PLANT REGENERATED FROM MESOPHYLL PROTOPLASTS OF PINELLIA-TERNATA BREIT

Large numbers of viable mesophyll protoplasts are isolated from soil-grown plants of Pinellia ternata Breit of Araceae by liquid-solid double layer media with different concentrations of mineral salts, hormones and sugar. The first division of protoplasts takes place in 4-7 days of culture, with a division frequency of 3-8%. Cell clusters of 80-100 cells form in 3 weeks. They are transferred into a liquid medium to culture for a month on a shaker to form calli of 1-2 mm diameter which are transferred onto a solid differentiation agar medium. The calli first proliferate and grow, and differentiate into green buds and small seedlings in 3-4 weeks. The plantlets of Pinellia ternata regenerated from protoplasts grow to a length of 6-10 cm in another month. It was found that the shoot-forming process in the calli was to proceed in two ways: one was the organogenesis way, and the other was the embryoid way. Comparisons were made between the effects obtained from the 3 different methods, i.e. component concentration difference liquid-solid double layer culture, Iiquid-solid double layer culture, and liquid-suspension culture, for culturing protoplasts; and discussions were held.