Plant Tissue Culture

What is Plant Tissue Culture?

Induction of onion tissue culture from onion seedlings (LHS), through callus tissue proliferating on seedling tissue to isolated callus in culture (RHS).

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.

PLANT TISSUE CULTURE Uses of plant tissue culture INTRODUCTION WHAT IS IT? USES CASE STUDY 1 Anther culture for cold hardiness CASE STUDY 2 Somaclones for disease resistance CASE STUDY 3 Demonstration of tissue culture for teaching PUBLICATIONS

Callus cultures derived from onion

Plant tissue culture now has direct commercial applications as well as value in basic research into cell biology, genetics and biochemistry. The techniques include culture of

cells, anthers, ovules and embryos on experimental to industrial scales, protoplast isolation and fusion, cell selection and meristem and bud culture. Applications include: • • • • • • •

micropropagation using meristem and shoot culture to produce large numbers of identical individuals screening programmes of cells, rather than plants for advantageous characters large-scale growth of plant cells in liquid culture as a source of secondary products crossing distantly related species by protoplast fusion and regeneration of the novel hybrid production of dihaploid plants from haploid cultures to achieve homozygous lines more rapidly in breeding programmes as a tissue for transformation, followed by either short-term testing of genetic constructs or regeneration of transgenic plants removal of viruses by propagation from meristematic tissues

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