treated seed lots. Both a TaqMan real-time polymerase chain reaction (PCR) assay and a loop-mediated isothermal amplification (LAMP) dilution endpoint assay detected and quantified DNA from viable pathogen cells after treatment of carrot seed washes with the live-dead discriminating dye propidium monoazide (PMA). The detection limits of the assays were approximately 10
CFU for pure cultures of X. hortorum pv. carotae, and 10
to 10
CFU/g seed from naturally infested carrot seed lots. X. hortorum pv. carotae in and on carrot seed was killed by soaking the seed in hot water (52°C for 25 min), and a subsequent PMA treatment of these hot-water-treated seed washes suppressed detection of the pathogen with both the real-time PCR and LAMP assays. For 36 commercial seed lots treated with PMA but not hot water, regression of colony counts of X. hortorum pv. carotae measured by dilution plating on a semiselective agar medium versus estimates of pathogen CFU determined by the molecular assays resulted in significant (P ≤ 0.05) linear relationships (R
= 0.68 for the real-time PCR assay and 0.79 for the LAMP assay). The molecular assays provided quantitative estimates of X. hortorum pv. carotae infestations in carrot seed lots in |
| doi_str_mv | 10.1094/PDIS-03-13-0262-RE |
| format | Article |
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CFU for pure cultures of X. hortorum pv. carotae, and 10
to 10
CFU/g seed from naturally infested carrot seed lots. X. hortorum pv. carotae in and on carrot seed was killed by soaking the seed in hot water (52°C for 25 min), and a subsequent PMA treatment of these hot-water-treated seed washes suppressed detection of the pathogen with both the real-time PCR and LAMP assays. For 36 commercial seed lots treated with PMA but not hot water, regression of colony counts of X. hortorum pv. carotae measured by dilution plating on a semiselective agar medium versus estimates of pathogen CFU determined by the molecular assays resulted in significant (P ≤ 0.05) linear relationships (R
= 0.68 for the real-time PCR assay and 0.79 for the LAMP assay). The molecular assays provided quantitative estimates of X. hortorum pv. carotae infestations in carrot seed lots in <24 h, which is a significant improvement over the 7 to 14 days required to obtain results from the traditional dilution-plating assay.</description><identifier>ISSN: 0191-2917</identifier><identifier>EISSN: 1943-7692</identifier><identifier>DOI: 10.1094/PDIS-03-13-0262-RE</identifier><identifier>PMID: 30716831</identifier><identifier>CODEN: PLDIDE</identifier><language>eng</language><publisher>St. Paul, MN: American Phytopathological Society</publisher><subject>Biological and medical sciences ; Daucus ; Fundamental and applied biological sciences. Psychology ; Phytopathology. Animal pests. Plant and forest protection ; Xanthomonas</subject><ispartof>Plant disease, 2013-12, Vol.97 (12), p.1585-1592</ispartof><rights>2015 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c366t-f44608e3d7d3242d758825d8d27e4e36d1bda30a3b6c0673cf4b394d0fa51a63</citedby><cites>FETCH-LOGICAL-c366t-f44608e3d7d3242d758825d8d27e4e36d1bda30a3b6c0673cf4b394d0fa51a63</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,3724,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=27960316$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30716831$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>TEMPLE, Todd N</creatorcontrib><creatorcontrib>DU TOIT, Lindsey J</creatorcontrib><creatorcontrib>DERIE, Michael L</creatorcontrib><creatorcontrib>JOHNSON, Kenneth B</creatorcontrib><title>Quantitative Molecular Detection of Xanthomonas hortorum pv. carotae in Carrot Seed Before and After Hot-Water Treatment</title><title>Plant disease</title><addtitle>Plant Dis</addtitle><description>Molecular assays to detect and quantify DNA from viable cells of the seedborne pathogen Xanthomonas hortorum pv. carotae in carrot seed were developed and evaluated for use on nontreated and hot-water-treated seed lots. Both a TaqMan real-time polymerase chain reaction (PCR) assay and a loop-mediated isothermal amplification (LAMP) dilution endpoint assay detected and quantified DNA from viable pathogen cells after treatment of carrot seed washes with the live-dead discriminating dye propidium monoazide (PMA). The detection limits of the assays were approximately 10
CFU for pure cultures of X. hortorum pv. carotae, and 10
to 10
