A Study On Callus Culture And Secondary Metabolites Of Eucommia Ulmoides Oliv.

The callus induction and subculture of Eucommia ulmoides Oliv. are studied from the aspects of basic media, installing method, type of explant, sampling time, culturing conditions, phytohormones etc. The influence of the condition of tissue culture on the contents of metabolites contained in the callus is also studied preliminarily. The results are as follows:1. Among the 8 media tested, MS is suitable for the callus induction of immature shoots of E. ulmoides Oliv. 65 medium however is suitable not only for callus induction of leaves, but also immature shoots.2. The callus are induced 100 per cent when explants are cultured on their medium supplemented with 0.5 mg/L 2,4-D, 0.5-1.0 mg/L NAA, 1.0 mg/L IBA is the best one for the induction and growth of callus. Combination of 2,4-D and BA or KT is worse than 2,4-D used singly. The combinations of low concentrations of BA and NAA or IBA have better effect, but high concentration of BA is not suitable. The best combination of the media for callus induction and growth is 1.0 mg/L NAA+0.3-0.5 mg/LBA or 1.0 mg/L IBA+0.5 mg/L BA. And calluses grow quickly with glossy appearance under this combination, and suitable for subculture. 0.5 mg/L NAA is the best one for callus subculture. The best combination is 0.5 mg/L NAA+0.5 mg/L BA. Their callus yield was higher obviously than using NAA singly. KT contains callus induction and growth.3. Middle of March is a favorable period for the collection of immature leaves as explant. Early April however is suitable for the collection of shoots. Other times are suitable for hypocotyl , cotyledon and leaf to culture sterile seedlings. The best pH value for the callus induction and subculture is 6.3. 12 hour light and 12 hour dark are benefit for the callus induction ,subculture and the production of secondary metabolites. 0.7 cm shoot explants implanted horizontally in the medium recommended.4. Sucrose as carbon source is better than glucose and refined sugar for callus induction. 40 mg/L sucrose is not only for callus subculture but also for the accumulation of flavonoids.5. Applications of KNO3, (NFL;) 2804, ZnSO4and FeSO4 can increase the growth of callus contains the of accumulation of chlorognic acid and flavonoids. MgSO4 is unprofitable for the callus growth or the accumulation of chlorognic acid and flavonoids. The applications of NaH2P04 and CaCl2 not only increase the callus growth, but also for the accumulation ofchlorognic acid and flavonoids. The applications of MnSO4 not only increase the callus growth and hlorognic acid accumlation, but not beneficial for the accumlation of flavonoids. The applications of sucrose can increase the growth of callus contains the of accumulation of chlorognic acid and flavonoids.