Abstract

The photosynthetic process of phytoplankton is the basis of the material circulation and energy flow of the ecosystem. The rapid and accurate measurement of phytoplankton photosynthesis rate is of great significance to water ecological environment monitoring, marine resource assessment and global climate change prediction. On the basis of “Bio-Optical” model, a photosynthetic rate measurement method based on tunable pulsed light induced fluorescence kinetics was put forward in this paper. The chlorophyll fluorescence was used as the probe of photosynthesis process, and the phytoplankton photosynthetic rate was evaluated by the photosynthetic electron transport rate. Comparative experiment results showed that the photosynthetic electron transport rate measured by fluorescence kinetic method under different conditions of DCMU, culture light and nutrients (nitrogen) were consistent with the photosynthetic oxygen evolution rate measured by oxygen evolution method, and the correlation coefficient R2 were 0.934, 0.957 and 0.955 respectively.

© 2018 Optical Society of America under the terms of the OSA Open Access Publishing Agreement

1. Introduction

As a primary producer in water, phytoplankton possess less than 1% of the total vegetation biomass globally, but its productivity accounts for about 45% of total plant productivity and 40% of global total carbon sequestration [1]. The phytoplankton photosynthesis is the basis of the material circulation and ecosystem energy flow, and affects the global biogeochemical cycle and climate change fundamentally. The rapid and accurate measurement of phytoplankton photosynthetic rate is of great significance to water ecological environment monitoring, marine resource assessment and global climate change prediction.

In the photosynthesis process, the photosynthetic tissues are driven by light produce oxygen and electrons by cracking water molecules. The electron transmitted by reaction center and multistep electron acceptors finally generate reduction force and participate in photosynthetic carbon sequestration. In the black and white bottle method, 14C tracer method and other traditional gas exchange methods, the photosynthetic rate is calculated by measuring the photosynthetic oxygen evolution rate or the photosynthetic carbon fixation rate, which require “sampling-incubation-offline analysis”. These methods have the disadvantages of time-consuming, low efficiency and complicated procedure, and cannot meet the rapid observation demand in ecological and environmental monitoring. In 1992, Falkowski et al. [2] first proposed the photosynthesis analysis method using chlorophyll fluorescence kinetics. The method employs the fluorescence of reaction center as the probe to analyze the light absorption, transmission and conversion efficiency of cells and the phytoplankton photosynthetic capacity can be obtained directly and rapidly. This method possesses the advantages of rapid measurement, non-pretreatment, non-pollution and non-invasion, and thus it’s an advanced tool for the rapid sensing of phytoplankton photosynthesis in natural conditions. However, the chlorophyll fluorescence kinetics is affected by many photosynthetic chemical processes, leading to a complex theoretical model. Thus there would be large errors if the complex theoretical model is directly used to fit the fluorescence kinetics curve. In recent decades, in order to improve the measurement accuracy of photosynthetic parameters, Schreiber and Kolber et al. [3,4] put forward Pulse Amplitude Modulation (PAM), Fast Repetition Rate (FRR) and some other kinds of photosynthesis analysis techniques using chlorophyll fluorescence successively. However, these techniques are mainly used to measure the local photosynthetic parameters and cannot measure the photosynthetic rate.

In this paper, a photosynthetic rate measurement method based on Tunable Pulsed Light Induced Fluorescence kinetics (TPLIF) was proposed. The phytoplankton photosynthetic rate was evaluated by measuring photosynthetic electron transport rate. The validity of the fluorescence kinetics method was verified by comparing with the photosynthetic oxygen evolution method.

2. Principle and method

The energy flow of phytoplankton photosynthesis is shown in Fig. 1. Driven by light energy, the water molecules are cracked and electrons are produced. The electrons transmitted by reaction center and multistep electron acceptors finally generate reduction force and participate in Calvin cycle photosynthetic carbon sequestration. From the perspective of energy transport, photosynthetic rate is essentially photosynthetic electron transport rate Pe. According to “Bio-Optical” model [5], photosynthetic electron transport rate depends on the ambient light intensity E, light energy absorption efficiency of photosystem II (PSII) APSII, efficiency of charge separation and conversion Te, as defined in Eq. (1).

 figure: Fig. 1

Fig. 1 Diagram of the energy flow in phytoplankton photosynthesis (modified from [5]).

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Pe=E×APSII×Te

In order to accurately obtain the two key elements of photosynthetic electron transfer rate Pe, light energy absorption efficiency of photosystem II (PSII) APSII and efficiency of charge separation and conversion Te, a photosynthetic rate measurement method based on TPLIF was studied. High light pulse is employed to stimulate the dark-adapted phytoplankton cells, and during the QA single-turnover period (photochemical reaction inactivated), the photosynthetic electron transport chain is blocked by a large number of electrons produced by photosynthetic reaction center. As a consequence, the fluorescence increases rapidly from the minimum fluorescence Fo to the maximum fluorescence Fm, that is the fast phase fluorescence. The high light pulse is then turned off and the photochemical reaction process is activated. The fluorescence decays exponentially, and the decrease of the fluorescence, that is the relaxation fluorescence, is recorded using weak pulse light, as shown in Fig. 2.

 figure: Fig. 2

Fig. 2 Measurement process of tunable pulsed light induced fluorescence kinetics.

