| On the basis of provenances investigation of Phellodendron amurense Rupr., the seeds of Phellodendron amurense Rupr. from Dailing provenances because.of the highest content of the berberine among the trees investigated were chosen asthe origanl material of tissue culture, and we analysed the factors of micropropagation technique and callus culture of Phellodendron amurense Rupr.. And the content of berberine in callus cultured in different conditions were compared. The results were as follows:1. Through the comparison of 5 kinds of culture medium, we found that MS was compatibly for callus induction of Phellodendron amurense Rupr., phytohormones 2,4-D, NAA were all advantageous to callus induction, but higher concentration of phytohormones was inhibiting for callus formation. And the induction effect of NAA cooperated with 6-BA was better. Stem explants inducted by MS added 0.3 mg-L-16-BA+2.0 mg.L-1NAA growing the best with 90% induction rate and the combination were suitable for subculture. The most suitable pH for callus induction and growth was around 6.3, and the carbon sources concentration was 20 g.L-1; The optimal length of stem explants was 0.7 cm's among 0.3-1.0cm of them, and the best inoculation way of explants among the three ways(level-placing, straightcut,inserting)was level-placing on the media surface.2. The callus culture technique of Phellodendron amurense Rupr.: When the callus was subculture on the MS medium, it browning badly, while NT medium was suitable for its growth. When NAA was used only, the best combination was NT added 1.0mg·L-1 NAA; when worked with 6-BA, the combination of NT +0.3 mg·L-16-BA+1.0mg·L-1NAA was the most conduced to callus growth with the increase rate of 392%, and under that condition its growth was obviously bigger than that using NAA only.3. The system for berberine accumulation in callus was established: The highest content of berberine was obtained when cultured for 3 weeks and subeultured for 4 times in the NT medium add 0.3mg.L-1 6-BA+1.0mg·L-1 NAA with pH 6.3 and carbon sources concentration 20g·L-1, and the highest content was 2.14%.4. The-micropropagation technique of Phellodendron amurense Rupr. was also established: Adventitious buds were inducted for the stem with axillary buds cultured in MS+0.5mg.L-16- BA+0.1-0.2mg.L-1 NAA; and the differentiation frequency of adventitious buds was as high as 92.6%; the average number of them was 6.7. the shoots rooting in the WPM medium added 1.0 mg·L-1IBA and the average rooting rate was 60%. The suvive rate of transplant was up to 85%.
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