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Somatic Embryogenesis from Cell Suspension Cultures of Red Clover¹

In the present study, plants were regenerated from cell suspension cultures of ‘Arlington’ red clover (Trifolium pratense L.). Evidence was obtained which demonstrated that these plants regenerated via somatic embryogenesis. The source and concentration of auxin used in culture media had significant effects on the initiation of somatic embryogenesis. Somatic embryos were formed on up to 40% of the culture plates which contained 0.01 mg/liter 2,4-dichlorophenoxyacetic acid (2,4-D) (0.045 µM) and 2.0 rag/liter adenine (14.8 µM) in a basal dium. Shoots and plants were obtained on the same medium or after subculture onto medium containing 0.001 mg/llter 4-amino.3,5,6-trichloropicolinic acid (pi. cloram) (0.004 µM), and 0.2 mg/llter 6-benzyladenine (0.88 µM). Regenerated plants were characterized cyto. logically, morphologically, and reproductively. In vitro regeneration of plants is a prerequisite for the application of selection techniques at the cellular level. Genetic selection at the cellular level in red clover breeding should now be possible with the present identification of a reliable system for cell to plant totipotency

Key Words: Trifolium pratense L. • In vitro • Plant regeneration • Auxin effects

² Research assistant and professor, Dep. of Agronomy, Univ. of Kentucky, Lexington, KY 40546.

Accepted for publication August 15, 1979.