Callus Induction,stem Cell Culture And Follow-up Study Of Taxus

This paper is a systematic research on dedifferentiate callus and stem cells of Taxus.A regulatory strategy to the Taxane titer near industrialization level was primarily explored.A simpler assay method for high yield level was developed.The plant stem cell of Taxus was primarily identified in morphology.These above lay a good foundation for the sequential high-yield and stable-production of the large-scale plant cell culture.The main results are list below:The calli with the fastest growth was derived from two years stem section.The optimal formula of induction medium was the same as the one of subculture medium,both for:B5 medium basic salts?sucrose 30g/L?L-glutamine 300 mg/L?2,4-D 1.3 mg/L?6-BA 0.13mg/L?ascorbic acid 0.1 g/L Vc?agar 8 g/L.The optimal formula of suspension-cultured medium had been achieved via the orthogonal experiment on plant hormones,that could be described,B5 medium basic salts?sucrose 30g/L?L-glutamine 300 mg/L?2,4-D 1.3mg/L?6-BA 0.13 mg/L?ascorbic acid 0.1 g/L Vc.The established growth dynamics model showed that cell grew best when the subculture duration was set at the value of 14 days.The effects of different elicitors for secondary metabolite production in Taxus cell suspension were explored to find out the key factors.Then the dynamics model of secondary metabolite production of suspension-cultured cells was established.The data showed that the key factors were the state of seed,the ingredient in production medium and the main modulator added.The maximum titer of 10-DAB?Baccatin ??Taxol are 1.33mg/g?3.42mg/g?2.089mg/g,respectively.The study on stem cell of Taxus showed there is no diffrerence among the species Taxus cuspidata,taxus chinensis var.mairei and taxus madia.All could be induced a fine stem cell line,but the best explant is the peeled epidermis of stem of 3-5 years old.It was found that there is many and more vesicles and no large mature vacuole in stem cell with contrast of that in the dedifferentiated callus.