In vitro mutagenesis of rose with ethylmethanesulphonate (EMS) and early selection using RAPD markers

摘要

In vitro-grown meristems of Rosa hybrida cv. First Red was used with various concentrations [0, 0.5, 1.0, 2.0 and 3.0% (v/v)] of ethylmethanesulfonate (EMS) to develop mutants. Depending on the concentration used, 50% survival was obtained in microshoots pretreated with 1.0% (v/v) EMS for 6 hours. The treated microshoots were used for further shoot multiplication. The maximum rate of shoot multiplication was obtained on MS medium supplemented with various concentrations and combinations of 2.0 mg/l BA (6-benzylaminopurine), 0.25 mg/l NAA (1-napthalene acetic acid), 100 mg/l Ads (adenine sulfate), 0.5 mg/l GA(3) (gibberellic acid) and 1.0% (v/v) EMS. The elongated shoots were rooted on half-strength MS medium supplemented with 0.25 mg/l IBA (indole-3-butyric acid) within two weeks of culture. About 70% of rooted plantlets survived in the green house. The genetic variation was detected among the in vitro raised plants by using RAPD markers. The result showed that 21.5% polymorphism was obtained among the in vitro raised plants. The in vitro raised mutants, as well as control plants, were grown in the pots. There was morphological variation between mutant and control plants, with regard to plants height, number of branches, number of flowers, stalk length of the flower, leaf size and number of petals/flower. This study will be of help for developing mutants for rose improvement programs.