The Technology Research For Tissue Culture Of Cyclobalanopsis Gilva (Blume) Oerst

Cyclobalanopsis gilva (Blume) Oerst, species of fagaceae is barren resistant and widely distributed. Wood of Cyclobalanopsis gilva (Blume) is of high quality and wide range of uses. It is one kind of valuable hardwood tree species in our country and has high economic value. Due to massive deforestation, it had become endangered species. On the one hand, the seeds are easy to destroyed by pests and hard to survive. At the same time, the sexual reproduction is slow. On the other hand, sexual reproduction can not guarantee excellent characteristics of forest trees. However, tissue culture as asexual method, can produce excellent characteristics of genetic female. In a short period, we can get a large number of quality tissue culture seedlings and provide a provenance promotion. In this study, seeds are come from Huaihua. The research content included the choice of explants and disinfection, basic culture medium choice, selection and concentration of hormone, callus induction, callus proliferation, callus differentiation, and induction of rooting and so on. The specific research contents and results are as follows:1. Sterilization of explants. For 1～2 years seedlings tender stem, the best disinfection treatment was 70% ethanol 15 s first, then 0.1% HgCl2 solution for 15 minutes. For the fresh seeds picked in current year, the best disinfection process was 70% ethanol treatment for 30s, then 0.1% HgCl2 for 10 min before peeling; 70% ethanol for 30 s and then 0.1% HgCl2 for 3 min after peeling.In those method, the pollution rate of seed was very low, and the germination rate was very high.2. Callus induction of explants and medium choice. Through experiment contrast analysis, bud was a better materials on callus induction; through a variety of basic culture medium contrast experiment, MS culture medium was selected as the best basic culture medium.3. Callus induction,proliferation and differentiation.6-BA and IB A asexperiment factors was studied. According to the results, the optimum medium was MS+1.0 mg/L6-BA+0.2mg/L IBA; Using different culture medium for callus proliferation and differentiation of the experiment, the best proliferation medium was MS+1.0mg/L6-BA+0.3 mg/L IBA; best differentiation medium was MS basic culture medium+1.5 mg/L6-BA and 0.1mg/LIBA.4. Bud induction and rooting. Bud induction included crown induction and cotyledonary induction for shoot clumps. Through the four factors three levels orthogonal experiment design and variance analysis, the result showed that the best medium for crown induction was improved 1/2MS+1.5 mg/L6-BA+0.01 mg/LIBA+ O.lmg/LTDZ; for cotyledonary node induced multiple shoot clumps was improved 1/2 MS as the basic culture medium by completely block experiment. The impact factor analysis of variance and Duncan multiple comparison the results finally indicated that the basic culture medium for multiple shoot clumps induction were improved 1/2MS+1.5mg/L6-BA+0.1mg/LTDZ. In terms of rooting induction,1/4 MS+1 mg/L6-BA+1 mg/LIBA worked best.