碳氮磷营养对百蕊草悬浮培养细胞生长的影响.

[ Objective] A good suspension cell culture system of Thesium chinese Turcz. is established, so as to lay a foundation for the subsequent use of genetic engineering to transform the metabolic pathway of Thesium chinese Turcz., obtain new varieties of high-order metabolites, and realize the substitution of wild resources. [ Method] On the basis of exploring the law of growth and nutrient consumption of Thesium chinese Turcz. suspension cells, the initial input of liquid nitrogen and phosphorus and the ratio of ammonium nitrogen to nitrate nitrogen (NH+4/ NO-3 ) were optimized. [Result] In the 30 days culture process, the growth curve of suspension cells showed an obvious delay period ( 0 - 6 days), logarithmic growth period ( 7 - 15 days) ), stable growth period ( 16 - 24 days) and decay period ( after 24 days) . The results showed that the dry matter mass of suspension cells reached the maximum when the initial total nitrogen concentration was 70 mmol/L and the phosphate concentration was 1. 3 mmol/L for 28 days, and it was unhealthy for the growth of cells when the nitrogen concentration and phosphate were higher and lower. On this basis, the demand of suspension cultured cells for nitrate nitrogen and ammonium nitrogen were explored. The results showed that the suspension cells during the culture process consumed more nitrate nitrogen than ammonium nitrogen, while the utilization and consumption of ammonium nitrogen precedes that of nitrate nitrogen and too high concentration of ammonium nitrogen would inhibit cell growth. It was most suitable for cell growth when NH+4 / NO+3- was 20/50. [ Conclusion] The growth curve of Thesium chinese Turcz. suspension cells shows ' S ' as taking MS as the basic medium, the concentration of nitrogen is 70 mmol/L, the concentration of phosphate is 1. 3 mmol/L, and the concentration of NH+4/NO-3 is 20/50, which is more conducive to the suspension culture of Thesium chinese cells. Under these conditions, the consumption of total sugar, sucrose, reducing sugar, nitrate nitrogen and inorganic phosphorus are faster in the logarithmic growth period, and the consumption of ammonium nitrogen is faster than nitrate nitrogen in the early growth period. [ABSTRACT FROM AUTHOR]