180 GENETIC TRANSFORMATION OF RED RASPBERRY (RUBUS IDAEUS L.) WITH A GENE TO CONTROL ETHYLENE BIOSYNTHESIS

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  • 1 Agritope Inc. 8505 SW Creekside Place, Beaverton, Oregon 97005

We have developed efficient plant rageneration and transformation systems for red raspberry (Rubus idaeus L.). We have successfully introduced a gene for controlling biosynthesis of ethylene into raspberry for the first time. Leaf and petiole segments were co-cultivated with disarmed Agrobacterium strains EHA 101 or 105 containing plasmids pAG5420, pAG 1452 or pAG1552. The plasmids encoded gene sequences for S-adenosylmethionine hydrolase (SAM ase) driven by the fruit specific or wound and fruit specific tomato SE8 or E4 promoters. SAM ase catalyses the conversion of S-adenosylmethionine (SAM) to methylthioadenosine (MTA) and homoserine which can reenter the methionine recycling pathway. SAM is therefore not available for the synthesis of 1-am inocyclopropane carboxylic acid (ACC), the metabolic precursor for ethylene biosynthesis. Initial shoot regenerants were mostly chimeras containing transformed and non-transformed cells. Solid clones of pure transgenics were developed by repeated culture of leaf, petiole and nodal explants of primary regenerants on higher stringency selection medium. Transformants were screened on medium with kanamycin, geneticin or hygromycin depending on the selection marker gene NPTII or hpt. Genomic integration of transgenes were confirmed by Southern hybridization. Transgenic plants of cultivars Canby, Meeker and Chilliwack have been transplanted to the greenhouse for fruit set and further evaluation of transgenic traits.

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