DEVELOPMENT OF IN-VITRO PROCEDURES FOR REGENERATION OF PETIOLE AND LEAF EXPLANTS AND PRODUCTION OF PROTOPLAST-DERIVED CALLUS IN TANACETUM-VULGARE L (TANSY)

Tanacetum vulgare (Tansy) was established in vitro on Murashige and Sk oog (MS) medium supplemented with alpha-naphthaleneacetic acid (NAA) a nd 6-benzylaminopurine (BAP) using shoot tips and embryos. From petiol e explants 93% formed callus, and 27% produced shoots on MS medium con taining 4.5 mg l(-1) NAA and BAP. NAA alone induced root formation fro m leaf explants. Up to 7 x 10(6) viable protoplasts were obtained by m acerating 1 g of leaves in 0.5% Macerozyme R-10, 1.0% Cellulase R-10, and 1.0% Cellulysin. Cell division was observed 3 - 4 days after proto plast isolation at the optimum plating density of 0.2 - 0.4 x 10(6) ce lls ml(-1). A total of 350 protoplast-derived calluses were produced o n which nodules with meristematic zones developed. Roots regenerated o n MS medium supplemented with BAP 3.0 mg l(-1), NAA 2.0 mg l(-1), and 250 mg l(-1) casein hydrolysate, however no shoots have been obtained yet.