Preliminary Study On Tissue Culture Of Anneslea Fragrans Wall.

Anneslea tree (Anneslea fragrans Wall.) is a plant species belonging to Camellia family, which is one of local specialty plant family in China. Anneslea tree has become famous as an ornamental tree for gardens and parks in many countries. In recent years, the increase of regional developing projects has led to sharp decrease of this ancient and rare plant in naturally distributed habitats and natural population. And it is rated as the endanger plant in China.The research on tissue culture of Anneslea tree will provide a clew and advanced technology for protecting the Anneslea tree resources and its production. So it is very important not only for theoretical research, but also for application.We obtained the regenerative saplings from dormant buds, stems and leaves through the direct as well as indirect way. The factors that affect every step including basic medium, hormone and active carbon are detailedly investigated. As a result, the optimum mediums of every step are screened. A plant regeneration system of Anneslea fragrans Wall. was established in this research.Orthogonal design was adopted on initial culture, and the buds were used for inducing sprout of top buds, the leaves and stems were used for inducing callus. The results show that the medium MS+1.0mg·L-1BA+0.1mg·L-1NAA is the optimum medium for sprouting of top buds. Both explants from open air and sterile laminas and leafstalks of regenerative plantlets can be used for inducing callus, and optimum medium are MS+2.0mg·L-1BA+0.5mg·L-12,4-D+0.1mg·L-1NAAandMS+ 2.0mg·L-1BA+2.0mg·L-1NAA.The optimum medium for proliferation are concluded by L9(33) orthogonal experiment. The medium MS+BA2.0mg·L-1+TDZ0.1mg·L-1 is propitious to proliferation of buds, and the medium MS+1.0mg·L-1BA+0.5mg·L-1NAA+ 0.01mg·L-1TDZ is propitious to proliferation of callus, the callus could proliferate fast on this medium.The optimum medium for inducing sprout of axillary buds is MS+2.0mg·L-1BA, which is concluded by L16(44) orthogonal experiment. This medium can be used for the explants from open air also, the ratio of inducement could be up to 63.4%. The optimum medium for inducing differentiation of callus is MS+2.0mg·L-1BA+ 0.05mg·L-1TDZ+0.1mg·L-1IBA, which is concluded by L9(33) orthogonal experiment.WPM+1.0mg·L-1BA+1.0mg·L-1NAA is the optimum medium for stronger nursling culture. On rooting culture, we concluded that the WPM+1.0mg·L-1NAA+0.5g·L-1AC is the optimum medium for rooting culture for the preliminary study.