title: Transposon based activation tagging in diploid strawberry and monoploid derivatives of potato.
abstract: Diploid strawberry and potato transformed with a transposon tagging construct exhibited either global (strawberry) or local transposition (potato). An activation tagged, compact-sized strawberry mutant overexpressed the gene adjacent to Ds. As major fruit and vegetable crops, respectively, strawberry and potato are among the first horticultural crops with draft genome sequences. To study gene function, we examined transposon-tagged mutant strategies in model populations for both species, Fragaria vesca and Solanum tuberosum Group Phureja, using the same Activation/Dissociation (Ac/Ds) construct. Early somatic transposition during tissue culture occurred at a frequency of 18.5% in strawberry but not in potato transformants. Green fluorescent protein under a monocot promoter was a more reliable selectable marker in strawberry compared to potato. BASTA (gluphosinate herbicide) resistance served as an effective selectable marker for both species (80 and 85% reliable in strawberry and potato, respectively), although the effective concentration differed (0.5% for strawberry and 0.03% for potato). Transposons preferentially reinserted within genes (exons and introns) in both species. Real-time quantitative PCR revealed enhanced gene expression (670 and 298-fold expression compared to wild type in petiole and leaf tissue, respectively) for an activation tagged strawberry mutant with Ds inserted about 0.6 kb upstream from a gene coding for an epidermis-specific secreted glycoprotein EP1. Our data also suggested that endopolyploid (diploid) cells occurring in leaf explants of monoploid potato were the favored targets of T-DNA integration during transformation. Mutants obtained in these studies provide a useful resource for future genetic studies.
authors.affiliation: Department of Horticulture, Virginia Polytechnic Institute and State University, Blacksburg, VA, 24061, USA.
authors.forename: Nan
authors.initials: N
authors.lastname: Lu
authors.forename: Jared D
authors.initials: JD
authors.lastname: Carter
authors.forename: Tatiana
authors.initials: T
authors.lastname: Boluarte Medina
authors.forename: Sarah H
authors.initials: SH
authors.lastname: Holt
authors.forename: Norma Constanza
authors.initials: NC
authors.lastname: Manrique-Carpintero
authors.forename: Kendall T
authors.initials: KT
authors.lastname: Upham
authors.forename: Andy
authors.initials: A
authors.lastname: Pereira
authors.forename: Vladimir
authors.initials: V
authors.lastname: Shulaev
authors.forename: Richard E
authors.initials: RE
authors.lastname: Veilleux
mesh_terms: D060054:Agrobacterium / Q000235:genetics; D001483:Base Sequence; D018556:Crops, Agricultural / Q000235:genetics; D004251:DNA Transposable Elements; D004171:Diploidy; D031985:Fragaria / Q000235:genetics*; D018525:Germination; D008969:Molecular Sequence Data; D009154:Mutation; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction / Q000379:methods; D011198:Solanum tuberosum / Q000235:genetics*; D014170:Transformation, Genetic
chemical_list: D004251:DNA Transposable Elements
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