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Co-transformation of Grapevine Somatic Embryos to Produce Transgenic Plants Free of Marker Genes

 Last Update: 2021-03-14
 Source: Internet
 Author: User

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A cotransformation system using somatic embryos was developed to produce grapevines free of selectable marker genes. This was achieved by transforming
Vitis vinifera
L. “Thompson Seedless” somatic embryos with a mixture of two
Agrobacterium
strains. The first strain contained a binary plasmid with an
egfp
gene of interest between the T-
DNA
borders. The second strain harbored the neomycin phosphotransferase (
npt
II) gene for positive selection and the cytosine deaminase (
codA
) gene for negative selection, linked together by a bidirectional dual promoter complex. Our technique included a short positive selection phase of cotransformed somatic embryos on liquid medium containing 100 mg/L kanamycin before subjecting cultures to prolonged negative selection on medium containing 250 mg/L 5-fluorocytosine.

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