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为提高用植物细胞培养生产次生代谢产物的单位产量，作者对黄连细胞采用二步法悬浮培养生产黄连生物碱进行了探索。结果显示：黄连细胞采用一步法悬浮培养６周，细胞干、鲜重产率分别为２０．９６ｇ／Ｌ和１７４．９２ｇ／Ｌ，增重约７００％；细胞中总生物碱含量为１４．７９ｍｇ／ｇ．ｃｅｌｄｗ，培养基中生物碱量为１２．９６ｍｇ／Ｌ，每升培养液共可收获生物碱３２３ｍｇ。采用先在生长培养基中培养３周，然后在合成培养基中培养３周的二步法培养，６周后收获细胞干、鲜重分别为１６．７２ｇ／Ｌ和１２７．４４ｇ／Ｌ，其生物量低于一步法，但总碱含量达３１．７６ｍｇ／ｇ．ｃｅｌｄｗ，培养基中生物碱量为２５．３１ｍｇ／Ｌ，每升培养液共可获生物碱５５６ｍｇ。二步法培养生物碱产率为一步法培养的１．７２倍。以上结果表明：二步培养法能够较好地解决黄连培养物生物量增长与次生代谢产物积累之间的矛盾，大大提高目的产物的产率，具有一定的实用价值。
In order to improve the unit yield of secondary metabolites produced by plant cell culture, the authors explored the production of berberine alkaloids by using two-step suspension culture of rhizomes. The results showed that: After one-step suspension culture of Coptis cells for 6 weeks, the dry and fresh weight of the cells were 20.96g / L and 174.92g / L, respectively. The weight gain was about 700%. The total alkaloid content in the cells was 14.79mg / G. celdw, the amount of alkaloids in the culture medium is 12.96mg / L, a total of 323mg of alkaloids per liter of culture solution can be harvested. The cells were cultured in a two-step method, which was cultured in the growth medium for 3 weeks and then in the synthetic medium for 3 weeks. After 6 weeks, the stems were harvested and the fresh weight was 16.72 g / L and 127.44 g / L, respectively The biomass was lower than the one-step method, but the total alkalinity content reached 31.76mg / g. celdw, the amount of alkaloids in the medium is 25.31mg / L, and a total of 556mg of alkaloid per liter of culture solution can be obtained. The yield of alkaloids by two-step method was 1.72 times of one-step culture. The above results show that the two-step culture method can solve the contradiction between the growth of the coptis culture and the accumulation of the secondary metabolites, and greatly improve the yield of the target product, which has a certain practical value.