Abstract
Chlorophyll fluorescence constitutes a simple, rapid, and non-invasive means to assess light utilization in Photosystem II (PS II). This study examines aspects relating to the accuracy and applicability of fluorescence for measurement of PS II photochemical quantum yield in intact leaves. A known source of error is fluorescence emission at 730 nm that arises from Photosystem I (PS I). We measured this PS I offset using a dual channel detection system that allows measurement of fluorescence yield in the red (660 nm < F < 710 nm) or far red (F > 710 nm) region of the fluorescence emission spectrum. The magnitude of the PS I offset was equivalent to 30% and 48% of the dark level fluorescence F0 in the far red region for Helianthus annuus and Sorghum bicolor, respectively. The PS I offset was therefore subtracted from fluorescence yields measured in the far red spectral window prior to calculation of PS II quantum yield. Resulting values of PS II quantum yield were consistently higher than corresponding values based on emission in the red region. The basis for this discrepancy lies in the finite optical thickness of the leaf that leads to selective reabsorption by chlorophyll of red fluorescence emission originating in deeper cell layers. Consequently, red fluorescence measurements preferentially sense emission from chloroplasts in the uppermost layer of the leaf where levels of photoprotective nonphotochemical quenching are higher due to increased photon density. It is suggested that far red fluorescence, corrected for the PS I offset, provides the most reliable quantitative basis for calculation of PS II quantum yield because of reduced sensitivity of these measurements to gradients in leaf transmittance and quenching capacity.
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References
Baker NR, Bradbury M, Farage PK, Ireland CR and Long SP (1989) Measurements of the quantum yield of carbon assimilation and chlorophyll fluorescence for assessment of photosynthetic performance of crops in the field. Phil Trans R Soc London B 323: 295–308
Bilger W and Björkman O (1990) Role of the xanthophyll cycle in photoprotection elucidated by measurements of light-induced absorbance changes, fluorescence and photosynthesis in leaves of Hedera canariensis. Photosyth Res 25: 173–185
Bolhàr-Nordenkampf HR and Öquist G (1993) Chlorophyll fluorescence as a tool in photosynthesis research. In: Hall DO, Scurlock JMO, Bolhar-Nordenkampf HR, Leegood RC, Long SP (eds) Photosynthesis and Production in a Changing Environment: A Field and Laboratory Manual, pp 193–205, Chapman & Hall, London
Büchel C and Wilhelm C (1993) In vivo analysis of slow chlorophyll fluorescence induction kinetics in algae: progress, problems and perspectives. Photochem Photobiol 58: 137–148
Butler WL, Kitajima M (1975) Fluorescence quenching in Photosystem II of chloroplasts. Biochim Biophys Acta 376: 116–125
Dau H (1994) Molecular mechanisms and quantitative models of variable Photosystem II fluorescence. Photochem Photobiol 60: 1–23
Demmig B and Björkman O (1987) Comparison of the effect of excessive light on chlorophyll fluorescence (77 K) and photon yield of O2 evolution in leaves of higher plants. Planta 171: 171–184
Edwards GE and Baker NR (1993) Can CO2 assimilation in maize leaves be predicted accurately from chlorophyll fluorescence analysis? Photosynth Res 37: 89–102
Eichelmann H and Laisk A (2000) Cooperation of Photosystems II and I in leaves as analysed by simultaneous measurements of chlorophyll fluorescence and transmittance at 800 nm. Plant Cell Physiol 41: 138–147
Genty B, Briantais JM and Baker NR (1989) The relationship between quantum yield of photosynthetic electron transport and quenching of chlorophyll fluorescence. Biochim Biophys Acta 990: 87–92
Genty B, Wonders J and Baker NR (1990) Non-photochemical quenching of Fo in leaves is emission wavelength dependent: consequences for quenching analysis and its interpretation. Photosynth Res 26: 133–139
