SPONTANEOUS SHOOT ORGANOGENESIS AND PLANT-REGENERATION FROM HAIRY ROOT CULTURES OF HYOSCYAMUS-MUTICUS

A rapid procedure for efficient plant regeneration from hairy root cultures of Hyoscyamus muticus L., transformed with Agrobacterium rhizogenes strain A4, is described. Spontaneous shoot organogenesis was observed in hairy root cultures after 3 subculturing passages in liquid LS0 medium. Shoot multiplication could be enhanced by transferring the shoot-producing roots to solid LS0 or B50 medium. Shoots formed roots on B5 medium supplemented with different combinations of auxins as well as on B50 medium. Hairy root tissue and regenerated plant material contained Ri-T-DNA. A large number of plants could be regenerated within 6-8 weeks. The plants differed phenotypically from normal plants in having wrinkled, narrower leaves, reduced apical dominance and abnormal flowering. The hyoscyamine content of the regenerated plants reflected the same pattern as in the control plants. The foliar hyoscyamine content increased during the growth period, reaching its maximum (1.15% of dry wt.) at the full flowering stage.