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Based on the photosynthesis energy flow theory, the fast phase fluorescence F(t) and relaxation fluorescence G(t) can be formulated by Eqs. (2) and (3) respectively. From these two equations, minimum fluorescence Fo, maximum fluorescence Fm, functional absorption cross section σPSII, QA average reoxidation time τQA and other photosynthetic fluorescence parameters that dominate the process of photosynthesis can be inverted [6].

F(t)=Fo+(FmFo)(1exp(σPSII0tEdt))
G(t)=Fo+(FmFo)(exp(t/τQA))

According to “Bio-Optical” model, the light energy absorption efficiency of PSII APSII depends on functional absorption cross section σPSII and light-harvesting efficiency ΦT. The efficiency of charge separation and conversion Te depends on light-harvesting efficiency ΦT, photochemical quenching ΦP and PQ average reduction rate ΦQA, as shown in Eqs. (4) and (5). Photochemical quenching ΦP and PQ average reduction efficiency ΦQA can be evaluated by photosynthetic fluorescence parameters, as shown in Eqs. (6) and (7), where Fm' and Fo' are maximum fluorescence and minimum fluorescence under light adaptation respectively.

APSII=σPSII/ΦQA
Te=ΦT×ΦP×ΦQA
ΦP=(FmFo)/(Fm'Fo')
ΦQA=1/τQA

Combining Eqs. (4)–(7) to Eq. (1), a photosynthetic rate analysis model based on TPLIF is established. The photosynthetic rate evaluated by photosynthetic electron transport rate Pe can be calculated in Eq. (8).

Pe=E×APSII×Te=E×σPSII×(FmFo)/(Fm'Fo')×1/τQA

3. Apparatus and experiment

For evaluating photosynthetic rate, the photosynthetic electron transport rate Pe, photosynthetic oxygen evolution rate PO2 and photosynthetic carbon fixation rate PCO2 are in consistent [7]. In this paper, the photosynthetic electron transport rate Pe measured by TPLIF is compared with the photosynthetic oxygen evolution rate PO2 measured by liquid-phase oxygen measurement system (Hansatec, Chlorolab2) to verify the validity of the fluorescence kinetics measurement method of photosynthetic rate.

3.1 TPLIF measurement system

The system consists of four parts: excitation-emission optical system, tunable fast-pulsed light source, fluorescence detection module and computer control system, as shown in Fig. 3. The excitation-emission optical system adopts orthogonal optical path structure and is combined with the filter system, the interference of excitation light to the fluorescence signal is deeply inhibited. The tunable fast-pulsed light source is composed of LEDs array with a wavelength of 465nm. The excitation intensity is accurately regulated by changing power supply voltage, frequency and duty ratio of driving pulse. A saturated intensity that is higher than 30000 μmol quanta/m2/s (hereinafter referred μmol quanta/m2/s to as L) is used to regulate photosynthetic electron transfer process, and a weak intensity of several L is employed to record the fluorescence change process [8]. Photomultiplier tube (Hamamatsu, h6779-01) is used as photoelectric detector in the fluorescence detection module. The fast phase fluorescence signal is acquired by high speed AD without distortion following I/V conversion, variable gain and low pass filtering. The relaxation fluorescence signal is sensitively detected by integral amplification and high resolution AD. The computer control system is mainly used for the synchronous control of pulsed light excitation and fluorescence detection, as well as the collection of fluorescence kinetic curves and calculation of photosynthetic electron transport rate Pe.

 figure: Fig. 3

Fig. 3 Diagram of tunable pulsed light induced fluorescence kinetics measurement system.

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3.2 Comparative experiment for photosynthetic rate

Photosynthetic oxygen evolution rate measurement: The oxygen evolution was measured by Chlorolab2 every 120 seconds, and the total oxygen evolution rate was obtained by calculating the slope of oxygen evolution amount of adjacent moments. The net photosynthetic oxygen evolution rate PO2 was obtained by deducting the respiration rate (i.e., oxygen consumption rate under dark adaptation).

Photosynthetic electron transport rate measurement: the fluorescence kinetics curve was measured every 5 seconds, and the instantaneous photosynthetic electron transport rate was calculated from the curve. The instantaneous photosynthetic electron transport rates were averaged every 120 seconds to get a photosynthetic electron transport rate Pe that corresponding to PO2 that measured by Chlorolab2.

Comparative experiments: Chlorella pyrenoidosa was used as the experimental subject. Under different DCMU stress conditions, different culture light and different nutrient (nitrogen) concentrations, the Chlorolab2 and TPLIF system were used to measure the photosynthetic oxygen evolution rate PO2 and photosynthetic electron transport rate Pe respectively. Each sample was measured under eight ambient light intensities, and the results were analyzed, as shown in Fig. 4. In order to ensure the consistency of experimental samples and conditions, the following steps were taken. Firstly, the samples came from the same source. Secondly, the sample pools and ambient light sources used in Chlorolab2 and TPLIF system were respectively of the same specification, and the ambient light sources of the two instruments were synchronized by computer.

 figure: Fig. 4

Fig. 4 Procedure of comparative experiment for phytoplankton photosynthetic rate.

