不同培养条件对悬浮培养茶叶细胞生长及茶氨酸合成的影响*

婺源绿茶嫩叶用MS 培养基( 加IBA 2 mgPL, 6-BA 4 mgPL) 进行茶叶愈伤组织悬浮培养, 研究了不同培养条件对茶叶细胞悬浮培养过程中细胞生长与茶氨酸合成的影响。结果显示, NH4+PNO3- 110P6010 mmolPL、K+ 10010 mmolPL、Mg2+ 310mmolPL、H2PO4- 310 mmolPL、蔗糖3010 gPL、水解酪蛋白210 gPL 条件下, 茶叶细胞生长量和茶氨酸积累量均达到最高值; 提高培养基中蔗糖和水解酪蛋白浓度可使细胞对数生长期和稳定期得到延长, 从而有利于茶氨酸积累; H2 PO4- 浓度主要影响细胞生长速率和茶氨酸积累速率的同步性, 低H2 PO4- 浓度环境中茶氨酸积累速率峰值滞后于细胞增长速率峰值, 高H2PO4- 浓度环境中早于细胞生长速率峰值出现时间; K+ 和Mg2+ 对细胞生长的影响不明显, 但影响细胞茶氨酸合成酶活性, 维持适量的K+和Mg2+ 有利于茶氨酸积累。添加盐酸乙胺可大幅度提高茶氨酸积累量, 并且先加入一定量盐酸乙胺再每天进行少量补充, 茶氨酸合成量比一次性加入的效果要好。茶叶细胞生长和茶氨酸积累高峰期在整个培养过程的第19~ 22 天出现, 从生产效率考虑, 培养周期以19~ 22 天为宜。

Effects of Different Culture Conditions on Cell Growth and Theanine Biosynthesis in Suspension Cells of Camellia sinensis ( Theaceae)

The suspension culture of callus obtained from tender leave of Camellia sinensis was conducted withMS mediumplus IBA 2 mgPL and 6-BA 4 mgPL in a fermenter. The effects of different factors on the growth of tea cells and the amount
of synthesized theaninewere analyzed. The results showed that both the tea- cell growth and theanine accumulation reachedtheir peak period from 19 to 22 days after culture. Both growth of the cells and the amount of theanine reached optimal va-lues when the medium contained NH4+PNO3- 110P601 0 mmolPL, K+ 10010 mmolPL, Mg2+ 310 mmolPL, H2 PO4- 310mmolPL, sucrose 3010 gPL and protein 210 gPL. Both the logarithmic and stationary phases were enhanced when theamount of sucrose and protein were raised in the medium. Alteration of H2PO4- concentration in the medium had an effecton the simultaneity of the cel-l growth and theanine accumulation. The activity of theanine- synthatase ( L-glutamate: ethy-lamine lingase) can be affected by K+ andMg2+ . Accumulation of theanine could be greatly raised when ethy-l amine wassupplemented in the medium. The efficiency of ethy-l amine on the accumulation of theanine is related to the addition methodi1 e. sequential supplement of ethy-l amine into the medium was better than single addition of that.