Study On Technology About The Tissue Culture Of Camellia Oleifera

Camellia oleifera is an endemic woody edible oil plant insouthern China. In this study, We used improved variety of oleifera as material, with its immature embryos and cotyledon, the tender stems and leaves as the explants, selected the best medium for embryogenic callus, the best medium for various stages of formulation of induction, proliferation of cultured, rooting and transplanting for immature cotyledon, the tender stems and leaves, which were through tissue culture techniques and by the methods of orthogonal experiments, analysis of variance and multiple coparison. In addition, we explored the impact factor on the anther culture in Virto of Camellia oleifera, he main results are as follows:1. Study on tissue culture techniques of Camellia oleifera cotyledon and yong embryo. The best medium for aseptic Seedling was WPM+6-BA2.0mg/L +NAA1.5mg/L; the best medium for embryogenic callus was WPM+6-BA 1.0 mg/L+NAA0.5 mg/L; the best medium for embryoid inducton and bud differentiation was MS+6-BA 2.0 mg/L+NAA 0.01 mg/L2. Study on tissue culture techniques of Camellia oleifera tender stem segement. The best medium for bud induction was 1/2MS+6-BA1.0mg/L+NAA 0.5mg/L; The best medium for multiplication culture was 1/2MS+6-BA1.0mg/L +NAA0.05mg/L, and the proliferation times of multiple shoots could up to 5; The best medium for elongation was 1/2MS+sugar2g/L+NAA0.1mg/L. The rooting rate could up to 88.3% by the methods of shoots treatment in the solution of IBA1000ppm for 5s, and then turned into the 1/2MS blank medium for 30d. The shortest period which from the induction of buds to the obtaining of a plant only need 70d. For transplantion of plantlets, the best substrate was peat:vermiculite=1:1, with the highest survival rate.3. Study on tissue culture techniques of Camellia oleifera leaves. The best medium for callus inducing was WPM+TDZ1.5mg/L+NAA0.5mg/L+6-BA2.0mg/L; The best medium for redifferentiation was 1/2MS+TDZ1.0mg/L+NAA 0.1mg/L+6-BA2.0mg/L with the rate of 17.86%. The best medium for multiplication of buds was 1/2MS+6-BA1.0+ IBA0.05 mg/L; The rooting medium was 1/2MS+TDZ0.05mg/L+IBA1.0mg/L.4. Study of Impact Factor on The Anther Culture in Vitro of Camellia. The best method of low temperature pretreatment is keeping the branches of flower bud in the water; The time of low temperature pretreatment should be below 10 days; The best developing stage of pollen on inducing callus from oil-tea anther is tetrad stage; and the best medium additives is AgNO3, with the high induction rate (more than 80%) and a good morphology. In the culture of callus proliferation, the best recipe of callus proliferation culture medium is MS+2.0 mg/L 6-BA+0.05 mg/L NAA.