万寿菊悬浮培养细胞生产游离叶黄素工艺的研究

Study on the Process of Producing Free Lutein by Cell Suspension Cultures of Tagetes erecta L.

建立万寿菊悬浮培养细胞生产游离叶黄素的方法。以万寿菊花瓣为外植体构建得到能够生产游离叶黄素的悬浮培养细胞系。研究细胞培养时间、光照和黑暗培养以及诱导子茉莉酸甲酯、水杨酸对细胞中游离叶黄素合成的影响,建立万寿菊悬浮培养细胞生产游离叶黄素的工艺。结果表明,万寿菊悬浮培养细胞中游离叶黄素的合成和细胞生物量的积累显示出相同的变化规律。在培养的第14 d,进入稳定期,细胞中游离叶黄素产率为1.29±0.09 mg/L,为第0天的8.48倍。培养14 d,光照培养细胞中游离叶黄素产率为暗培养细胞的5.7倍。分别添加茉莉酸甲酯和水杨酸对培养细胞生产游离叶黄素均无显著性影响。上述结果说明,细胞生产游离叶黄素与细胞生物量积累是偶联的。细胞最佳收获时间为生长的第14 d。光照能够显著促进细胞生产游离叶黄素。茉莉酸甲酯和水杨酸不能促进细胞中游离叶黄素的合成。

A method for producing free lutein by cell suspension cultures of Tagetes erecta L.was established.Suspension cultures of a free lutein-producing cell line derived from marigold petals.To establish process for the production of free lutein by cell suspension cultures of Tagetes erecta L.,the effects of culture time,light and dark periods,and elicitors including methyl jasmonate and salicylic acid on the synthesis of free lutein in cells were studied.The results showed that the pattern of change in free lutein synthesis and in biomass accumulation were similar.On the 14th day,the cultures entered stationary phase and the yield of free lutein in the cells was 1.29±0.09 mg/L,which was 8.48 times that of the 0th day.On the 14th day,the yield of free lutein in light-cultured cells was 5.7 times that of dark-cultured cells.The addition of methyl jasmonate and salicylic acid respectively had no significant effect on the production of free lutein by cultured cells.The results indicated that free lutein production was coupled to biomass production.The optimal time for cell harvest was the 14th day after inoculation.Light significantly enhanced free lutein production in cells.Methyl jasmonate and salicylic acid did not promote free lutein synthesis in cells.