Genotype Independent Pollen Mediated
Transformation in Wheat- A new Approach? (5790)
Authors:
M. Kumar* - University of Idaho J.L. Hansen - University of Idaho R.S. Zemetra - University of Idaho
Abstract:
Genetic transformation of wheat (Triticum aestivum) has the potential to facilitate the introduction of genes for rapid response to new biotic and abiotic stresses. One problem has been the genotype dependent nature of wheat transformation. Pollen mediated transformation where electroporation is used to deliver foreign DNA into mature pollen prior to pollination offers the potential for a genotype independent transformation system. The viability of pollen before and after electroporation was evaluated for four winter wheat cultivars; Brundage, Hubbard, Daws, and Madsen.
There was no interaction between cultivar and treatment indicating wheat pollen response to electroporation was genotype independent. A two-dye optimization protocol was used to optimize electroporation conditions. 1.03 kV/cm was found to be the optimum voltage for wheat pollen electroporation. Wheat pollen was
electroporated with the plasmid pDPPG165 carrying the bar gene for
phosphinothricin resistance. A total of 21 seed were obtained from pollen-mediated transformation of which 20 germinated. Of the germinated seed 50% were found to carry the bar gene based on PCR analysis. Forty percent of the seedlings positive for the bar gene were positive for gene expression based on a leaf swipe test for herbicide resistance. This is the first report of successful transformation of wheat by pollen-mediated transformation and may potentially provide a
genotype-independent form of transformation of wheat.
Speaker Information: Manish Kumar, University of Idaho, 115.S.lilley Apt # 201, Moscow, ID 83843; Phone: 208-882-0306; E-mail: [email protected]
Session Information: Monday, November 1, 2004, 2:00 PM-4:00 PM Presentation Start: 2:00 PM (Poster Board Number: 1470)
Keywords: Wheat; Pollen; Electroporation; Genotype independent transformation
