Study On The Tissue Culture Technique Of Buddleja Alternifolia

Buddleja alternifolia is a perennial deciduous shrub in the genus(Buddleja·L)of the family Marmonaceae(Loganiaceae).Its adaptability,extremely cold tolerant,drought tolerant and tiller,highly ornamental,good ecological habits and ornamental value.The growth conditions of wild wild fish grass are harsh,the natural emergence rate is low,the resources are decreasing,and it is urgent to establish a simple and efficient reproductive system.Tissue culture technology reproduces the seedlings quickly and is not affected by the external environment,so the seedlings can maintain the excellent characteristics of the mother strain.Tissue culture rapid propagation technology is an important way to meet the market demand for grass.Orthogonal test design was used to study the process of aseptic germination culture,stem segment induced axillary bud culture,different explants induced callus culture,leaf induced adventitious bud culture,rooting culture and seedling cultivation.The study results are as follows:(1)The optimal soaking temperature of seeds is 55℃;The optimal soaking time is16h;The optimal alkali erosion time of 40%Na OH solution is 15min;The best way to place it is it horizontally;The best disinfection method is to disinfect with 75% alcohol for 30 s,With 10%Na Cl O solution for another 22 min,At this time,the germination rate was 87.3%,The infection rate was 1.4%;The best formulation of the induction medium was MS+6-BA 0.6 mg/L + NAA 0.5 mg/L,Seed germination rate was 94.7%;The optimal proliferation medium culture formulation was MS+6-BA 0.8 mg/L+NAA 0.7mg/L+IBA 0.2 mg/L,The value-added coefficient was 6.73.(2)Stem-segment-induced axillary bud culture the best disinfection method was treated with 75%alcohol for 30 s and then 1%Na Cl O for 12 min,the survival rate was86.7%and infection rate was 3.3%;the optimal induction medium formula was MS+6-BA1.0 mg/L+NAA 0.5 mg/L,and the axillary bud induction rate was 91.7%;the optimal proliferation medium formulation was MS+6-BA 1.0 mg/L+NAA 0.2 mg/L+IBA 0.3mg/L and the proliferation coefficient was 6.47.(3)The best induc-tion medium formulation for both leaf and stem segments was MS+TDZ 1.0 mg/L+NAA 0.7 mg/L,Induction rates were 82.9%and 79.2%,respectively,Therefore,the best callus is leaf;The optimal proliferation medium formulation was MS+TDZ 1.0 mg/L+NAA 0.6 mg/L+6-BA 0.5 mg/L,The value-added coefficient is 4.39;The optimal differentiation medium was MS+6-BA 1.0 mg/L+IBA 0.3 mg/L,The differentiation rate was 84.4%,The average number of buds was 22.3.(4)The best induction medium for leaf-induced indefinite shoots was was MS+6-BA 1.0 mg/L+IAA 0.6 mg/L,with an induction rate of 87.2%;the best proliferation medium was MS+6-BA1.0 mg/L+IAA 0.7 mg/L+NAA 0.2 mg/L,with a proliferation coefficient of 4.27.(5)The optimal rooting medium for histone culture seedlings was 1/2MS+IBA 0.5mg/L+NAA 0.3 mg/L,and the rooting rate was 93.5%,and the average number was 7.7roots.(6)In the process of mutual leaves seedlings seedling management,the experimen-tal results show that refining seedlings to group seedlings open processing 1d,the seedlings gradually adapt to the external environment,and select the average root length between 3～5cm,then planted in the sterilized soil matrix for nutrient soil:vermiculite:perlite =4:2:1,seedling survival rate is 88.3%,in the process of management,select hole plastic film,keep the humidity between 50%-60%.The study found that the three culture methods have their own advantages,callus culture has good proliferation effect,but the overall culture time is longer,the blade is short,weak,and weak;the axillary bud culture has short time,robust growth and good growth.By comparing the three tissue culture methods,it was found that the best method of tissue culture.