The Potential for Green Fluorescent Protein as a Screening Tool in the Production of Haploid Potato Plants

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dc.contributor.authorPalumbo, Roseen
dc.contributor.committeechairVeilleux, Richard E.en
dc.contributor.committeememberShulaev, Vladimiren
dc.contributor.committeememberWelbaum, Gregory E.en
dc.contributor.departmentHorticultureen
dc.date.accessioned2011-08-06T14:44:16Zen
dc.date.adate2003-12-31en
dc.date.available2011-08-06T14:44:16Zen
dc.date.issued2003-12-17en
dc.date.rdate2004-12-31en
dc.date.sdate2003-12-19en
dc.description.abstractA hybrid between a highly regenerative diploid clone (BARD 1-3) of Solanum phureja and haploid inducer IVP 101 was transformed with Agrobacterium tumefaciens strain 4404 containing plasmid pHB2892 with genes for green florescent protein (GFP) and kanamycin resistance. Hemizygous primary transformants (To) were produced from three leaf discs: 17 diploid plants from one leaf disc, three and nine tetraploids from the other two leaf discs. GFP expression was observed qualitatively under fluorescence microscopes and quantitatively with a GFP meter. Anther culture of tetraploids produced 29 plants, none with high levels of GFP. Segregation ratios for tetraploid T1 seedlings fit models for single duplex insertions (35 transgenic: 1 non) or double simplex insertions (15 transgenic: 1 non). Diploid T1 seedlings segregated for deleterious traits: dwarfed size and curled leaves, as well as the GFP transgene. Similar segregation patterns in diploid families implied that all diploids may have been from the same transformation event. The cumulative segregation showed the dwarfed and curled plants fit a single recessive gene ratio (3 normal: 1 mutant), and GFP fit a double-copy insertion ratio (15 transgenic: 1 non). There was substantial GFP silencing evidenced by the loss of expression in plants that had originally been selected for high GFP. However, six selections were found to be free of deleterious traits, consistently high expressers of GFP, and producers of stainable pollen with less 2n than IVP 101.en
dc.description.degreeMaster of Scienceen
dc.format.mediumETDen
dc.identifier.otheretd-12192003-162452en
dc.identifier.sourceurlhttp://scholar.lib.vt.edu/theses/available/etd-12192003-162452en
dc.identifier.urihttp://hdl.handle.net/10919/9651en
dc.publisherVirginia Techen
dc.relation.haspartRosiesThesis.pdfen
dc.rightsIn Copyrighten
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.subjecttransgeneic plantsen
dc.subjectSolanum tuberosumen
dc.subjectSolanum phurejaen
dc.subjecthaploid-inducing pollinatoren
dc.subjectGFPen
dc.subjectgenetic transformationen
dc.titleThe Potential for Green Fluorescent Protein as a Screening Tool in the Production of Haploid Potato Plantsen
dc.typeThesisen
thesis.degree.disciplineHorticultureen
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen
thesis.degree.levelmastersen
thesis.degree.nameMaster of Scienceen

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