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Virus Symptom-Free Plants of Red Clover Using Meristem Culture¹

Recently developed techniques for in vitro red clover (Trifolium pratense L.) culture were adapted for the culture of shoot meristem tips. Symptomless plants were obtained from meristem cultures of the virus-infected clones of ‘Kenstar’ red clover. A basal culture medium designated ‘ML2’ which included 4.0 /µg/liter picloram (4-amino-3,5,6-trichloropicolinic acid) as the auxin and 1.0 mg/liter 6-benzylaminopurine as the cytokinin yielded single plants from excised shoot meristem tips. Higher auxin levels in the medium resulted in callus formation and multiple shoot initiation. Crown-derived meristems yielded normal appearing plants from culture while axilderived meristems produced abnormal plants. About 65% of the shoot meristem explants including the meristematic dome and first foliar primordium survived culture, and 80% of the larger meristem explants, including the second foliar primordium, survived culture. The smaller explants yielded a greater proportion of symptomless plants compared with the larger explants. About 90% of the meristem-derived shoots rooted on a medium designated R(PC). Tests using Chenopodium amaranticolor Coste & Reyn. as a virus bioassay plant suggested that twothirds of the meristem-derived plants were virus-free. Plants obtained in this manner will enable red clover breeders to maintain synthetic cultivars more efficiently, since seed set is greatly increased in virus-free plants.

Key Words: In vitro • Trifolium pratense L. • Virus-free plants • Asexual propagation

² Research assistant and professor, respectively, of the Dep. of Agronomy, Univ. of Kentucky, Lexington, KV 40546.

Received for publication September 7, 1978.