Study On The Technical System Of Tissue Culture And Rapid Propagation Of Clematis Lasiandra Maxim

The study was mainly to probe an efficient way through tissue and rapid propagation of Clematis lasiandra Maxim.The stems with buds,internode and the leaves of young branches were taken as explants. To find out the better medium and proceeding of culturing Clematis lasiandra Maxim., we used orthogonal experimental design and analysis of variance to establish the technique system of tissue culture. The affected factors of propagation, existing problems in this technique and the corresponding solutions were also described, which provide the theoretical base and technical measures for industrial producing of plantlet.The study of tissue culture technique and the better medium for various stages were as follows:①March were the best season to draw materials,in which the starting rate of explants was high while the pollution was low.The starting rate of the stems with buds,internode and the leaves were 81.67%,66.67% and 75.00% respectively;the pollutiong were 15.00%,22.03%,13.33%.②The best sterilization time for Clematis lasiandra Maxim by HgCl2(0.1%) was 5minutes to the stems with shoot,5minutes to the segment and 3minutes to leaves.③Initial culture:Initial medium of the stems with buds:MS+6-BA1.00 mg·L-1+ NAA0.10 mg·L-1 was suitable,in which the starting rate was 90.56%, and the shoot forming index was 1.75;The medium of callus induction for internode:MS+6-BA2.00 mg·L-1+NAA0.10 mg·L-1 was suitable,in which the callus induction rate was 83.30%. The medium of callus induction for leaves:MS+6-BA0.50 mg·L-1+NAA0.10 mg·L-1+ 2,4-D 0.10 mg·L-1 was suitable,in which the callus induction rate was 80.30%.④Multiplication culture:The optional culture medium combination for buds proliferation was:MS+6-BA 1.00 mg/L+NAA 0.10 mg·L-1, proliferation index was 3.2 after inoculating 30 day.The optional culture medium combination for calluses proliferation was:MS+6-BA 2.00 mg-L"'+NAA 0.10 mg·L-1+2,4-D 0.01 mg·L-1, the proliferation rate could be 24.45%; The optional culture medium combination for calluses diferentiation was:MS+6-BA 2.00 mg·L-1+NAA 0.05 mg·L-1, the differentiation rate could be 36.67%。⑤Anti-browning of calluses:MS+6-BA2.00 mg·L-1+NAA0.10mg·L-1+2,4-D 0.01mg·L-1+PVP 2 g·L-1 was better,which the browning rate was 30.43%.⑥Efficiency seeding culture:MS+suger 40g·L-1, in which the rate of efficiency seedings was 65.21%.⑦Rooting culture:1/4MS + NAA 0.50mg·L-1+AC1.00 g·L-1+suger15 g·L-1 in which the rate of rooting was 86.10%, the quantity of roots was 4.3.⑧Forging seeding transplanting:The transplant medium in perlite,vermiculite, and humus by 1:1:2 was the optimal for Clematis lasiandra Maxim., which the rate of survival was 75.00%.