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为研究银杏细胞的耐盐能力和提高细胞培养生产黄酮的产量,通过添加0、50、100、150 mmol/L的NaCl到液体培养基中进行悬浮细胞振荡培养,对盐处理下银杏悬浮细胞的生长、耐盐能力和黄酮积累的动态变化进行了研究。结果表明:在盐处理早期(4~8 d),各处理的细胞生长量随着盐处理浓度的增加而增加,而在后期(12~20 d)则随着盐处理浓度的增加而下降。中、高浓度盐处理(100~150 mmol/L)的细胞在培养早期表现出对盐刺激有一定的耐受性;8~16 d时盐胁迫加强,细胞通过提高脯氨酸含量增强渗透调节的能力,同时通过促进细胞内黄酮的积累增强抗氧化能力,减少盐胁迫对细胞的伤害;而在20 d时盐胁迫强度超过细胞的自我调节能力,细胞受到严重的伤害而出现褐化。50 mmol/L NaCl处理的悬浮细胞生长受抑制较轻,PAL活性在第8天最高,高的黄酮含量和产量(129.0 mg/L)出现在第12天,在整个处理期内能够维持生长。100 mmol/L盐处理的细胞在第16天时黄酮含量比对照增加了29.0%,但总黄酮产量(98.4 mg/L)只比对照增加了10.9%。150 mmol/L盐处理的细胞在第8天和第16天都维持较高水平的黄酮含量,第8天时比对照增加了66.3%,总黄酮产量最高达104.0mg/L。因此50 mmol/L和150 mmol/L NaCl处理下,悬浮培养的银杏细胞分别在第12天和第8天时收获较好。
In order to study salt tolerance of Ginkgo biloba cells and increase the yield of flavonoids produced by cell culture, suspension culture was carried out by adding 0, 50, 100 and 150 mmol / L NaCl to liquid medium. Growth, salt tolerance and accumulation of flavonoids were studied. The results showed that in the early salt treatment (4 ~ 8 d), the cell growth of each treatment increased with the increase of salt concentration, but decreased with the increase of salt concentration in the later period (12 ~ 20 d). Medium and high concentrations of salt treatment (100 ~ 150 mmol / L) of the cells in the early culture showed salt tolerance to a certain extent; 8 to 16 d when the salt stress to strengthen the cells by increasing the proline content and enhance osmotic adjustment At the same time, by increasing the accumulation of flavonoids in cells, the antioxidative capacity was enhanced and the damage to cells by salt stress was reduced. At 20 days, the salt stress exceeded the self-regulation ability of cells, and the cells were severely damaged and browned. The growth of suspension cells treated with 50 mmol / L NaCl was inhibited to a minimum, and PAL activity was the highest on the 8th day. The high flavonoid content and yield (129.0 mg / L) appeared on the 12th day and maintained the growth during the whole treatment period. Flavone content of 100 mmol / L salt-treated cells increased by 29.0% compared with the control on the 16th day, but the total flavonoid yield (98.4 mg / L) increased by only 10.9% compared with the control. 150 mmol / L salt-treated cells maintained high levels of flavonoids on the 8th day and the 16th day, increased 66.3% on the 8th day and the highest total flavonoid yield reached 104.0mg / L. Therefore, under the treatment of 50 mmol / L and 150 mmol / L NaCl, the ginkgo cells cultured in suspension were better harvested on the 12th and the 8th day respectively.