| Litchi (Litchi chinesis Sonn.), a member of the Sapindaceae family, originated in china and it is a famous subtropical evergreen fruit trees in south of china. In vitro culture of litchi are very difficult, therefore, development of biotechnology in litchi is limited. In this research, anther of different species of litchi was inducted in an effort to obtain embryogenic callus. The system of high frequency somatic embryogenesis of litchi was improved; the system of plant regeneration of litchi protoplast was established and improved. In order to provide theory foundation and technology support for biotechnology breeding etc of litchi. The main results of this research were as follows.1. Sixteen species anther and different types of explants from similar species were inducted, obtained embryogenic callus from nine species of sanyuehong, feizixiao, shuidong, xiangshanjizuili, sanshanjianyeli, huaizhi, gualv, hongli, chengtuo etc, and obtained non-embryogenic callus from eight species, obtained embryogenic callus from young embryo of feizixiao, obtained non-embryogenic callus from stem and young leaves of sanyuehong. Found that the medium of MS+2mg/L 2, 4-D +0.5mg/L NAA (2.0mg/LKT)+500mg/L PVP +50g/L sucrose adapted to embryogenic induction of anther and young embryo of litchi.2. The results of experiment indicted that embryogenic callus could multiplication very well in the medium of MS+2mg/l 2, 4-D +0.2mg/L NAA +30g/L sucrose. In order to maintain the embryogenic callus very well, a liquid-solid alternative culture method was followed: after five to six weekly subcultures in liquid medium, suspensions were transferred to solid medium for two weeks. Use this method, callus that subculture ten weeks transferred to the medium of non-plant growth regulator could produce a lots of somatic embryos. Culture in darkness could avoid callus brown in a certain extent. So far, embryogenic callus of shuidong, feizixiao, gualv, huaizhi, sanshanjianyeli, xiangshanjizuili had been preserved.3. The mediums that adapted to somatic induction of litchi embryogenic callus were selected by using the orthogonal experiment design. The medium is MS+0.5mg/L KT +0.1mg/L NAA +3g/L AC +30g/Lsucrose. In this medium, somatic embryos of feizixiao and shuidong had been obtained successfully. In order to obtain identical growth and little abnormality somatic embryos, a liquid-solid alternative culture method were performed. The method is cultured in liquid medium two weeks then transferred to solid medium to induce. At last, a lot of normal somatic embryos were obtained. At the same time, the results of experiments suggested that somatic embryo induction of litchi has relation togenotype.4. Regeneration plantlet could obtain from maturation somatic embryo in the medium of non-plant growth regulator. At present, regeneration plantlets of shuidong and feizixiao had been obtained. Maturation culture could accelerate somatic embryo developing and enhance the amount of regeneration plant. In the maturation medium, add to 500mg/Lglutamine, 400mg/Ljelly, lOOml/L coconut water, SOOmg/Lcasein hydrolyzate, the rate of plant regeneration enhanced relativity. Combination of different concentration of BA, GA and NAA and their effect on plant regeneration were probed into by using the orthogonal experiment design, but the result was not satisfied.5. Embryogenic suspensions that subculture four to five times and suspension culture four to six days was the best material from that protoplasts were isolated. 1.0% Cellulase Onzuka R-10 and 0.5% Pectolysae Y-23 were the best combination in protoplast isolation. Protoplasts of litchi in the medium of MB+l.Omg/LZT +1.0mg/L2 , 4-D+0.2mg/LNAA+500mg/Lglutamine+0.45mol/Lglucose+0. Imol/L sucrose could divide constantly and came into being microcolonies. Callus originated protoplast culture had been obtained in sanshanjianyeli and xiangshanjizuili.6. In this experiment, the multiple of embryogenic callus and regeneration plant of di...
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