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4. Results and discussion

4.1 DCMU stress experiment

DCMU is an electron transport inhibitor that can inhibit the photosynthesis by inhibiting the QA reoxidation. Sample groups with DCMU concentrations of 30μmol/L, 50μmol/L, 80μmol/L, 100μmol/L were prepared by adding DCMU to the Chlorella pyrenoidosa solution. The Chlorella pyrenoidosa in the solution is in logarithmic phase. A control group without DCMU was also prepared. Each sample group contained three parallel samples. The Chlorolab2 and TPLIF system were used to record the photosynthetic oxygen evolution rate and the electron transport rate respectively under the eight ambient light intensities of 9L, 19L, 29L, 40L, 62L, 74L, 87L,100L for each sample group.

As shown in Fig. 5(a) and Fig. 5(b), as the concentration of DCMU increases, the phytoplankton photosynthesis is increasingly stressed. The photosynthetic rate curves and photosynthetic electron transfer rate curves have the same variation trend and the correlation coefficient R2 between PO2 and Pe is 0.934, as shown in Fig. 5(d). Under an ambient light intensity of 100L, PO2 and Pe decrease obviously when the DCMU concentration is lower than 80μmol/L, while when the DCMU concentration is higher than 80μmol/L, PO2 and Pe almost remain unchanged, indicating that the DCMU stress reaches a threshold, as shown in Fig. 5(c).

 figure: Fig. 5

Fig. 5 Results of phytoplankton photosynthetic rate under DCMU stress (a)Photosynthetic oxygen evolution rate curves (b)Photosynthetic electron transport rate curves (c)Change of PO2 and Pe versus DCMU concentration (d)Correlation between PO2 and Pe.

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4.2 Culture light intensity experiment

Light is the energy source of photosynthesis. Sample groups were cultured under seven light intensities of 60L, 100L, 150L, 200L, 300L, 500L, and 700L respectively. Each sample group contained three parallel samples. After 5-day culture, The Chlorolab2 and TPLIF system were used to respectively record the photosynthetic oxygen evolution rate and the electron transport rate under the eight ambient light intensities of 9L, 19L, 29L, 40L, 62L, 74L, 87L, 100L for each sample group.

It shows in Fig. 6 that the photosynthetic oxygen evolution rate and photosynthetic electron transport rate possess the same variation trend under all the seven culture light intensities. The photosynthetic rate reaches its maximum under 150L culture light intensity, but begins to decrease under higher culture light intensity because of photoinhibition, which is consist with reported results [9]. The correlation coefficient R2 between PO2 and Pe is 0.957.

 figure: Fig. 6

Fig. 6 Results of phytoplankton photosynthetic rate under different culture light intensities (a)Photosynthetic oxygen evolution rate curves (b)Photosynthetic electron transport rate curves (c)Change of PO2 and Pe versus DCMU concentration (d)Correlation between PO2 and Pe.

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4.3 Nutrient (nitrogen) culture experiment

Nitrogen and phosphorus are essential nutrients for the phytoplankton, and nitrogen is crucial for the production of chlorophyll. Six sample groups with nutrient (nitrogen) concentrations of 16mg/L, 32mg/L, 64mg/L, 128mg/L, 228mg/L, 512mg/L were prepared respectively by regulating the nitrogen concentration in BG11 culture medium using NaNO3. Chlorolab2 and TPLIF system were used respectively to record the photosynthetic oxygen evolution rate and the electron transport rate under the eight ambient light intensities of 9L, 19L, 29L, 40L, 62L, 74L, 87L, 100L for each sample group.

As shown in Fig. 7, the photosynthetic oxygen evolution rate and photosynthetic electron transport rate also possess the same variation trend under all nutrient (nitrogen) concentrations, and the correlation coefficient R2 between PO2 and Pe is 0.955. Under nitrogen deficiency (16mg/L) condition, the photosynthetic rate is low, indicating the photosynthesis is limited. The photosynthetic rate increased significantly with the increase of nitrogen concentration (32mg/L, 64mg/L). However, the photosynthetic rate begins to decrease when the nitrogen concentration is higher than 128mg/L. The reason for the decrease of photosynthetic rate under high nitrogen concentration is unknown and needs further study.

 figure: Fig. 7

Fig. 7 Results of phytoplankton photosynthetic rate under different nutrient concentrations (a) Photosynthetic oxygen evolution rate curves (b) Photosynthetic electron transport rate curves (c) Change of PO2 and Pe versus DCMU concentration (d)Correlation between PO2 and Pe.

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5. Conclusion

On the basis of “Bio-Optical” model, a photosynthetic rate measurement method based on tunable pulsed light induced fluorescence kinetics was put forward in this paper. The phytoplankton photosynthetic rate was evaluated by photosynthetic electron transport rate. Comparative experiment results showed that the photosynthetic electron transport rate measured by fluorescence kinetic method under different conditions of DCMU, culture light and nutrients (nitrogen) were consistent with the photosynthetic oxygen evolution rate measured by oxygen evolution method, and the correlation coefficients R2 were above 0.934.

