Protoplast isolation prior to flow cytometry reveals clear patterns of endoreduplication in potato tubers, related species, and some starchy root crops

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dc.contributor.authorLaimbeer, F. Parker E.en
dc.contributor.authorHolt, Sarah H.en
dc.contributor.authorMakris, Melissaen
dc.contributor.authorHardigan, Michael Alanen
dc.contributor.authorBuell, C. Robinen
dc.contributor.authorVeilleux, Richard E.en
dc.contributor.departmentBiomedical Sciences and Pathobiologyen
dc.contributor.departmentSchool of Plant and Environmental Sciencesen
dc.contributor.departmentVirginia-Maryland College of Veterinary Medicineen
dc.date.accessioned2018-01-17T18:29:41Zen
dc.date.available2018-01-17T18:29:41Zen
dc.date.issued2017-04-14en
dc.description.abstractBackground: Endoreduplication, the process of DNA replication in the absence of cell division, is associated with specialized cellular function and increased cell size. Genes controlling endoreduplication in tomato fruit have been shown to affect mature fruit size. An efficient method of estimating endoreduplication is required to study its role in plant organ development. Flow cytometry is often utilized to evaluate endoreduplication, yet some tissues and species, among them the tubers of Solanum tuberosum, remain intractable to routine tissue preparation for flow cytometry. We aimed to develop a method through the use of protoplast extraction preceding flow cytometry, specifically for the assessment of endoreduplication in potato tubers. Results: We present a method for appraising endoreduplication in potato (Solanum tuberosum) tuber tissues. We evaluated this method and observed consistent differences between pith and cortex of tubers and between different cultivars, but no apparent relationship with whole tuber size. Furthermore, we were able to observe distinct patterns of endoreduplication in 16 of 20 wild potato relatives, with mean endoreduplication index (EI) ranging from 0.94 to 2.62 endocycles per cell. The protocol was also applied to a panel of starchy root crop species and, while only two of five yielded reliable flow histograms, the two (sweet potato and turnip) exhibited substantially lower EIs than wild and cultivated potato accessions. Conclusions: The protocol reported herein has proven effective on tubers of a variety of potato cultivars and related species, as well as storage roots of other starchy crops. This method provides an important tool for the study of potato morphology and development while revealing natural variation for endoreduplication which may have agricultural relevance.en
dc.description.versionPublished versionen
dc.format.extent? - ? (10) page(s)en
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.118/s13007-017-0177-3en
dc.identifier.issn1746-4811en
dc.identifier.issue1en
dc.identifier.urihttp://hdl.handle.net/10919/81829en
dc.identifier.volume13en
dc.language.isoenen
dc.publisherBiomed Centralen
dc.relation.urihttp://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000399462700002&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1en
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectBiochemical Research Methodsen
dc.subjectPlant Sciencesen
dc.subjectBiochemistry & Molecular Biologyen
dc.subjectSolanaceaeen
dc.subjectEndopolyploidizationen
dc.subjectSolanum tuberosumen
dc.subjectKaryoplasmic ratioen
dc.subjectCELL-DIVISIONen
dc.subjectELEVATED CO2en
dc.subjectNUCLEAR-DNAen
dc.subjectPLANTSen
dc.subjectGROWTHen
dc.subjectARABIDOPSISen
dc.subjectTISSUESen
dc.subjectCYCLEen
dc.titleProtoplast isolation prior to flow cytometry reveals clear patterns of endoreduplication in potato tubers, related species, and some starchy root cropsen
dc.title.serialPlant Methodsen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
pubs.organisational-group/Virginia Techen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciencesen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/CALS T&R Facultyen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/Horticultureen
pubs.organisational-group/Virginia Tech/All T&R Facultyen
pubs.organisational-group/Virginia Tech/University Research Institutesen
pubs.organisational-group/Virginia Tech/University Research Institutes/Fralin Life Sciencesen
pubs.organisational-group/Virginia Tech/University Research Institutes/Fralin Life Sciences/Fralin Affiliated Facultyen

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