An Improved Micro Propagation System for Successful Transformation of Cauliflower (Brassica oleracea var. botrytis)

Cauliflower is described as a recalcitrant plant to the genetic transformation process from Agrobacterium. Cauliflower curd meristems were used in a fractionation and sieving protocol to produce microshoots in in vitro culture. This technique produced many thousands of small explants suitable for transfection studies. Explants were successfully transformed following standard transfection with A. tumefaciens carrying a construct with Kanamycin(R), GUS and the Ascorbate Peroxidase (APX) gene. Selection using Kanamycin produced normal plant phenotypic plantlets and these putative transformants were confirmed using GUS staining and PCR of the APX gene sequence. The transfection conditions were optimised during this study and 105 Agrobacterium cells/5 plants/pot successfully produced 1 transformed plant/pot.