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egenerat~on from protoplasts and callus cultures of Hedysarirnr coro~~ariur~i were
optimized (.4rcioni et a., 1985). Protoplasts were ~soiated tkoni ~~icsopi~ylls of Mcdicago
sari\,a and various phytohormoncs tested for a better frequency of regeneratloll (KIIII and
Cho~, 1989). Frotoplasts of Lolus coniiculurus were ~solated us~t~g pre-plasn~olysaiio~i of
green cotyledo~i, in CI'W salts containing 13% iliann~tol. Plantleta were also regeiicrated
w~tll two lines being somaclo~ial varlanta (Vessabulr atld Grant, 1905). Callus and
protoplast CUI~LI~SS were used to Iregenerote ~)ln~lilcts ol"Wedinigu and leaves of hleilicagu
Iirroralb, an anlllial legunic reslstat~t to li~llgl~s l's~'i~i/op~'~i~n ~tii'di~~giiris. I'lalltlets wcrc
regeiicrated 011 niedium co~ltai~ii~ig 2-IP cornbi~icd wit11 IAA, andlor DAP w~th NAA (ZaSar
et al., 1995). Pla~lt regoleration fium cotyledo~l p~oloplasls was achlcvcd in Meiiicagu
saiiva cv. Krnsnovodopadskaya 8 by culturing cell aggregates on agar tiiedium aftcr tlic
immobilizat~on of protoplasts in agarose. Regeneration was also achieved from tissue
cultures of tile wild species M. prosrrara, hi, urbiculuris, M. rruurverleri, M. borealis, M.
cueriilea, M. rigiduila and M fulcnrn (Svanbaev, 199 1) Cotyledon protoplasts served as
useful tools for regeneration of Sesbatiia bispi~losa. 111 a liquid-over-agar culture systetn
with MS ~nediutii supplemented with 2,4-D, BAI', glutanline and mannitol, 84% of these
protoplasts divided and formed callus. Callus fosn~ed from the protoplasts differet~tiated
shoots on MS medium supplemented wit11 IBA arid BAP. These slioots developed into
complete platitlets when excised and cultured on MS containing IBA (Zhao et al., 1995).
Protoplasts der~vcd froin 3 species of Siylosarrrl~es were cultured in K81' medium at
densities of 5 X 1041t1iI and I X 1051tiil I'rotoplast-derived colonies tratisferred to MS
tiiediu~n suppleme~ited wit11 cot~ibitiatiot~s of NAA and BAP formed compact, green
microcalluses. Shoot regeneration, which occurred after 28-56 days of culture, was by