Aseptic Conditions for Plant Tissue Culture

Aseptic Conditions for Plant Tissue Culture:

The sugar content of the nutrient media may support a luxuriant growth of many micro­organisms, like bacteria and fungi. It is, therefore, extremely important to maintain a completely aseptic environment inside the culture vials.

Micro-organisms can contaminate the medium in at least three ways:

(a) The micro-organisms present in the medium right from the beginning many be destroyed by sterilizing the properly plugged culture vials. It can be done by maintaining the temperature at 120°C for about 15 minutes.

(b) The micro-organisms may also be carried along with tissue that is being cultured. To prevent this, the plant material from which the tissue is to be excised is surface sterilized. The material may be surface sterilized with saturated chlorine water and then thoroughly washed with sterilized distilled water and to remove all traces of chlorine. If the material is fairly hard, as are some fruits and seeds, it may be surface sterilized by rinsing in alcohol.

(c) Finally, precautions must also be taken to prevent the entry of micro-organisms while the plug of a culture vial is removed to transfer the tissue to the nutrient medium (inoculation) For this, all operations from surface sterilization of the tissue up to inoculation are done in an aseptic environment.

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Research Paper # 4. Aeration Required for Plant Tissue Culture:

Proper aeration of the cultured tissue is also an important aspect of culture technique. If the tissue IS grown on the surface of a semi-solid medium it acquires enough aeration without special device for liquid medium, special device “filter paper bridge” is used. Here, two legs remain medium.