Induced mutation of curcuma hybrid cv. sweetmemory through tissue culture by ethyl methanesulphonate (EMS)

Abstract

The ethyl methanesulphonate (EMS) is induced mutation in Curcuma hybrid cv. sweetmemory. The shoot size 1 cm were cultured on Murashige and Skoog medium (MS) supplemented with 2 mg/l BA for plantlet induction. After 8 weeks of culture, it was observed that the shoots soaked in 0% EMS at 60 and 120 minutes survived 100%. In additions it was found the highest percentage of shoot emergence, number of shoots, and plant height. While plants treated with EMS solution had decreased survival and growth rates. Thereafter, existing plantlets were potted, and growth was observed. It turned out that the plants none receiving EMS solution (control) had the survival rate and growth rate were better than those of the plants receiving EMS solution. Results showed that control plants receiving 0% EMS at 60 minutes gave highest content of chlorophyll a as 4.06 µg/cm2, chlorophyll b of 2.08 µg/cm2 and carotenoid of 3.09 µg/cm2 but there was none statistical difference with the plants that received EMS solution. The control plants had larger stomata sizes than those that received the EMS solution. However, there was no difference in the number of chloroplasts among treatments. The living plantlets from all treatments were tested Random amplified polymorphic DNA (RAPD) marker and genomic DNA was extracted from fresh leaves. A dendrogram was constructed polymorphic bands using the NTSYSpc program (version 2.10p) showing 2 clusters which separated with similarity coefficients ranging from 0.74-1.00.