Plant Tissue Culture

Plant tissue culture is a collection of techniques used to maintain or grow plant cells, tissues or organs under sterile conditions on a nutrient culture medium of known composition. It is widely used to produce clones of a plant in a method known as micropropagation.

A whole plant can be regenerated from a small tissue or plant cells in a suitable culture medium under controlled environment. The plantlets so produced are called tissue-culture raised plants. These plantlets are a true copy of the mother plant and show characteristics identical to the mother plant.

Plant tissue culture can be initiated from almost any part of a plant however, for micro propagation or direct shoot regeneration, meristematic tissue such as shoot tip is ideal

Establishment of Commercial Plant Tissue Culture Unit

Storage room for chemicals:

It is advisable to have a separate area for storage of chemicals, apparatus and equipment. Chemicals required in small amounts should not be purchased in large quantities as they may lose their activity, pick up moisture or get contaminated. Such problems can be overcome by purchasing small lots on a regular basis

Washing and Media Preparation Room:

The glassware washing area should be located near the sterilization room. This area should have at least one large sink but two sinks are preferable with running tap water. Adequate workspace is required on each sides of the sink; this space is used for glassware soaking and drainage. The media preparation room should have smooth walls and floors, which enable easy cleaning to maintain a high degree of cleanliness. Minimum number of doors and windows should be provided in this room but within the local fire safety regulations. Media preparation area should be equipped with both tap and purified water.

Inoculation Room:

The most important work area is the Inoculation room where the core activity takes place. The transfer area needs to be as clean as possible with minimal air disturbance. The walls and floors of the Inoculation room must be smooth to ensure frequent cleaning. The doors and windows should be minimal to prevent contamination but within the local safety code. Sterilization of the instruments can be done with glass-bead sterilizers or flaming after dipping in alcohol, usually ethanol. Special laboratory shoes and coats should be worn in this area. Ultraviolet (UV) lights are sometimes installed in transfer areas to disinfect the room; these lights should be used only when people and plant material are not in the room.

Growth Room:

The culture room is an equally important area where plant cultures are maintained under controlled environmental conditions to achieve optimal growth. It is advisable to have more than one growth room to provide varied culture conditions since different plant species may have different requirements of light and temperature during in vitro culture. Also, in the event of the failure of cooling or lighting in one room, the plant cultures can be moved to another room to prevent the loss of cultures. In the growth room, the number of doors should be minimal to prevent contamination. The primary source of illumination in the growth room is normally from the lights mounted on the shelves.

Plant Tissue Culture