The proposed method possesses the advantages of rapid measurement, non-pretreatment, non-pollution and non-invasion, and breaks through the limitation of gas exchange method, which requires “sampling-incubation-offline analysis”. It is an effective method to measure the phytoplankton photosynthetic rate in situ, thus possesses a broad prospect for applications in water ecological environment monitoring, marine resource assessment and global climate change prediction.

Funding

The National Key Research and Development Program of China (2016YFC1400600); The Open Fund of Qingdao National Laboratory for Marine Science and Technology (QNLM2016ORP0312); National “863” Program of China (2014AA06A509); Natural National Science Foundation of China (31400317); Natural Science Foundation of Anhui Province (1708085QD87).

References and links

1. C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004). [CrossRef]   [PubMed]  

2. P. G. Falkowski and Z. Kolber, “Estimating phytoplankton photosynthesis by active fluorescence,” Carbon Cycle (1992).

3. U. Schreiber, U. Schliwa, and W. Bilger, “Continuous recording of photochemical and non-photochemical chlorophyll fluorescence quenching with a new type of modulation fluorometer,” Photosynth. Res. 10(1-2), 51–62 (1986). [CrossRef]   [PubMed]  

4. Z. S. Kolber and P. G. Falkowski, “Fast Repetition Rate (FRR) Fluorometer For Making In Situ Measurements Of Primary Productivity,” Proc. IEEE 2, 637–641 (1992).

5. P. G. Falkowski and J. A. Raven, “Aquatic photosynthesis: (second edition)”. Freshwater Biology, 53(2), 423–423 (2013).

6. Z. S. Qin, N. J. Zhao, G. F. Yin, C. Y. Shi, T. T. Gan, X. Xiao, J. B. Duan, X. L. Zhang, S. Chen, J. G. Liu, and W. Q. Liu, “Inversion Method of Plant Photosynthesis Parameter Based on Fast Phase and Relaxation Fluorescence Kinetics,” Acta Opt. Sin. 37(7), 0730002 (2017).

7. D. J. Suggett, H. L. MacIntyre, T. M. Kana, and R. J. Geider, “Comparing electron transport with gas exchange: parameterising exchange rates between alternative photosynthetic currencies for eukaryotic phytoplankton,” Aquat. Microb. Ecol. 56(2–3), 147–162 (2009). [CrossRef]  

8. C. Y. Shi, Y. J. Zhang, G. F. Yin, N. J. Zhao, J. B. Duan, X. H. Qiu, L. Fang, X. Xiao, and W. Q. Liu, “Determining the optimal excitation condition of high-frequency flash method for algae photosynthetic parameters measurement,” Guangzi Xuebao 44(2), 5–9 (2015).

9. Z. R. Ouyang, X. B. Wen, Y. H. Geng, H. Mei, H. J. Hu, G. Y. Zhang, and Y. J. Li, “The effects of light intensities, temperatures, pH and salinities on photosynthesis of chlorella,” J. Wuhan Bot. Res. 28(1), 49–55 (2010).

References

  • View by:

  1. C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
    [Crossref] [PubMed]
  2. P. G. Falkowski and Z. Kolber, “Estimating phytoplankton photosynthesis by active fluorescence,” Carbon Cycle (1992).
  3. U. Schreiber, U. Schliwa, and W. Bilger, “Continuous recording of photochemical and non-photochemical chlorophyll fluorescence quenching with a new type of modulation fluorometer,” Photosynth. Res. 10(1-2), 51–62 (1986).
    [Crossref] [PubMed]
  4. Z. S. Kolber and P. G. Falkowski, “Fast Repetition Rate (FRR) Fluorometer For Making In Situ Measurements Of Primary Productivity,” Proc. IEEE 2, 637–641 (1992).
  5. P. G. Falkowski and J. A. Raven, “Aquatic photosynthesis: (second edition)”. Freshwater Biology, 53(2), 423–423 (2013).
  6. Z. S. Qin, N. J. Zhao, G. F. Yin, C. Y. Shi, T. T. Gan, X. Xiao, J. B. Duan, X. L. Zhang, S. Chen, J. G. Liu, and W. Q. Liu, “Inversion Method of Plant Photosynthesis Parameter Based on Fast Phase and Relaxation Fluorescence Kinetics,” Acta Opt. Sin. 37(7), 0730002 (2017).
  7. D. J. Suggett, H. L. MacIntyre, T. M. Kana, and R. J. Geider, “Comparing electron transport with gas exchange: parameterising exchange rates between alternative photosynthetic currencies for eukaryotic phytoplankton,” Aquat. Microb. Ecol. 56(2–3), 147–162 (2009).
    [Crossref]
  8. C. Y. Shi, Y. J. Zhang, G. F. Yin, N. J. Zhao, J. B. Duan, X. H. Qiu, L. Fang, X. Xiao, and W. Q. Liu, “Determining the optimal excitation condition of high-frequency flash method for algae photosynthetic parameters measurement,” Guangzi Xuebao 44(2), 5–9 (2015).
  9. Z. R. Ouyang, X. B. Wen, Y. H. Geng, H. Mei, H. J. Hu, G. Y. Zhang, and Y. J. Li, “The effects of light intensities, temperatures, pH and salinities on photosynthesis of chlorella,” J. Wuhan Bot. Res. 28(1), 49–55 (2010).

