Serial Number

Name of Media

Media composition and culture condition

Period of Culture

(weeks)

1

MS1 for semi-solid culture

MS basal medium ( Murashige & Skoog, 1962 ) supplemented with 2.5 mgl⁻¹ BA (6-benzyladenine), 0.5 mgl⁻¹ GA₃, 30 gl⁻¹ sucrose solidified with 7 gl⁻¹ agar and pH was adjusted to 5.7 - 5.8 and autoclaved it and dispensed 20 ml in sterilized condition in each sterilized 90 mm petri dish.

3

2

MS2 for semi-solid culture

MS salts, B5 vitamins, iron source, myo-inositol 0.1 gl⁻¹, sucrose 30 gl⁻¹, 2,4-D 2.5 mgl⁻¹, were mixed and made the volume 1 litre and adjust the pH 5.6 - 5.8 and add Agar 8 gl⁻¹ mixed and autoclaved it and dispensed 20 ml in each sterilized 90 mm petridish.

16

3

MS3 for suspension culture

MS salts, B5 vitamins, iron source, myo-inocitol 0.1 gl⁻¹, 2,4-D 2.5 mgl⁻¹, STS 29.4 μM and adjust the pH 5.6 - 5.8 and autoclaved it and dispensed 50 ml in each Erlenmeyer Conical flasks. In each flask friable callus was put, mixed and placed on 120 rpm agitated orbital shaker incubated at 25˚C ± 2˚C and sub-cultured in every 2 weeks.

8

4

MS4 for liquid culture

MS major and minor salts, vitamins, iron source, myo-inositol 0.1 gl⁻¹, BAP (6-benzylaminopurine) 20 mgl⁻¹, sucrose 30 gl⁻¹ were mixed and made the volume 1 liter and adjust the pH 5.6 - 5.8 and autoclaved it and dispensed 20 ml in each culture tube having filter bridge.

16