光呼吸对碳源诱导雨生红球藻积累虾青素的影响

doi:10.13560/j.cnki.biotech.bull.1985.2025-0776

摘要/Abstract

摘要：

目的雨生红球藻是强抗氧化性虾青素产品的天然优质来源，补充碳源来提高雨生红球藻虾青素产量是一种有效策略。探究内源光呼吸途径对碳源诱导虾青素合成的影响，为开发高效虾青素生产工艺提供新思路。方法设计CK组（正常对照组，基础BG-11培养基）、A组（补充醋酸钠）和H组（补充碳酸氢钠）3组不同培养基，在未添加或添加光呼吸抑制剂CM的情况下，通过分析色素积累、生物量、总光合速率、OJIP荧光诱导曲线及其叶绿素荧光参数变化，解析光呼吸途径对碳源诱导雨生红球藻虾青素积累的影响。结果补充碳源醋酸钠或碳酸氢钠提高了雨生红球藻的干重和虾青素/叶绿素比值，其中醋酸钠显著促进了虾青素积累。然而使用CM抑制光呼吸后，干重、虾青素含量、虾青素/叶绿素比值显著降低。同时，补充碳源（A组、H组）导致总光合速率显著降低，光呼吸受阻使总光合速率进一步降低。此外，补充碳源或CM均使OJIP曲线的形状和荧光强度发生改变。在CK或A组条件下抑制光呼吸，对最大光化学效率（Φ_P0）和电子传递效率（Φ_E0）无影响，而单位面积内活性PSⅡ反应中心的数量（RC/CS₀）与单位活性反应中心吸收的光能（ABS/RC）、捕获的激发能（TR₀/RC）和用于电子传递的能量（ET₀/RC）均显著降低，A+CM组在J点的相对可变荧光强度（V_J）较A组显著增高。结论强光胁迫下添加醋酸钠再抑制光呼吸后，首先降低了PSII活性反应中心数量，继而损伤了PSII受体侧，导致光合作用光能吸收和能量利用受阻，从而进一步损伤雨生红球藻细胞，导致干重、虾青素含量和虾青素/叶绿素比值降低，证实光呼吸在有机碳源醋酸钠诱导雨生红球藻虾青素积累中发挥重要功能。

关键词: 雨生红球藻, 虾青素, 光呼吸, 醋酸钠, 叶绿素荧光

Abstract:

Objective Haematococcus pluvialis is the most promising natural source of the potent antioxidant astaxanthin. Carbon source supplementation is an effective strategy to enhance astaxanthin yield. Investigating the effect of endogenous photorespiration on astaxanthin accumulation induced by carbon sources would provide novel insights for optimizing production of H. pluvialis. Method Three different culture media were designed: Group CK: Normal control group, basic BG-11 medium. Group A: Supplemented with sodium acetate. Group H: Supplemented with sodium bicarbonate. The effects of photorespiration pathway on carbon source induced accumulation of astaxanthin in H. pluvialis were investigated by analyzing pigment accumulation, biomass, total photosynthetic rate, OJIP fluorescence induction curve, and chlorophyll fluorescence parameters, in the absence or presence of CM (photorespiration inhibitor) during the incubation, respectively. Result Supplementing with carbon sources such as sodium acetate or sodium bicarbonate enhanced algal dry weight and astaxanthin/chlorophyll ratio, with sodium acetate significantly increased astaxanthin accumulation. However, CM-mediated photorespiration inhibition markedly reduced the dry weight, astaxanthin content, and astaxanthin/chlorophyll ratio. Moreover, carbon supplementation (Group A and H) reduced total photosynthetic rate, further exacerbated by photorespiration inhibition induced by CM. Both carbon sources supplementation and CM altered OJIP curve profiles and fluorescence intensity. Under CK/A conditions, photorespiration inhibition unaffected maximum photochemical efficiency (Φ_P0) or electron transport efficiency (Φ_E0), but significantly decreased the amount of active PSII reaction centers (RC/CS₀), specific energy fluxes for absorption (ABS/RC), trapping flux of excitation energy (TR₀/RC), and electron transport flux (ET₀/RC) of per active reaction center. Compared with group A, Group A+CM presented notably higher relative variable fluorescence at J-step (V_J). Conclusion Under high-light stress, the addition of sodium acetate followed by inhibition of photorespiration first reduced the number of PSII active reaction centers, which then damaged the PSII receptor side, leading to obstruction of photosynthetic light energy absorption and utilization, and further damaging algal cells of H. pluvialis, resulting in reduced dry weight, astaxanthin content, and astaxanthin/chlorophyll ratio. This confirms that photorespiration plays an important role in the accumulation of astaxanthin in H. pluvialis induced by organic carbon source sodium acetate.

Key words: Haematococcus pluvialis, astaxanthin, photorespiration, sodium acetate, chlorophyll fluorescence

张春辉, 吉婧芳, 曹嘉敏, 马茜茜, 刘纹众, 季春丽, 张立涛, 李润植. 光呼吸对碳源诱导雨生红球藻积累虾青素的影响[J]. 生物技术通报, 2025, 41(10): 110-120.

