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Tissue Culture of Annual Ryegrass x Tall Fescue F₁ Hybrids: Callus Establishment and Plant Regeneration¹

Calluses were established from internode and peduncle explants of field-grown annual ryegrass (Lolium multiflorum Lam.) x tall fescue (Festuca arundinacea Schreb.) F₁ hybrids, and plants were regenerated from the calluses. Modified MS medium with different levels of 2,4-dichlorophenoxyacetic acid (2,4-D) was used to initiate (2 to 4 mg of 2,4-D/liter) and maintain (2 mg/liter) callus and to regenerate (0.25 mg/liter) plants. Plants regenerated after the first two subcultures at 3 to 4-week intervals appeared to be identical in appearance and chromosome number to the original source plants. The doubled chromosome number was obtained in some plants regenerated after an 8-week subculture period in which the callus approached senescence. Increasing the number of subcultures before regeneration increased the frequency of phenotypic variants among the regenerants. The techniques described offer an opportunity for forage grass modification via cell and tissue culture.

Key Words: Forage grasses • Lolium multiflorum • Festuca arundinacea • Plant cell and tissue culture

² Plant physiologist and research agronomist, SEA-AR-USDA and Dep. of Agronomy, Univ. of Kentucky, Lexington, KY 40546; and professor, Dep. of Agronomy, Univ. of Kentucky, Lexington, KY 40546.

Received for publication January 29, 1979.