Protoplast Culture and Fusion between Brassica Carinata and Brassica Napus

In this study, attempts were made to select in vitro responsible genotypes and to fuse the isolated protoplasts of Brassica carinata and Brassica napus breeding lines (BC DH Dodolla, BC DH -1, BC DH-6 and BN OP-1, BN-SL03/04), obtained from our previous experiments. Combination of three different PEG concentrations (20%, 25% or 30%) and two different treatment durations (15 and 20 min.) were tested. Our experiments identified several genotypes (Brassica carinata DH BC-6 and DH BC-1, Brassica napus DH OP-01) with satisfactory regeneration ability of calli from protoplast cultures. Proper combinations of concentration and treatment time of PEG determined protoplast fusion frequency between genotypes used. Although the 30% PEG solution was evaluated to be the best concentration, large amount of multifusants, unwanted in practical applications, was detected especially in Petri dishes with longer PEG treatment. In general, 25% PEG combined with 20 minutes treatment duration produced satisfactory fusion frequency and good rate of viability was obtained as well.