红光下培养方式对雨生红球藻细胞增殖及其活力的影响

EFFECTS OF CULTURE METHOD ON CELL PROLIFERATION AND CELL ACTIVITY OF HAEMATOCOCCUS PLUVIALIS GROWING UNDER RED LIGHT

  • 摘要: 研究重点针对雨生红球藻绿色游动细胞的增殖培养阶段,分析了在利于细胞增殖的红光条件下,几种培养方式的调整对增殖过程和细胞活力的影响。结果显示:(1)在红光下,增殖平台期维持时间长,细胞活力稳定,细胞中性脂无累积,但进入平台期前,细胞中性脂有规律波动,进入平台期后相对稳定;通过更新率为20%的半连续培养,细胞数产出较批次培养提高57%;半连续培养中细胞呈现胁迫调节的时间较批次培养晚。随着培养时间增加,半连续培养下细胞营养盐吸收能力降低。(2)初始接种密度与细胞增殖速率及细胞光合活力呈负相关:初始密度低的细胞增殖速率较高,细胞光合作用活力高。(3)在培养过程中添加CO2时,最大密度均有提高,达6.0105 cells/mL,较无添加组提高54%;细胞分裂速率均有提高,但红光下较白光下增殖速率高(分别为0.223/d和0.198/d);添加CO2降低培养液pH,利于维持适宜增殖的pH环境。叶绿素荧光参数以及细胞粒径在红光和白光下有显著差异:红光下,Fv/Fm显著高于白光下;红光下补充CO2显著减小细胞粒径,而白光下粒径无显著变化。研究结果显示,在红光下,采用间断式半连续培养补充CO2培养绿色游动细胞,有利于提升细胞活力与产出。

     

    Abstract: The current study investigated methods culturing green motile cells of Haematococcus pluvialis under red light on cell density, cell growth rate and cell activity. In red light, the stationary growth phase in batch culture maintained for long time, and the cell photosynthetic activity maintained stable high. There was no neutral lipid accumulation in the cell growth in red light. Biomass yields from a semi-continuous culture of 20% medium renew rate was 57% higher than those from the batch culture. The variation of nutrient absorption rate and chlorophyll fluorescence parameter showed that cellular stress regulation started earlier in batch culture than those in semi-continuous cultures. The cell growth rate and photosynthetic activity were negatively related to the initial cell density in a batch culture. Culture of lower initial density had higher growth rate and higher photosynthetic activity. The effects of CO2 on the cultures under red light or under white light were conditionally dependent. Adding CO2 increased maximum biomass by 54%. The growth rates increased as while, however, that in the red light was higher (0.223/d) than that in white light (0.198/d). The pH values were not related with the light quality under both red light and white light, and pH decreased to the suitable level of pH 78. Chlorophyll parameter Fv/Fm significantly increased by red light with CO2 addition while it is not mediated by white light. Cell size significantly decreased by red light with CO2 addition while it was no regulated by white light. These results revealed that the method of intermittent semi-continuous culture for green motile cells of H. pluvialis had higher cell yield with high activity, which is a plausible strategy in astanxanthin industry.

     

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