| Eucommia ulmoides’Hongye’is a varieties of Eucommia ulmoides Oliver in Eucommia Oliver of Eucommiaceae which has the important ornamental value and economic value, and has great prospects for development. Grafting reproduction is the main breeding way of Eucommia ulmoides’Hongye’, which required a lot of materials and has a long breeding cycle. That restricted the Eucommia ulmoides’Hongye’factory production and application. This paper try to use tissue culture method to reproduce Eucommia ulmoides’Hongye’The experiment take two years old Eucommia ulmoides’Hongye’as materials. Have research on effects of the variety of disinfectant and sterilization time to the proliferation rate of exophyte. Have research on influence of exophyte type and sampling time, the basic culture medium, exogenous additiveskind, the concentration and variety of hormone on the induction rate of shoot clumps and callus. Carried out shoot clumps induction test, induction and shoot clumps proliferation test of callus. Had the following main results:1. The types of disinfectant and sterilization time have a significant impact on survival rate of exophyte. Use 75% C2H6O and 0.1% Hgcl2 is the suitable sterilization method of Eucommia ulmoides’Hongye’. Suitable sterilization time and survival rate are different depending on the type of exophyte. Suitable sterilization method for terminal bud and leaf is 75% C2H6O soak 45 s and 0.1% HgCl2 soak 3 min, the survival rate of leaf is 88.89%, the survival rate of terminal bud is 22.23%. Suitable sterilization method for stem and stem with a bud is 75% C2H6O soak 45 s and 0.1% HgCl2 soak 5 min,the survival rate of stem with a bud and stem is 66.67%.2. The types of Eucommia ulmoides’Hongye’exophyte have a significant impact on the shoot clumps induction rate and callus induction rate. Sampling parts of exophyte has no significantly impact on shoot clumps induction rate and callus induction rate. Stem with a bud is the suitable exophyte for shoot clumps induction, shoot clumps induce from terminal bud are easily browning,stem and leaf cannot induce shoot clumps. Suitable exophyte for callus induction is leaf.3. The type of basic medium have a significant effect on the induction of callus and buds. Suitable basic medium for Eucommia ulmoides ’Hongye’buds and callus induction is MS medium. Shoot induction rate of stems with a bud on MS medium is 56.67%, callus induction rate of leaf is 90.0%.4. Sucrose concentration of 30 g/L, the highest shoot induction rate is 56.67 percent, the highest callus induction rate is 90%. Activated carbon can significantly reduce the browning rate of stems with a bud, concentration of activated carbon of 1.0g/L, the lowest buds browning rate is 10.0%, the highest shoot induction rate is 63.33%.5. Hormone types and concentrations have a significant effect on shoot and callus induction:6-BA can significantly promoting induction of stems with a bud, NAA and GA3 have significant inhibition on stems with a bud inducing,6-BA concentration of 1.0 mg/L, the highest shoot induction rate was 86.67%. NAA/6-BA is 2, induction of the stems with a bud and callus is 100%.6. Exophyte sampling time has a significant effect on the induction rate of buds and callus of Eucommia ulmoides’Hongye’. The best sampling time is March which has highest induction rate of buds and callus. Shoot induction rate in order are:March> April> May> September> July = November; callus induction rate in order were:March> April> May> September> July> November.7. Suitable buds proliferation medium is:MS+6-BA 1.0 mg/L, the multiplication factor of buds is 189%. Suitable callus subculture medium is:MS+6-BA 0.5 mg/L+ NAA 2.0 mg/L, the multiplication factor of callus is 243%. Suitable medium of buds induction from callus is:MS+6-BA1.0 mg/L, inducing rate is 33.33%. |
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