KLÍMA M. , ABRAHA E., VYVADILOVÁ M., BECHYNĚ M.
Abstract
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-SL-03/04),
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.
Key words:
Brassica carinata, Brassica napus, protoplast culture, protoplast fusion