High resolution DNA melting assays for detection of Rx1 and Rx2 for high-throughput marker assisted selection for extreme resistance to Potato virus X in tetraploid potato

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cg.authorship.typesCGIAR and advanced research instituteen_US
cg.contributor.affiliationAgriculture and Agri-Food Canadaen_US
cg.contributor.affiliationUniversity of Victoriaen_US
cg.contributor.affiliationHuazhong Agricultural Universityen_US
cg.contributor.affiliationAgricultural Certification Services, Canadaen_US
cg.contributor.affiliationInternational Institute of Tropical Agricultureen_US
cg.contributor.crpRoots, Tubers and Bananasen_US
cg.contributor.donorAgriculture and Agri-Food Canadaen_US
cg.howPublishedFormally Publisheden_US
cg.identifier.doihttps://doi.org/10.1094/pdis-07-17-0968-reen_US
cg.isijournalISI Journalen_US
cg.issn0191-2917en_US
cg.issue2en_US
cg.journalPlant Diseaseen_US
cg.reviewStatusPeer Reviewen_US
cg.subject.iitaGENETIC IMPROVEMENTen_US
cg.subject.iitaPLANT DISEASESen_US
cg.subject.iitaPLANT GENETIC RESOURCESen_US
cg.volume102en_US
dc.contributor.authorNie, X.en_US
dc.contributor.authorDickison, V.L.en_US
dc.contributor.authorBrooks, S.en_US
dc.contributor.authorNie, B.en_US
dc.contributor.authorSingh, M.en_US
dc.contributor.authorKoeyer, D.L. deen_US
dc.contributor.authorMurphy, A.M.en_US
dc.date.accessioned2018-01-08T12:49:59Zen_US
dc.date.available2018-01-08T12:49:59Zen_US
dc.identifier.urihttps://hdl.handle.net/10568/89939en_US
dc.titleHigh resolution DNA melting assays for detection of Rx1 and Rx2 for high-throughput marker assisted selection for extreme resistance to Potato virus X in tetraploid potatoen_US
dcterms.abstractAssessment of the existing PCR-gel electrophoresis-based methods for detection of Rx1 and Rx2, the genes that independently control extreme resistance (ER) to Potato virus X (PVX), indicated that the 5Rx1F/5Rx1R primer pair led to reliable detection of Rx1 whereas the 106Rx2F/106Rx2R primer pair detected Rx2 despite some non-specific reactions in potato clones/cultivars without Rx2. However, the methodology is time consuming and does not differentiate the absence of Rx1/Rx2 from a failed PCR reaction. A newly designed primer pair that targets Rx1 and Rx2 as well as rx1 and rx2 produced an amplicon for all alleles. When the primer pair was combined with 5Rx1F/5Rx1R, respective amplicons were produced, although they were not distinguishable by regular agarose gel electrophoresis. When subjected to a high resolution DNA melting (HRM) assay, two distinct melting profiles for Rx1 and rx1, respectively, were detected. Triplex PCR-gel electrophoresis and -HRM assay for detection of Rx1, Rx2 and rx1/rx2 were also performed. The efficacy of the HRM assays were validated in potato cultivars/clones with known phenotypes, indicating its potential for high-throughput selection of potato clones/cultivars carrying Rx1 or Rx2. Duplex PCR-HRM assays of over 600 progeny from 12 crosses involving various parents correctly detected the presence or absence of Rx1 in each progeny, allowing accurate prediction of the phenotype. Progeny that tested positive for Rx1 by HRM exhibited ER to PVX whereas progeny that tested negative for Rx1 were susceptible to PVX infection. The genotype of each parent and the possible presence of Nx in two Rx1-possessing parents are also discussed.en_US
dcterms.accessRightsOpen Accessen_US
dcterms.audienceScientistsen_US
dcterms.bibliographicCitationNie, X., Dickison, V.L., Brooks, S., Nie, B., Singh, M., De Koeyer, D.L. & Murphy, A.M. (2017). High resolution DNA melting assays for detection of Rx1 and Rx2 for high-throughput marker-assisted selection for extreme resistance to Potato virus X in tetraploid potato. Plant Disease, 1-36.en_US
dcterms.extentp. 382-390en_US
dcterms.issued2018-02-01en_US
dcterms.languageenen_US
dcterms.publisherScientific Societiesen_US
dcterms.subjectgeneen_US
dcterms.subjectpotatoen_US
dcterms.subjectdnaen_US
dcterms.subjectgenotypesen_US
dcterms.subjectpotato virusen_US
dcterms.subjecthigh resolutionen_US
dcterms.subjectresistanceen_US
dcterms.subjectcultivarsen_US
dcterms.typeJournal Articleen_US

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