EFFECTS OF EUCALYPTOL AND LIMONENE ON THE PHOTOSYNTHETIC ABILITIES IN CHLAMYDOMONAS REINHARDTII
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摘要: 为了揭示蓝藻挥发性有机化合物（VOCs）中两种主要萜烯类化合物桉树脑和柠檬烯对其他藻类的化感作用，研究了此两种化合物对莱茵衣藻细胞生长、光合素吸收光谱和光合性能的影响。结果表明，莱茵衣藻在1.6和4 mmol/L桉树脑以及0.4和0.8 mmol/L柠檬烯处理24h后，细胞生长受到明显抑制，与对照相比，细胞密度分别降低了16.7%、50.6%、29.1%和44.4%。同时，1.6和4 mmol/L柠檬烯处理会诱导藻细胞全部死亡。此外，莱茵衣藻光合色素在413、433、457和663 nm处的吸收峰均明显降低，光合色素的各成分发生明显降解，叶黄素甚至在0.8 mmol/L柠檬烯处理下完全降解消失。在桉树脑和柠檬烯处理后，莱茵衣藻从O点到P点的荧光强度均低于对照，并且随处理浓度升高而降低。同时，荧光诱导动力学参数中，φPo、Ψo、φEo、RC/CSMS、ABS/CSM、TRo/CSM、ETo/CSM和PIABS均明显降低，而DIo/CSM则明显升高，这表明桉树脑和柠檬烯可抑制莱茵衣藻PSⅡ量子产生和电子传递，并使吸收的光能以热的形式进行耗散。由此可见，桉树脑和柠檬烯可能通过引起其他藻细胞光合色素降解、降低光合性能而发挥化感作用。Abstract: This study investigated the allelopathic effects of eucalyptol and limonene, two main terpenoids from cyanobacteria VOCs, on other algae by focusing on cell growth, absorption spectra of photosynthetic pigments and photosynthetic abilities using Chlamydomonas reinhardtii. Eucalyptol at 1.6 and 4 mmol/L as well as limonene at 0.4 and 0.8 mmol/L remarkably inhibited C. reinhardtii cell growth by 16.7%, 50.6%, 29.1% and 44.4%, respectively. Moreover, limonene at 1.6 and 4 mmol/L killed the cells. In addition, an obvious reduction was found in the absorption peaks at 413, 433, 457 and 663 nm. The components of photosynthetic pigments were degraded markedly, and xanthophyll even disappeared by 0.8 mmol/L limonene treatment. When C. reinhardtii cells were treated by eucalyptol and limonene, the fluorescence intensity from O to P was lower compared to the control, and it was declined with the increased compound concentration. In chlorophyll fluorescence transient parameters, φPo, Ψo, φEo, RC/CSM, ABS/CSM, TRo/CSM, ETo/CSMand PIABS were declined remarkably, while DIo/CSM was raised remarkably, indicating that eucalyptol and limonene can inhibit the quantum production and electron transport in PSⅡ, and promote the absorbed solar energy to dissipate as heat. Therefore, eucalyptol and limonene might play allelopathic effects in cyanobacteria VOCs via inducing degradation of photosynthetic pigments and reducing photosynthetic abilities in other algae.
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Keywords:
- Allelopathy /
- Eucalyptol /
- Limonene /
- Absorption spectrum /
- Photosynthetic ability
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图 1 桉树脑和柠檬烯对莱茵衣藻细胞生长的影响
A. 桉树脑; B. 柠檬烯; CK. 对照; 下同
Figure 1. Effects of eucalyptol and limonene on the growth of C. reinhardtii cells
A. Eucalyptol; B. Limonene; CK. The control; the same applies below
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图 2 桉树脑和柠檬烯对叶绿素吸收光谱的影响
Figure 2. Effects of eucalyptol and limonene on chlorophyll absorption spectra
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图 3 桉树脑和柠檬烯对叶绿素4阶导数光谱的影响
419. 二乙烯基脱镁Chl. a; 438. 二乙烯基Chl. a; 468. 二乙烯基Chl. b; 475. 叶黄素; 623. 单乙烯基原叶绿素酸酯a; 645. 单乙烯基Chl. b; 663. 单乙烯基Chl. a
Figure 3. Effects of eucalyptol and limonene on 4th derivative of absorption spectra
419. Divinyl-pheophytin a, 438. Divinyl-chlorophyll a, 468. Divinyl-chlorophyll b, 475. Xanthophyll, 623. Monovinyl-protochlorophyllide a, 645. Monovinyl-chlorophyll b, 663. Monovinyl-chlorophyll a
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图 4 桉树脑和柠檬烯对叶绿素荧光诱导动力学曲线的影响
Figure 4. Effects of eucalyptol and limonene on chlorophyll fluorescence kinetics
表 1 桉树脑对莱茵衣藻荧光动力学参数的影响
Table 1 Effects of eucalyptol on chlorophyll fluorescence transient parameters in C. reinhardtii
参数Parameter CK 0.4 mmol/L 0.8 mmol/L 1.6 mmol/L 4 mmol/L φPo 0.76±0.01 0.75±0.01 0.72±0.02 0.71±0.03* 0.68±0.03* Ψo 0.49±0.01 0.47±0.02 0.48±0.03 0.46±0.01* 0.41±0.02** φEo 0.38±0.01 0.36±0.02 0.38±0.02 0.33±0.02** 0.31±0.01** ABS/CSM 76.03±5.97 71.90±6.60 69.73±2.82* 60.40±1.15** 50.65±8.90** TRo/CSM 57.92±4.17 55.41±2.26 53.30±1.98* 43.90±2.90** 35.53±7.93** ETo/CSM 6.78±0.64 6.74±0.07 5.94±0.38* 5.76±0.43** 4.95±0.58** DIo/CSM 17.17±1.89 17.45±0.84 19.37±0.83* 21.14±1.30** 25.81±1.34** RC/CSM 23.06±1.59 22.35±3.21 21.67±1.42 18.32±1.32** 13.65±3.24** PIABS 0.94±0.07 0.90±0.10 0.87±0.07 0.66±0.10** 0.61±0.12** 注: CK. 对照; *. 在24h时与对照相比, 差异在P＜0.05水平上显著; **. 在24h时与对照相比, 差异在P＜0.01水平上显著; 下同 Note: CK. The control; *. Compared to the control after 24h, significant difference at P ＜ 0.05 level; **. Compared to the control after 24h, significant difference at P ＜ 0.01 level; The same applies below 
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表 2 柠檬烯对莱茵衣藻荧光动力学参数的影响
Table 2 Effects of limonene on chlorophyll fluorescence transient parameters in C. reinhardtii
参数Parameter CK 0.2 mmol/L 0.4 mmol/L 0.8 mmol/L φPo 0.76±0.01 0.72±0.03 0.64±0.05** 0.33±0.07** Ψo 0.49±0.01 0.48±0.02 0.40±0.02** 0.31±0.01** φEo 0.38±0.01 0.36±0.02 0.30±0.02** 0.20±0.01** ABS/CSM 76.03±5.97 70.21±4.24* 63.50±8.56** 20.40±1.90** TRo/CSM 57.92±4.17 52.97±2.80* 45.09±3.14** 11.87±1.25** ETo/CSM 6.78±0.64 6.02±0.30* 5.96±0.68** 2.34±0.29** DIo/CSM 17.17±1.89 19.93±1.45* 24.35±2.86** 29.67±1.12** RC/CSM 23.06±1.59 21.81±1.44 15.94±4.67** 4.35±0.74** PIABS 0.94±0.07 0.84±0.08* 0.37±0.10** 0.27±0.03**
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