Authors: Isabel MG Padilla Agnieszka Golis Adele Gentile Carmine Damiano Ralph Scorza
Publish Date: 2006/01/25
Volume: 84, Issue: 3, Pages: 309-314
Abstract
To determine the optimum conditions for Agrobacteriummediated gene transfer peach explants including cotyledons embryonic axes and hypocotyl slices from nongerminated seeds and epicotyl internode slices from germinating seeds were exposed to Agrobacteriummediated transformation treatments The GUS uidA marker gene was tested using two different A tumefaciens strains three plasmids and four promoters CaMV35s Aocs3AmasPmas “superpromoter” masCaMV35s and CAB GFP was tested with six A␣tumefaciens strains one plasmid pLC101 and the doubleCaMV35s dCaMV35s promoter The CaMV35s promoter produced more GUS expression than the CAB promoter A tumefaciens strains EHA105 and LBA4404 harboring the same plasmid pBIN19 differed in their effects on GUS expression suggesting an interaction between A tumefaciens strain and plasmid A combination of A tumefaciens EHA105 plasmid pBIN19 and the CaMV35s promoter produced the highest rates of transformation in peach epicotyl internodes 568 cotyledons 527 leaves 20 and embryonic axes 467 as evaluated by the percentage of explants expressing GUS 14 days after cocultivation GFP expression under the control of the dCaMV35s promoter was highest for internode explants but only reached levels of 18–19 When GFPcontaining plasmid pCL101 was combined with each of five A tumefaciens strains the highest levels of transformation were 20–21 internode and cotyledons respectively When nine peach genotypes were cocultivated with A tumefaciens strain EHA105 and GFPcontaining plasmid pCL101 the highest levels of transformation were 26–28 cotyledons and internodes respectively While GFP represents a potentially useful transformation marker that allows the nondestructive evaluation of transformation rates of GFP transformation under the conditions of this study were low It will be necessary to optimize expression of this marker gene in peach
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