Establishment Of In Vitro Regeneration System Of Fudingdabaicha

This experiment takes stems and leafs of Fudingdabaicha as explants to establish a set of vitro regeneration system.The results go as follows:(1)In order to reduce the frequency of contamination and browning of explants,the best time to drawing materials is from April to May.(2)Taking tender stems of Fudingdabaicha as explants,the optimum sterilization way is following:tender stems without leaves;dipped in the detergent water about two to three minutes and used a soft brush to clean the surfAce dirt;washed one point five or two hours by tap water;putted on the extra-clean desk;dipped in the 75%alcohol about 25-30s;washed 2 times by aseptic water;dipped in the 0.1%HgCl2 4min;washed two times by aseptic water;dipped in the 0.1%HgCl25min;washed four to six times by aseptic water;water dried and inoculated.The contamination rate of explants was 53.33%;the browning was 10%.(3)Drawing materials in April or May,the explants should be putted in lower temperature(15?)and dark condition for 3 days after inoculation,then put them in normal light and temperature to culture,doing like this can reduce the rate of browning of explants.(4)In primary culture process of Fudingdabaicha,phenomenon of explants contamination is serious.Several antibiotics or antiseptics such as ampicillin,streptomycin sulphate and Yipeiling were used in order to control the explants contamination of Camellia sinensis.The results showed that the effect of using single antibiotic on was worse than the combination.When using a single antibiotic,the effect of using streptomycin sulphate was better than the ampicillin.The effect of using Yipeiling is the worst.The culture medium adding 200mg·L-1 ampicillin and 150mg·L-1 streptomycin sulphate had contamination rate of 36.67%.(5)The tender stems and leaves were used as explants to induce callus,the result showed that induction effect of tender stems was better than leaves.(6)The best medium for inducing buds was MS+1.5mg·L-1 6-BA + 0.3 mg·L-1 NAA;The best medium for inducing callus was MS+2.0mg·L-16-BA+1.0 mg·L-1NAA,induction rate is 100%.Induction effect of NAA to callus was better than 2,4-D.MS was the best medium for callus growth and the optimum hormone combination was 6-BA1.0 mg·L-1 +NAA0.3 mg·L-1 +KT0.3 mg·L-1.The biggest proliferation rate was 313.33%.The optimum medium which induced adventitious bud proliferation was MS+1.0 mg·L-1 6-BA+0.1 mg·L-1 NAA.(7)The optimum medium which induced root was 1/2MS+NAA0.2 mg·L-1,the rooting rate was70%.The average root number was 4.0 and the average length of root was 5.4cm and the transplanting survival rate of plantets was 71.4%.