2017 (1)

Z. S. Qin, N. J. Zhao, G. F. Yin, C. Y. Shi, T. T. Gan, X. Xiao, J. B. Duan, X. L. Zhang, S. Chen, J. G. Liu, and W. Q. Liu, “Inversion Method of Plant Photosynthesis Parameter Based on Fast Phase and Relaxation Fluorescence Kinetics,” Acta Opt. Sin. 37(7), 0730002 (2017).

2015 (1)

C. Y. Shi, Y. J. Zhang, G. F. Yin, N. J. Zhao, J. B. Duan, X. H. Qiu, L. Fang, X. Xiao, and W. Q. Liu, “Determining the optimal excitation condition of high-frequency flash method for algae photosynthetic parameters measurement,” Guangzi Xuebao 44(2), 5–9 (2015).

2010 (1)

Z. R. Ouyang, X. B. Wen, Y. H. Geng, H. Mei, H. J. Hu, G. Y. Zhang, and Y. J. Li, “The effects of light intensities, temperatures, pH and salinities on photosynthesis of chlorella,” J. Wuhan Bot. Res. 28(1), 49–55 (2010).

2009 (1)

D. J. Suggett, H. L. MacIntyre, T. M. Kana, and R. J. Geider, “Comparing electron transport with gas exchange: parameterising exchange rates between alternative photosynthetic currencies for eukaryotic phytoplankton,” Aquat. Microb. Ecol. 56(2–3), 147–162 (2009).
[Crossref]

2004 (1)

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

1992 (1)

Z. S. Kolber and P. G. Falkowski, “Fast Repetition Rate (FRR) Fluorometer For Making In Situ Measurements Of Primary Productivity,” Proc. IEEE 2, 637–641 (1992).

1986 (1)

U. Schreiber, U. Schliwa, and W. Bilger, “Continuous recording of photochemical and non-photochemical chlorophyll fluorescence quenching with a new type of modulation fluorometer,” Photosynth. Res. 10(1-2), 51–62 (1986).
[Crossref] [PubMed]

Bilger, W.

U. Schreiber, U. Schliwa, and W. Bilger, “Continuous recording of photochemical and non-photochemical chlorophyll fluorescence quenching with a new type of modulation fluorometer,” Photosynth. Res. 10(1-2), 51–62 (1986).
[Crossref] [PubMed]

Bullister, J. L.

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

Chen, S.

Z. S. Qin, N. J. Zhao, G. F. Yin, C. Y. Shi, T. T. Gan, X. Xiao, J. B. Duan, X. L. Zhang, S. Chen, J. G. Liu, and W. Q. Liu, “Inversion Method of Plant Photosynthesis Parameter Based on Fast Phase and Relaxation Fluorescence Kinetics,” Acta Opt. Sin. 37(7), 0730002 (2017).

Duan, J. B.

Z. S. Qin, N. J. Zhao, G. F. Yin, C. Y. Shi, T. T. Gan, X. Xiao, J. B. Duan, X. L. Zhang, S. Chen, J. G. Liu, and W. Q. Liu, “Inversion Method of Plant Photosynthesis Parameter Based on Fast Phase and Relaxation Fluorescence Kinetics,” Acta Opt. Sin. 37(7), 0730002 (2017).

C. Y. Shi, Y. J. Zhang, G. F. Yin, N. J. Zhao, J. B. Duan, X. H. Qiu, L. Fang, X. Xiao, and W. Q. Liu, “Determining the optimal excitation condition of high-frequency flash method for algae photosynthetic parameters measurement,” Guangzi Xuebao 44(2), 5–9 (2015).

Falkowski, P. G.

Z. S. Kolber and P. G. Falkowski, “Fast Repetition Rate (FRR) Fluorometer For Making In Situ Measurements Of Primary Productivity,” Proc. IEEE 2, 637–641 (1992).

Fang, L.

C. Y. Shi, Y. J. Zhang, G. F. Yin, N. J. Zhao, J. B. Duan, X. H. Qiu, L. Fang, X. Xiao, and W. Q. Liu, “Determining the optimal excitation condition of high-frequency flash method for algae photosynthetic parameters measurement,” Guangzi Xuebao 44(2), 5–9 (2015).

Feely, R. A.

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

Gan, T. T.

Z. S. Qin, N. J. Zhao, G. F. Yin, C. Y. Shi, T. T. Gan, X. Xiao, J. B. Duan, X. L. Zhang, S. Chen, J. G. Liu, and W. Q. Liu, “Inversion Method of Plant Photosynthesis Parameter Based on Fast Phase and Relaxation Fluorescence Kinetics,” Acta Opt. Sin. 37(7), 0730002 (2017).

Geider, R. J.

D. J. Suggett, H. L. MacIntyre, T. M. Kana, and R. J. Geider, “Comparing electron transport with gas exchange: parameterising exchange rates between alternative photosynthetic currencies for eukaryotic phytoplankton,” Aquat. Microb. Ecol. 56(2–3), 147–162 (2009).
[Crossref]

Geng, Y. H.

