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In Vitro Tissue Culture of Selected Legumes and Plant Regeneration from Callus Cultures of Red Clover¹

A synthetic, chemically defined medium was developed for the initiation and growth of cell and tissue cultures of red clover (Trifolium pratense L.). The medium is broadly supportive and suitable for the culture of other legume species, including alfalfa (Medicago sativa L.), soybeans (Glycine max L.) and jack bean (Canavalia ensiformis L). Cultural variables for red clover were defined, and it is now possible to routinely establish callus cultures of red clover from reproductive and vegetative explant tissues of both mature and immature plants. Cell suspension cultures with rapid cell division rates have been established from calli by adapting the new medium to liquid culture. Organogenesis was induced in callus cultures, and the recovery of whole plants regenerated from callus has been achieved with the cultivars ‘Altaswede’, ‘Arlington’, ‘Kenstar’, ‘Redman’ and ‘Tensas’, but they did not respond equally well. Regenerated plants have been grown to maturity and were fertile. 3-Aminopyridine was evaluated and found to possess growth-modifying properties. It is now possible to investigate the use of tissue and cell culture techniques in red clover breeding programs.

Key Words: Soybean • Alfalfa • Jackbean • Organogenesis • Cell suspension cultures • Tissue culture medium • growth regulators • Picloram • 3-Aminopyridine

² Research assistant and professor, respectively, of the Dep. of Agronomy, Univ. of Kentucky, Lexington, KY 40506.

Received for publication April 28, 1978.