Defined Medium for Crown Gall Cells of Tobacco in Suspension Culture

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Abstract
The nutritional requirements of tobacco crown gall cells (auxin non-requiring cells) in suspension culture were investigated. In the basal medium (Murashige-Skoog inorganic medium), the crown gall cells did not disperse into suspension, growing in various sizes of firm cubic masses of cell aggregates. Individual components of yeast extract which is very effective for both cell growth and dispersion, were added to the basal medium and their effects on cell growth and dispersion were examined. Thiamine was effective for cell dispersion, and myo-inositol and nicotinic acid remarkably promoted cell growth. In the basal medium supplemented with these three components, the crown gall tissue grew rapidly being dispersed into a good suspension. An increase of inorganic phosphate in the medium was also very effective for cell growth.