Aim- Surface sterilization of plant material Requirements- Mold detergent Explant materials Sterile forceps Commercial bleach/ calcium hypochlorite Scalpels Ethanol(70%) Sterile distilled water Surfactant (Triton-X/Tween 20) Surface sterilization of plant material: Surface of the plant parts carry a wide range of microbial contaminants, fungi and bacteria being the common. To avoid the microbial growth which is, detrimental to culture growth therefore before inoculating the medium with the explants, it is necessary to surface sterilize it. There are many sterilizing agents such as calcium hypochorite, chlorine water, bleaching powder, mercuric chloride, hydrogen peroxide and ethylene oxide. The concentration of the sterilizing agent and the duration of treatment varies with the plant material. These should be no damage to the explants in the process of surface sterilization. The plant material should then be thoroughly washed with sterile distilled water before transferring it to the nutrient medium.

a) Sterilization of seeds: 1. About 15-20 healthy seeds were selected and wash them thoroughly with water. 2. Now added 2-3 drops of liquid detergent 100ml of water and shake well for 5 minutes. 3. Pour off the detergent solution and wash thoroughly to remove any traces of the detergent. 4. Rinse seeds in 50ml of 70% ethanol for 1 minutes. 5. Wash thoroughly 3-4 times with single distilled water. 6. Transfer seeds in a sterile 250ml flask and add mercuric chloride solution (0.1%). 7. Treat for some time, shake the flask occasionally. All the operations thereafter should be carried out under sterile conditions. 8. Transfer the flask to a sterile cabinet, decant the mercuric chloride solution and wash the seeds thoroughly with sterile water to remove any traces of mercuric chloride.