Z. R. Ouyang, X. B. Wen, Y. H. Geng, H. Mei, H. J. Hu, G. Y. Zhang, and Y. J. Li, “The effects of light intensities, temperatures, pH and salinities on photosynthesis of chlorella,” J. Wuhan Bot. Res. 28(1), 49–55 (2010).

Gruber, N.

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

Hu, H. J.

Z. R. Ouyang, X. B. Wen, Y. H. Geng, H. Mei, H. J. Hu, G. Y. Zhang, and Y. J. Li, “The effects of light intensities, temperatures, pH and salinities on photosynthesis of chlorella,” J. Wuhan Bot. Res. 28(1), 49–55 (2010).

Kana, T. M.

D. J. Suggett, H. L. MacIntyre, T. M. Kana, and R. J. Geider, “Comparing electron transport with gas exchange: parameterising exchange rates between alternative photosynthetic currencies for eukaryotic phytoplankton,” Aquat. Microb. Ecol. 56(2–3), 147–162 (2009).
[Crossref]

Key, R. M.

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

Kolber, Z. S.

Z. S. Kolber and P. G. Falkowski, “Fast Repetition Rate (FRR) Fluorometer For Making In Situ Measurements Of Primary Productivity,” Proc. IEEE 2, 637–641 (1992).

Kozyr, A.

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

Lee, K.

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

Li, Y. J.

Z. R. Ouyang, X. B. Wen, Y. H. Geng, H. Mei, H. J. Hu, G. Y. Zhang, and Y. J. Li, “The effects of light intensities, temperatures, pH and salinities on photosynthesis of chlorella,” J. Wuhan Bot. Res. 28(1), 49–55 (2010).

Liu, J. G.

Z. S. Qin, N. J. Zhao, G. F. Yin, C. Y. Shi, T. T. Gan, X. Xiao, J. B. Duan, X. L. Zhang, S. Chen, J. G. Liu, and W. Q. Liu, “Inversion Method of Plant Photosynthesis Parameter Based on Fast Phase and Relaxation Fluorescence Kinetics,” Acta Opt. Sin. 37(7), 0730002 (2017).

Liu, W. Q.

Z. S. Qin, N. J. Zhao, G. F. Yin, C. Y. Shi, T. T. Gan, X. Xiao, J. B. Duan, X. L. Zhang, S. Chen, J. G. Liu, and W. Q. Liu, “Inversion Method of Plant Photosynthesis Parameter Based on Fast Phase and Relaxation Fluorescence Kinetics,” Acta Opt. Sin. 37(7), 0730002 (2017).

C. Y. Shi, Y. J. Zhang, G. F. Yin, N. J. Zhao, J. B. Duan, X. H. Qiu, L. Fang, X. Xiao, and W. Q. Liu, “Determining the optimal excitation condition of high-frequency flash method for algae photosynthetic parameters measurement,” Guangzi Xuebao 44(2), 5–9 (2015).

MacIntyre, H. L.

D. J. Suggett, H. L. MacIntyre, T. M. Kana, and R. J. Geider, “Comparing electron transport with gas exchange: parameterising exchange rates between alternative photosynthetic currencies for eukaryotic phytoplankton,” Aquat. Microb. Ecol. 56(2–3), 147–162 (2009).
[Crossref]

Mei, H.

Z. R. Ouyang, X. B. Wen, Y. H. Geng, H. Mei, H. J. Hu, G. Y. Zhang, and Y. J. Li, “The effects of light intensities, temperatures, pH and salinities on photosynthesis of chlorella,” J. Wuhan Bot. Res. 28(1), 49–55 (2010).

Millero, F. J.

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

Ono, T.

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

Ouyang, Z. R.

Z. R. Ouyang, X. B. Wen, Y. H. Geng, H. Mei, H. J. Hu, G. Y. Zhang, and Y. J. Li, “The effects of light intensities, temperatures, pH and salinities on photosynthesis of chlorella,” J. Wuhan Bot. Res. 28(1), 49–55 (2010).

Peng, T. H.

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

Qin, Z. S.

Z. S. Qin, N. J. Zhao, G. F. Yin, C. Y. Shi, T. T. Gan, X. Xiao, J. B. Duan, X. L. Zhang, S. Chen, J. G. Liu, and W. Q. Liu, “Inversion Method of Plant Photosynthesis Parameter Based on Fast Phase and Relaxation Fluorescence Kinetics,” Acta Opt. Sin. 37(7), 0730002 (2017).

Qiu, X. H.

C. Y. Shi, Y. J. Zhang, G. F. Yin, N. J. Zhao, J. B. Duan, X. H. Qiu, L. Fang, X. Xiao, and W. Q. Liu, “Determining the optimal excitation condition of high-frequency flash method for algae photosynthetic parameters measurement,” Guangzi Xuebao 44(2), 5–9 (2015).

Rios, A. F.

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

Sabine, C. L.

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

Schliwa, U.

U. Schreiber, U. Schliwa, and W. Bilger, “Continuous recording of photochemical and non-photochemical chlorophyll fluorescence quenching with a new type of modulation fluorometer,” Photosynth. Res. 10(1-2), 51–62 (1986).
[Crossref] [PubMed]

Schreiber, U.