CFU/g seed from naturally infested carrot seed lots. X. hortorum pv. carotae in and on carrot seed was killed by soaking the seed in hot water (52°C for 25 min), and a subsequent PMA treatment of these hot-water-treated seed washes suppressed detection of the pathogen with both the real-time PCR and LAMP assays. For 36 commercial seed lots treated with PMA but not hot water, regression of colony counts of X. hortorum pv. carotae measured by dilution plating on a semiselective agar medium versus estimates of pathogen CFU determined by the molecular assays resulted in significant (P ≤ 0.05) linear relationships (R
= 0.68 for the real-time PCR assay and 0.79 for the LAMP assay). The molecular assays provided quantitative estimates of X. hortorum pv. carotae infestations in carrot seed lots in <24 h, which is a significant improvement over the 7 to 14 days required to obtain results from the traditional dilution-plating assay.</description><subject>Biological and medical sciences</subject><subject>Daucus</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Phytopathology. Animal pests. 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Psychology</topic><topic>Phytopathology. Animal pests. Plant and forest protection</topic><topic>Xanthomonas</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>TEMPLE, Todd N</creatorcontrib><creatorcontrib>DU TOIT, Lindsey J</creatorcontrib><creatorcontrib>DERIE, Michael L</creatorcontrib><creatorcontrib>JOHNSON, Kenneth B</creatorcontrib><collection>Pascal-Francis</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aqualine</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Water Resources Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Engineering Research Database</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 3: Aquatic Pollution & Environmental Quality</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</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>TEMPLE, Todd N</au><au>DU TOIT, Lindsey J</au><au>DERIE, Michael L</au><au>JOHNSON, Kenneth B</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Quantitative Molecular Detection of Xanthomonas hortorum pv. carotae in Carrot Seed Before and After Hot-Water Treatment</atitle><jtitle>Plant disease</jtitle><addtitle>Plant Dis</addtitle><date>2013-12-01</date><risdate>2013</risdate><volume>97</volume><issue>12</issue><spage>1585</spage><epage>1592</epage><pages>1585-1592</pages><issn>0191-2917</issn><eissn>1943-7692</eissn><coden>PLDIDE</coden><abstract>Molecular assays to detect and quantify DNA from viable cells of the seedborne pathogen Xanthomonas hortorum pv. carotae in carrot seed were developed and evaluated for use on nontreated and hot-water-treated seed lots. Both a TaqMan real-time polymerase chain reaction (PCR) assay and a loop-mediated isothermal amplification (LAMP) dilution endpoint assay detected and quantified DNA from viable pathogen cells after treatment of carrot seed washes with the live-dead discriminating dye propidium monoazide (PMA). The detection limits of the assays were approximately 10
CFU for pure cultures of X. hortorum pv. carotae, and 10
to 10
CFU/g seed from naturally infested carrot seed lots. X. hortorum pv. carotae in and on carrot seed was killed by soaking the seed in hot water (52°C for 25 min), and a subsequent PMA treatment of these hot-water-treated seed washes suppressed detection of the pathogen with both the real-time PCR and LAMP assays. For 36 commercial seed lots treated with PMA but not hot water, regression of colony counts of X. hortorum pv. carotae measured by dilution plating on a semiselective agar medium versus estimates of pathogen CFU determined by the molecular assays resulted in significant (P ≤ 0.05) linear relationships (R
= 0.68 for the real-time PCR assay and 0.79 for the LAMP assay). The molecular assays provided quantitative estimates of X. hortorum pv. carotae infestations in carrot seed lots in <24 h, which is a significant improvement over the 7 to 14 days required to obtain results from the traditional dilution-plating assay.</abstract><cop>St. Paul, MN</cop><pub>American Phytopathological Society</pub><pmid>30716831</pmid><doi>10.1094/PDIS-03-13-0262-RE</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| source | Free E-Journal (出版社公開部分のみ）; Alma/SFX Local Collection; American Phytopathological Society Journal Back Issues |
| subjects | Biological and medical sciences Daucus Fundamental and applied biological sciences. Psychology Phytopathology. Animal pests. Plant and forest protection Xanthomonas |
| title | Quantitative Molecular Detection of Xanthomonas hortorum pv. carotae in Carrot Seed Before and After Hot-Water Treatment |
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