Plant Tissue Culture/Micropropagation

Plant cells can be grown in isolation from intact plants in tissue culture systems. The cells have the characteristics of callus cells, rather than other plant cell types. These are the cells that appear on cut surfaces when a plant is wounded and which gradually cover and seal the damaged area.

Pieces of plant tissue will slowly divide and grow into a colourless mass of cells if they are kept in special conditions. These are:

• initiated from the most appropriate plant tissue for the particular plant variety
• presence of a high concentration of auxin and cytokinin growth regulators in the growth media
• a growth medium containing organic and inorganic compounds to sustain the cells
• aseptic conditions during culture to exclude competition from microorganisms

The plant cells can grow on a solid surface as friable, pale-brown lumps (called callus), or as individual or small clusters of cells in a liquid medium called a suspension culture. These cells can be maintained indefinitely provided they are sub-cultured regularly into fresh growth medium.

Tissue culture cells generally lack the distinctive features of most plant cells. They have a small vacuole, lack chloroplasts and photosynthetic pathways and the structural or chemical features that distinguish so many cell types within the intact plant are absent. They are most similar to the undifferentiated cells found in meristematic regions which become fated to develop into each cell type as the plant grows. Tissue cultured cells can also be induced to re-differentiate into whole plants by alterations to the growth media.

Plant tissue cultures can be initiated from almost any part of a plant. The physiological state of the plant does have an influence on its response to attempts to initiate tissue culture. The parent plant must be healthy and free from obvious signs of disease or decay. The source, termed explant, may be dictated by the reason for carrying out the tissue culture. Younger tissue contains a higher proportion of actively dividing cells and is more responsive to a callus initiation programme. The plants themselves must be actively growing, and not about to enter a period of dormancy.

The exact conditions required to initiate and sustain plant cells in culture, or to regenerate intact plants from cultured cells, are different for each plant species. Each variety of a species will often have a particular set of cultural requirements. Despite all the knowledge that has been obtained about plant tissue culture during the twentieth century, these conditions have to be identified for each variety through experimentation.