Regeneration and Agrobacterium-mediated transformation studies in carnation (Dianthus caryophyllus L. cv. Turbo)

Abstract

Leaf explants of carnation (Dianthus caryophyllus L. cv. Turbo) were used for the transformation of gene performed by the EHA 105 strain of Agrobacterium tumefaciens harboring the binary vector, pGA482GG. This vector carries the marker genes, neomycin phosphotansferase II (npt II) that determine resistance to kanamycin and -glucoronidase (GUS). Leaf segments of carnation plants were cultured on MS medium containing different combinations and concentrations of hormones. The best shoot
regeneration was observed on MS medium with 1 mg/L BAP+ 0.05 mg/L NAA. Initiation of root formation was performed on MS medium containing 0.5 mg /L IBA. Transformation was done by the EHA 105 strain of A. tumefaciens, after determining the appropriate medium and the plant tissue for the transformation. Plant tissues, selected on MS medium containing kanamycin, were tested by isolating them from transgenic plant tissues and shoots regenerated from these transgenic plants. PCR were
done to indicate transformed GUS gene; 660 bp specific DNA bands of GUS were observed. In this work, an appropriate regeneration system for later studies on carnation and an efficient technique for the transformation of important genes that can resist diseases, using A. tumefaciens were developed.