<i>Agrobacterium</i>-mediated transformation and regeneration of transgenic 'Chancellor' wine grape plants expressing the <i>tfd</i>A gene

Authors

  • R. M. S. Mulwa
  • M. A. Norton
  • S. K. Farrand
  • R. M. Skirvin

DOI:

https://doi.org/10.5073/vitis.2007.46.110-115

Keywords:

‘Chancellor’ grapes, somatic embryogenesis, tfdA gene, genetic transformation, 2, 4-D = 2, 4-dichlorophenoxyacetic acid, herbicide resistance

Abstract

The establishment of grape vineyards in the Midwestern USA is greatly hampered by the use of the herbicide 2,4-D in grain crop fields. The overall goal of this study was to transform 'Chancellor' with the plant expressible bacterial tfdA gene to make it tolerant to 2,4-D. Embryogenic callus was infected and cocultivated with an Agrobacterium construct (LBA4404: pAL4404::tfdA); transformed callus was selected with kanamycin; kanamycin resistant callus was bulked and transformed cell lines identified by PCR. Three PCRpositive embryogenic callus lines were used to regenerate transgenic plants. Analysis of the plant lines for the presence of the tfdA gene by PCR and southern hybridization confirmed its stable integration in their genomes. Transgenic 'Chancellor' grape plants regenerated from these calli proved to be resistant to up to 10 kg·ha-1 of a commercial ester-formulation of 2,4-D, indicating positive expression of the tfdA gene and affording them protection from 2,4-D injury.


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Published

2015-04-14

Issue

Vol. 46 No. 3 (2007): Vitis

Section

Article

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