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Embryogenic Suspension Culture and Protoplast Islolation in Litchee

Yu Changhe;Chen Zhenguang (Department of Horticulture, Fujian Agricultural University, Fuzhou 350002)  
Friable,granular embryogenic calli of litchee (Litchi chinensis Sonn ) were transferred into liquid medium for culture,and suspension cultures were established after two to three weeks of culture. Long -term cultures (more than ten subcultures) resulted in reduced growth and embryogenic capacity, whereas alternative culture with liquid medium (six subcultures )and solid medium (one subculture )kept the suspension cultures in a good condition. Sources of embryogenic suspension cultures,subculture duration,concentrations of both enzyme and mannitol in isolation solution,and proplasmolyzing all had significant effects on protoplast isolation from embryogenic suspension cultures. The optimum process for isolating of protoplasts was developed. The 4th -day cells used were proplasmolyzed in 13 % mannitol for one hour,then macerated in enzyme solution containing 0. 8 % calllase R - 10, 0. 4 % Macerease R - 10and 10 % or 11 % mannitol. Protoplasts were repetitively isolated at a yield of over 107/gfw and viability of 92 % or more under the optimum conditions.
【CateGory Index】: S667
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