ZHANG Chun-hui, JI Jing-fang, CAO Jia-min, MA Xi-xi, LIU Wen-zhong, JI Chun-li, ZHANG Li-tao, LI Run-zhi. Effect of Photorespiration on Astaxanthin Accumulation in Haematococcus pluvialis Induced by Carbon Sources[J]. Biotechnology Bulletin, 2025, 41(10): 110-120.

图/表 9

表1 各处理组培养基配方

Table 1 Medium formula for each treatment group

处理组 Treatment	基础培养基 Basal medium	外接碳源 Carbon source supplementation（1 g/L）	是否补充光呼吸抑制剂CM Absence or presence of photorespiration inhibitor CM（0.1 mmol/L）
CK	BG-11	无	否
CM	BG-11	无	是
A	BG-11	醋酸钠	否
A+CM	BG-11	醋酸钠	是
H	BG-11	碳酸氢钠	否
H+CM	BG-11	碳酸氢钠	是

图1 不同碳源培养下光呼吸抑制剂CM对雨生红球藻中虾青素含量和虾青素/叶绿素比值的影响a，c，e，g：虾青素含量；b，d，f，h：虾青素/叶绿素比值

Fig. 1 Effects of photorespiration inhibitor CM on the astaxanthin content and astaxanthin/chlorophyll ratio in H. pluvialis cultured with different carbon sourcesa, c, e, g: Astaxanthin content; b, d, f, h: Astaxanthin/chlorophyll ratio

图2 不同碳源培养6 d时光呼吸抑制剂CM对雨生红球藻干重（a）和总光合速率（b）的影响

Fig. 2 Effects of photorespiration inhibitor CM on dry weights (a) and total photosynthetic rates (b) in H. pluvialis cultured with different carbon sources

图3 不同碳源培养下光呼吸抑制剂CM对雨生红球藻瞬时叶绿素荧光曲线（OJIP）的影响

Fig. 3 Effects of photorespiration inhibitor CM on the chlorophyll a fluorescence (OJIP) transients in H. pluvialis cultured with different carbon sources

图4 不同碳源培养2 d和6 d时，光呼吸抑制剂CM对瞬时叶绿素荧光曲线（OJIP）的影响

Fig. 4 Effects of photorespiration inhibitor CM on the chlorophyll a fluorescence (OJIP) transients at 2 d and 6 d in H. pluvialis cultured with different carbon sources

图5 不同碳源培养下光呼吸抑制剂CM对雨生红球藻细胞相对可变荧光强度（VJ和WK）的影响a，c，e，g：相对可变荧光强度（VJ）；b，d，f，h：K点的相对可变荧光强度（WK）

Fig. 5 Effects of photorespiration inhibitor CM on relative variable fluorescence at J-step (VJ) and at K-step (WK) in H. pluvialis cultured with different carbon sourcesa, c, e, g: Relative variable fluorescence at J-step (VJ); b, d, f, h: Relative variable fluorescence at K-step (WK)

图6 不同碳源培养下光呼吸抑制剂CM对雨生红球藻最大光化学（ΦP0）和用于电子传递的量子产额（ΦE0）的影响a，c，e，g：最大光化学效率（ΦP0）；b，d，f，h：用于电子传递的量子产额（ΦE0）

Fig. 6 Effects of photorespiration inhibitor CM on the maximum photochemical efficiency (ΦP0) and the quantum yield of electron transport (ΦE0) in H. pluvialis cultured with different carbon sourcesa, c, e, g: The maximum photochemical efficiency (ΦP0); b, d, f, h: The quantum yield of electron transport (ΦE0)

图7 不同碳源培养下光呼吸抑制剂CM对雨生红球藻细胞单位面积内活性PSII反应中心的数量（RC/CS0）和单位活性反应中心吸收的光能（ABS/RC）的影响a，c，e，g：单位面积内活性PSII反应中心的数量（RC/CS0）；b，d，f，h：单位活性反应中心吸收的光能（ABS/RC）

Fig. 7 Effects of photorespiration inhibitor CM on the amount of active PSII reaction centers (RCs) per excited cross section (RC/CS0), and the specific energy fluxes (per RC) for absorption (ABS/RC) in H. pluvialis cultured with different carbon sourcesa, c, e, g: The amount of active PSII reaction centers (RCs) per excited cross section (RC/CS0); b, d, f, h: The specific energy fluxes (per RC) for absorption (ABS/RC)

图8 不同碳源培养下光呼吸抑制剂CM对雨生红球藻单位活性反应中心捕获的激发能（TR0/RC）和单位活性反应中心捕获的用于电子传递的能量（ET0/RC）的影响a，c，e，g：单位活性反应中心捕获的激发能（TR0/RC）；b，d，f，h：单位活性反应中心捕获的用于电子传递的能量（ET0/RC）

Fig. 8 Effects of photorespiration inhibitor CM on the trapping flux of excitation energy per active reaction center (TR0/RC), and the electron transport flux per active reaction center (ET0/RC) in H. pluvialis cultured with different carbon sourcesa, c, e, g: The trapping flux of excitation energy per active reaction center (TR0/RC); b, d, f, h: The electron transport flux per active reaction center (ET0/RC)

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