葡萄悬浮细胞长期培养过程中不同继代条件下花青素生产的不稳定性

Instability of anthocyanin composition under different subculture conditions during long-term suspension cultures of Vitis vinifera L. var. Gamay Fréaux

摘要:

为了深入研究植物细胞培养生产次生代谢产物不稳定性的机制，以葡萄细胞作为模式体系，研究悬浮培养过程中花青素合成的不稳定性。除了用常规的花青素总含量来表征花青素的生物合成之外，还采用HPLC测定花青素不同组分的含量。结果表明，在长期的继代培养过程中，不仅花青素的含量而且花青素的组成也表现出明显的不稳定性。首次采用了不稳定系数 (δ) 和因素得分 (Factor scores) 来表征植物细胞培养过程中次生代谢生产的不稳定性。培养条件对花青素生物合成的影响实验结果表明，继代周期和接种量均能诱发次生代谢的不稳定性表达，其中接种量的影响相对更大。在考察的 (6.5 d，2.00 g)，(7 d，2.00 g)，(7.5 d，2.00 g)，(7 d，1.60 g) 和 (7 d，2.40 g) 五种不同的继代周期和接种量组合条件中，7 d继代周期和1.60 g接种量最有利于保持花青素的稳定生产。

Abstract:

The instability of secondary metabolite production is a ubiquitous problem in plant cell culture. In order to understand the instability in plant cell culture, we investigated anthocyanin accumulation in suspension cultures of Vitis vinifera, as a model system, in our laboratory. Not only the anthocyanin contents but also its composition exhibited instability along with the long-term subculture. New methods were developed to indicate the instability of plant cell culture. Both the definition of instability coefficient (δ) and the application of factor scores were the first time in this field. To examine the effects of culture conditions on instability of anthocyanin biosynthesis, different subculture cycles and inoculum sizes had been investigated. Subculture cycle and inoculum size were both environmental cues driving the instability. Compared with subculture cycle, inoculum size was more effective in working on the instability of anthocyanin accumulation. Among all the conditions investigated in our study, (6.5 d, 2.00 g), (7 d, 2.00 g), (7.5 d, 2.00 g), (7 d, 1.60 g) and (7 d, 2.40 g), the condition of 7 d-subculture cycle together with 1.60 g-inoculum size was the best one to keep the stable production of anthocyanins.