The plants and tubers of E. nuda were collected in rainy season
(July–August) from Mount Abu’s ranges in Sirohi district of
Rajasthan (India). These materials were brought to laboratory,
grown and maintained in the greenhouse conditions. Tubers were
harvested and cleansed to remove adherent soil. Cut-tubers (about
1 cm in size) were used as explants. Explants were pre-treated with
0.1% (w/v) each of streptomycin (antibiotic) and bavistin (a systemic
fungicide; BASF Ltd., India) for 15–17 min followed by 6–8
times washing with sterile water. Explants were surface sterilized
with 0.1% (w/v) HgCl2 (disinfectant, Hi-Media, India) for 5–7 min
under the aseptic condition in a laminar air hood followed by thorough
washing with sterile water 6–8 times. The surface sterilized
tubers explants were finally dipped in 70% ethanol for 60 s and
allowed to air dry.