Show simple item record

dc.contributor.authorNie, X.
dc.contributor.authorDickison, V.L.
dc.contributor.authorBrooks, S.
dc.contributor.authorNie, B.
dc.contributor.authorSingh, M.
dc.contributor.authorKoeyer, D.L. de
dc.contributor.authorMurphy, A.M.
dc.date.accessioned2019-12-04T11:11:19Z
dc.date.available2019-12-04T11:11:19Z
dc.date.issued2017
dc.identifier.citationNie, 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.
dc.identifier.issn0191-2917
dc.identifier.urihttps://hdl.handle.net/20.500.12478/2402
dc.description.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.
dc.description.sponsorshipAgriculture and Agri-Food Canada
dc.format.extent1-36
dc.language.isoen
dc.subjectGene
dc.subjectPotato
dc.subjectDna
dc.subjectGenotypes
dc.subjectPotato Virus
dc.subjectHigh Resolution
dc.subjectResistance
dc.subjectCultivars
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 potato
dc.typeJournal Article
dc.description.versionPeer Review
cg.contributor.crpRoots, Tubers and Bananas
cg.contributor.affiliationAgriculture and Agri-Food Canada
cg.contributor.affiliationUniversity of Victoria
cg.contributor.affiliationHuazhong Agricultural University
cg.contributor.affiliationAgricultural Certification Services, Canada
cg.contributor.affiliationInternational Institute of Tropical Agriculture
cg.isijournalISI Journal
cg.authorship.typesCGIAR and advanced research institute
cg.iitasubjectGenetic Improvement
cg.iitasubjectPlant Diseases
cg.iitasubjectPlant Genetic Resources
cg.journalPlant Disease
cg.howpublishedFormally Published
cg.accessibilitystatusLimited Access
local.dspaceid92369
cg.targetaudienceScientists
cg.identifier.doihttp://dx.doi.org/10.1094/pdis-07-17-0968-re


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record