Effects of Light Exposure on Cell Culture and Corilagin Synthesis of Longan
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摘要:
目的 研究不同光照条件对龙眼胚性悬浮细胞培养及柯里拉京代谢合成的影响，以期为龙眼胚性悬浮细胞大规模培养、工业化生产柯里拉京奠定理论基础。 方法 基于龙眼胚性悬浮细胞体系的建立，采用5 L新型搅拌式生物反应器放大培养龙眼胚性悬浮细胞，探讨黑暗和光照处理对龙眼胚性悬浮细胞生长、柯里拉京含量、蔗糖消耗以及培养体系中pH和DO（溶氧）的动态变化规律。 结果 龙眼胚性悬浮细胞在黑暗和光照条件下其生长动态大致均呈“S”形曲线，最大增重分别为4.74 g·L-1和3.71 g·L-1；黑暗培养到第7 d龙眼胚性悬浮细胞体内柯里拉京含量达到最大值8.72 mg·g-1，是光照条件下的2.8倍，但在细胞培养液中柯里拉京含量明显低于光照条件（P < 0.05）。光照加快了龙眼胚性悬浮细胞对蔗糖的消耗速度，但对培养体系中的pH和DO无明显影响。 结论 生物反应器大规模培养过程中，黑暗条件利于龙眼胚性悬浮细胞生长及细胞内柯里拉京含量的积累，蔗糖消耗、pH和DO是龙眼胚性悬浮细胞生长及柯里拉京合成的重要调控因子。 Abstract:Objective Effects of light exposure on embryogenic cell and corilagin formations of longan (Dimocarpus longan Lour.) in a suspension culture were investigated to provide theoretical reference for the large-scale operation of longan embryogenic cell culture and industrial production of corilagin. Method A 5 L stirring bioreactor was used in the experimentation. Dynamic changes on the cell growth, corilagin content, substrate consumption, pH, and dissolved oxygen (DO) in the medium under dark or light condition were monitored. Result The cellular growth curve showed an S shape with a peaked weight of 4.74 g·L-1 in the dark and 3.71 g·L-1 under the light. Cultured 7 days in the dark, the corilagin content in the longan cells reached a maximum of 8.72 mg·g-1, which was 2.8 times higher than that with the light exposure. But the corilagin in the medium was significantly less without than with the light exposure (P < 0.05). Light accelerated the sucrose consumption by the cells but had no significant effect on pH or DO in the medium. Conclusion For large-scale operations, the absence of light in an enclosed reactor would facilitate the embryogenic growth and corilagin accumulation. Meanwhile, process controls on sucrose, pH and DO were also important as these conditions regulated the cell growth and corilagin formation as well. -
Key words:
- Dimocarpus longan Lour. /
- cell culture /
- bioreactor /
- light exposure /
- corilagin
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图 3 龙眼胚性悬浮细胞培养液中蔗糖消耗量的变化
注：蔗糖消耗量(g·L-1)=20-培养液中蔗糖含量。
Figure 3. Sucrose consumption in medium during longan embryonic cell culture
Note:Sucrose consumption(g·L-1)=20-sucrose content in culture medium.
表 1 生物反应器放大培养龙眼胚性悬浮细胞相关参数
Table 1. Process conditions of longan embryonic cells suspension culture in bioreactor
培养参数
Parameters参数值
Value温度Temperature 25±2℃ 初始pH Initial pH 5.8 搅拌转速Speed of stirring 120 r·min-1 接种密度Inoculum density 40 g·L-1FW 通气量Ventilation 100 cm3·min-1 初始液位Initial level 4 L 培养基Medium MS+20 g·L-1蔗糖+1.0 mg·L-1
2, 4-D+100 mg·L-1肌醇培养天数Cultured day 9 d 光照强度(白光)Intensity of white light 32 μmol·m-2·s-1 光照周期(白光)Photoperiod of white light 12 h 
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表 2 龙眼胚性悬浮细胞培养液中柯里拉京的含量变化
Table 2. Changes on corilagin content in suspension medium for longan embryonic cell culture
培养天数
Cultured day/d柯里拉京含量Corilagin content/(mg·mL-1) 黑暗Dark 光照Light 1 ND ND 2 ND ND 3 ND (0.56±0.03)aA 4 (0.15±0.03)bB (0.84±0.05)aA 5 (0.17±0.04)bB (0.87±0.05)aA 6 (0.32±0.02)bA (0.44±0.05)aA 7 ND (0.27±0.03)aA 8 ND (0.18±0.03)aA 9 ND ND 注：ND代表UPLC未检测出细胞培养液中柯里拉京含量；不同大、小写字母分别表示差异极显著(P < 0.01)或差异显著(P < 0.05)。
Note：ND indicates that corilagin was not detected in culture medium by UPLC. Different uppercase letters represent highly significant difference at P < 0.01, and lowercase letters significant difference at P < 0.05.
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