Development of in vitro procedures for regeneration of petiole and leaf expiants and production of protoplast-derived callus in Tanacetum vulgare L. (Tansy).

Tanacetum vulgare (Tansy) was established in vitro on Murashige and Skoog (MS) medium supplemented with αnaphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP) using shoot tips and embryos. From petiole expiants 93% formed callus, and 27% produced shoots on MS medium containing 4.5 mg l(-1) NAA and BAP. NAA alone induced root formation from leaf expiants. Up to 7 ×10(6) viable protoplasts were obtained by macerating 1 g of leaves in 0.5 % Macerozyme R-10, 1.0% Cellulase R10, and 1.0% Cellulysin. Cell division was observed 3-4 days after protoplast isolation at the optimum plating density of 0.2-0.4×10(6) cells ml(-1). A total of 350 protoplast-derived calluses were produced on which nodules with meristematic zones developed. Roots regenerated on MS medium supplemented with BAP 3.0 mg 1(-1), NAA 2.0 mg l(-1), and 250 mg l(-1) casein hydrolysate, however no shoots have been obtained yet.