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ISHS Acta Horticulturae 560: IV International Symposium on In Vitro Culture and Horticultural Breeding

STABLE TRANSFORMATION OF PIONSETTIA VIA ELECTROPHORESIS

Authors:   N.I. Vik, A.K. Hvoslef-Eide, H. Gjerde, K. Bakke
Keywords:   Euphorbia pulcherrima, genetic transformation, GUS, in vivo electroporesis
DOI:   10.17660/ActaHortic.2001.560.12
Abstract:
As a result of our interest in poinsettia breeding, we started out with poinsettia transformations and tested results of three different gene transfer methods: with Agrobacterium, particle gun delivery and in vivo electrophoresis. The latter seems both interesting and promising as a method of transforming intact poinsettia meristems. Young poinsettia plants were topped to promote growth of axillary shoots and meristems of these shoots were blotted. Plasmid pJE101, which includes the GUS gene, was used for the transformations. 1mg DNA/ml was cast in 1 ml pipette-tips in a 0,5% agarose solution and placed on top of an exposed meristem. The negative electrode was placed in the agarose solution and the positive electrode in the soil at the base of the stem producing a current through the meristem. Most meristems were subjected to a current ~0,5 mA for 10 minutes. GUS analysis was performed 8-9 weeks after electrophoresis and 9 of 34 shoots tested appeared to be positive. One of the positive GUS plants was further tested by Southern analysis one year after electrophoresis and showed clear bands for the predicted transformed material. We have continued our work with the method of electrophoresis in poinsettia and have been transforming plants with genes of interest, such as the growth regulating rolC gene.

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