U. Schreiber, U. Schliwa, and W. Bilger, “Continuous recording of photochemical and non-photochemical chlorophyll fluorescence quenching with a new type of modulation fluorometer,” Photosynth. Res. 10(1-2), 51–62 (1986).
[Crossref] [PubMed]

Shi, C. Y.

Z. S. Qin, N. J. Zhao, G. F. Yin, C. Y. Shi, T. T. Gan, X. Xiao, J. B. Duan, X. L. Zhang, S. Chen, J. G. Liu, and W. Q. Liu, “Inversion Method of Plant Photosynthesis Parameter Based on Fast Phase and Relaxation Fluorescence Kinetics,” Acta Opt. Sin. 37(7), 0730002 (2017).

C. Y. Shi, Y. J. Zhang, G. F. Yin, N. J. Zhao, J. B. Duan, X. H. Qiu, L. Fang, X. Xiao, and W. Q. Liu, “Determining the optimal excitation condition of high-frequency flash method for algae photosynthetic parameters measurement,” Guangzi Xuebao 44(2), 5–9 (2015).

Suggett, D. J.

D. J. Suggett, H. L. MacIntyre, T. M. Kana, and R. J. Geider, “Comparing electron transport with gas exchange: parameterising exchange rates between alternative photosynthetic currencies for eukaryotic phytoplankton,” Aquat. Microb. Ecol. 56(2–3), 147–162 (2009).
[Crossref]

Tilbrook, B.

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

Wallace, D. W.

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

Wanninkhof, R.

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

Wen, X. B.

Z. R. Ouyang, X. B. Wen, Y. H. Geng, H. Mei, H. J. Hu, G. Y. Zhang, and Y. J. Li, “The effects of light intensities, temperatures, pH and salinities on photosynthesis of chlorella,” J. Wuhan Bot. Res. 28(1), 49–55 (2010).

Wong, C. S.

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

Xiao, X.

Z. S. Qin, N. J. Zhao, G. F. Yin, C. Y. Shi, T. T. Gan, X. Xiao, J. B. Duan, X. L. Zhang, S. Chen, J. G. Liu, and W. Q. Liu, “Inversion Method of Plant Photosynthesis Parameter Based on Fast Phase and Relaxation Fluorescence Kinetics,” Acta Opt. Sin. 37(7), 0730002 (2017).

C. Y. Shi, Y. J. Zhang, G. F. Yin, N. J. Zhao, J. B. Duan, X. H. Qiu, L. Fang, X. Xiao, and W. Q. Liu, “Determining the optimal excitation condition of high-frequency flash method for algae photosynthetic parameters measurement,” Guangzi Xuebao 44(2), 5–9 (2015).

Yin, G. F.

Z. S. Qin, N. J. Zhao, G. F. Yin, C. Y. Shi, T. T. Gan, X. Xiao, J. B. Duan, X. L. Zhang, S. Chen, J. G. Liu, and W. Q. Liu, “Inversion Method of Plant Photosynthesis Parameter Based on Fast Phase and Relaxation Fluorescence Kinetics,” Acta Opt. Sin. 37(7), 0730002 (2017).

C. Y. Shi, Y. J. Zhang, G. F. Yin, N. J. Zhao, J. B. Duan, X. H. Qiu, L. Fang, X. Xiao, and W. Q. Liu, “Determining the optimal excitation condition of high-frequency flash method for algae photosynthetic parameters measurement,” Guangzi Xuebao 44(2), 5–9 (2015).

Zhang, G. Y.

Z. R. Ouyang, X. B. Wen, Y. H. Geng, H. Mei, H. J. Hu, G. Y. Zhang, and Y. J. Li, “The effects of light intensities, temperatures, pH and salinities on photosynthesis of chlorella,” J. Wuhan Bot. Res. 28(1), 49–55 (2010).

Zhang, X. L.

Z. S. Qin, N. J. Zhao, G. F. Yin, C. Y. Shi, T. T. Gan, X. Xiao, J. B. Duan, X. L. Zhang, S. Chen, J. G. Liu, and W. Q. Liu, “Inversion Method of Plant Photosynthesis Parameter Based on Fast Phase and Relaxation Fluorescence Kinetics,” Acta Opt. Sin. 37(7), 0730002 (2017).

Zhang, Y. J.

C. Y. Shi, Y. J. Zhang, G. F. Yin, N. J. Zhao, J. B. Duan, X. H. Qiu, L. Fang, X. Xiao, and W. Q. Liu, “Determining the optimal excitation condition of high-frequency flash method for algae photosynthetic parameters measurement,” Guangzi Xuebao 44(2), 5–9 (2015).

Zhao, N. J.

Z. S. Qin, N. J. Zhao, G. F. Yin, C. Y. Shi, T. T. Gan, X. Xiao, J. B. Duan, X. L. Zhang, S. Chen, J. G. Liu, and W. Q. Liu, “Inversion Method of Plant Photosynthesis Parameter Based on Fast Phase and Relaxation Fluorescence Kinetics,” Acta Opt. Sin. 37(7), 0730002 (2017).

