EMS induced point mutations in 18s rrna gene of hyoscyamus niger l. an important medicinal plant of kashmir himalaya

Hyoscyamus niger contains medicinally important tropane alkaloids such as scopolamine and hyoscyamine. In vitro mutagenesis, in combination with molecular marker methods, is a potent technique to produce genetic variability in medicinal plants. The objective of the study was to induce variation in In vitro raised H.niger using Ethyl Methane Sulphonate (EMS) as a mutagen. For this purpose the callii were treated with 5 different doses of EMS (0.01%, 0.02%, 0.03%, 0.04% and 0.05%) at different time intervals (1/2 hour and 1 hour), and the effect of EMS was observed during morphogenesis. The analysis rvealed that EMS had a profound effect on shoot number and length as well as percentage of shoot formation. 0.03% EMS was seen to be effectively enhancing the average shoot number from 13.5 to 22.0, with 100% callusing response under normal growth regulators. The 18S rRNA gene of the affected plantlets was then amplified and sequenced to observe nucleotide variations, including transitions and transversions, at different concentrations of EMS. (author)