C. Y. Shi, Y. J. Zhang, G. F. Yin, N. J. Zhao, J. B. Duan, X. H. Qiu, L. Fang, X. Xiao, and W. Q. Liu, “Determining the optimal excitation condition of high-frequency flash method for algae photosynthetic parameters measurement,” Guangzi Xuebao 44(2), 5–9 (2015).

Acta Opt. Sin. (1)

Z. S. Qin, N. J. Zhao, G. F. Yin, C. Y. Shi, T. T. Gan, X. Xiao, J. B. Duan, X. L. Zhang, S. Chen, J. G. Liu, and W. Q. Liu, “Inversion Method of Plant Photosynthesis Parameter Based on Fast Phase and Relaxation Fluorescence Kinetics,” Acta Opt. Sin. 37(7), 0730002 (2017).

Aquat. Microb. Ecol. (1)

D. J. Suggett, H. L. MacIntyre, T. M. Kana, and R. J. Geider, “Comparing electron transport with gas exchange: parameterising exchange rates between alternative photosynthetic currencies for eukaryotic phytoplankton,” Aquat. Microb. Ecol. 56(2–3), 147–162 (2009).
[Crossref]

Guangzi Xuebao (1)

C. Y. Shi, Y. J. Zhang, G. F. Yin, N. J. Zhao, J. B. Duan, X. H. Qiu, L. Fang, X. Xiao, and W. Q. Liu, “Determining the optimal excitation condition of high-frequency flash method for algae photosynthetic parameters measurement,” Guangzi Xuebao 44(2), 5–9 (2015).

J. Wuhan Bot. Res. (1)

Z. R. Ouyang, X. B. Wen, Y. H. Geng, H. Mei, H. J. Hu, G. Y. Zhang, and Y. J. Li, “The effects of light intensities, temperatures, pH and salinities on photosynthesis of chlorella,” J. Wuhan Bot. Res. 28(1), 49–55 (2010).

Photosynth. Res. (1)

U. Schreiber, U. Schliwa, and W. Bilger, “Continuous recording of photochemical and non-photochemical chlorophyll fluorescence quenching with a new type of modulation fluorometer,” Photosynth. Res. 10(1-2), 51–62 (1986).
[Crossref] [PubMed]

Proc. IEEE (1)

Z. S. Kolber and P. G. Falkowski, “Fast Repetition Rate (FRR) Fluorometer For Making In Situ Measurements Of Primary Productivity,” Proc. IEEE 2, 637–641 (1992).

Science (1)

C. L. Sabine, R. A. Feely, N. Gruber, R. M. Key, K. Lee, J. L. Bullister, R. Wanninkhof, C. S. Wong, D. W. Wallace, B. Tilbrook, F. J. Millero, T. H. Peng, A. Kozyr, T. Ono, and A. F. Rios, “The oceanic sink for anthropogenic CO2.,” Science 305(5682), 367–371 (2004).
[Crossref] [PubMed]

Other (2)

P. G. Falkowski and Z. Kolber, “Estimating phytoplankton photosynthesis by active fluorescence,” Carbon Cycle (1992).

P. G. Falkowski and J. A. Raven, “Aquatic photosynthesis: (second edition)”. Freshwater Biology, 53(2), 423–423 (2013).

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Figures (7)

Fig. 1
Fig. 1 Diagram of the energy flow in phytoplankton photosynthesis (modified from [5]).
Fig. 2
Fig. 2 Measurement process of tunable pulsed light induced fluorescence kinetics.
Fig. 3
Fig. 3 Diagram of tunable pulsed light induced fluorescence kinetics measurement system.
Fig. 4
Fig. 4 Procedure of comparative experiment for phytoplankton photosynthetic rate.
Fig. 5
Fig. 5 Results of phytoplankton photosynthetic rate under DCMU stress (a)Photosynthetic oxygen evolution rate curves (b)Photosynthetic electron transport rate curves (c)Change of PO2 and Pe versus DCMU concentration (d)Correlation between PO2 and Pe.
Fig. 6
Fig. 6 Results of phytoplankton photosynthetic rate under different culture light intensities (a)Photosynthetic oxygen evolution rate curves (b)Photosynthetic electron transport rate curves (c)Change of PO2 and Pe versus DCMU concentration (d)Correlation between PO2 and Pe.
Fig. 7
Fig. 7 Results of phytoplankton photosynthetic rate under different nutrient concentrations (a) Photosynthetic oxygen evolution rate curves (b) Photosynthetic electron transport rate curves (c) Change of PO2 and Pe versus DCMU concentration (d)Correlation between PO2 and Pe.

Equations (8)

Equations on this page are rendered with MathJax. Learn more.

P e = E × A PSII × T e
F ( t ) = F o + ( F m F o ) ( 1 exp ( σ PSII 0 t E d t ) )
G ( t ) = F o + ( F m F o ) ( exp ( t / τ QA ) )
A PSII = σ PSII / Φ QA
T e = Φ T × Φ P × Φ QA
Φ P = ( F m F o ) / ( F m ' F o ' )
Φ QA = 1 / τ QA
P e = E × A PSII × T e = E × σ PSII × ( F m F o ) / ( F m ' F o ' ) × 1 / τ QA

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