Gilmore AM and Yamamoto HY (1991) Zeaxanthin formation and energy-dependent fluorescence quenching in pea chloroplasts under artificially mediated linear and cyclic electron transport. Plant Physiol 96: 635–643
Govindjee (1995) Sixty-three years since Kautsky: Chlorophyll a fluorescence. Aust J Plant Physiol 22: 131–160
Han T, Vogelmann T and Nishio J (1999) Profiles of photosynthetic oxygen-evolution within leaves of Spinacea oleracea. New Phytol 143: 83–92
Holzwarth AR and Roelofs TA (1992) Recent advances in the understanding of chlorophyll excited state dynamics in thylakoid membranes and isolated reaction centre complexes. Photochem Photobiol B Biol 15: 45–62
Hormann H, Neubauer C and Schreiber U (1994) On the relationship between chlorophyll fluorescence quenching and the quantum yield of electron transport in isolated thylakoids. Photosynth Res 40: 93–106
Horton P, Ruban AV and Walters RG (1996) Regulation of light harvesting in green plants. Annu Rev Plant Physiol Plant Mol Biol 47: 655–684
Krause GH and Somersalo S (1989) Fluorescence as a tool in photosynthesis research: Application in studies of photoinhibition, cold acclimation and freezing stress. Phil Trans R Soc London B 323: 281–293
Laisk A and Edwards GE (1997) CO2 and temperature-related induction of photosynthesis in C4 plants: An approach to the hierarchy of rate-limiting processes. Aust J Plant Physiol 24: 505–516
Laisk A and Edwards GE (1998) Oxygen and electron flow in C4 photosynthesis: Mehler reaction, photorespiration and CO2 concentration in bundle sheath. Planta 205: 632–645
Laisk A and Loreto F (1996) Determining photosynthetic parameters from leaf CO2 exchange and chlorophyll fluorescence: Rubisco specificity factor, dark respiration in the light, excitation distribution between photosystems, alternative electron transport and mesophyll diffusion resistance. Plant Physiol 110: 903–912
Laisk A and Oja V (1976) Adaptation of the photosynthetic apparatus to light profile in the leaf. Fiziologija Rastenij (Sov Plant Physiol) 23: 445–451
Laisk A and Oja V (1998) Dynamics of Leaf Photosynthesis Rapid Response Measurements and Their Interpretations, CSIRO, Canberra
Laisk A, Oja V, Rasulov B, Eichelmann H and Sumberg A (1997) Quantum yields and rate constants of photochemical and nonphotochemical excitation quenching. Experiment and model. Plant Physiol 115: 803–815
Lavergne J and Trissl H-W (1995) Theory of fluorescence induction in Photosystem II: Derivation of analytical expressions in a model including exciton-radical-pair equilibrium and restricted energy transfer between photosynthetic units. Biophys J 68: 2474–2492
Loreto F, Di Marco G, Tricoli D and Sharkey TD (1994) Measurements of mesophyll conductance, photosynthetic electron transport and alternative electron sinks of field grown wheat leaves. Photosynth Res 41: 397–403
Öquist G and Chow WS (1992) On the relationship between the quantum yield of Photosystem II electron transport, as determined by chlorophyll fluorescence and the quantum yield of CO2-dependent O2 evolution. Photosynth Res 33: 51–62
Öquist G, Chow WS and Anderson JM (1992) Photoinhibition of photosynthesis represents a mechanism for the long-term regulation of Photosystem II. Planta 186: 450–460
Peterson RB (1991) Effects of O2 and CO2 concentrations on quantum yields of Photosystems I and II in tobacco leaf tissue. Plant Physiol 97: 1388–1394
Peterson RB (1994) Regulation of electron transport in Photosystems I and II in C3, C3-C4, and C4 species of Panicum in response to changing irradiance and O2 levels. Plant Physiol 105: 349–356
Pfündel E (1998) Estimating the contribution of Photosystem I to total leaf chlorophyll fluorescence. Photosynth Res 56: 185–195
Roelofs TA, Lee C-H and Holzwarth AR (1992) Global target analysis of picosecond chlorophyll fluorescence kinetics from pea chloroplasts. Biophys J 61: 1147–1163
Scheibe R (1987) NADP+-malate dehydrogenase in C3-plants: Regulation and role of a light-activated enzyme. Physiol Plant 71: 393–400
Schreiber U, Bilger W and Schliwa U (1986) Continuous recording of photochemical and non-photochemical chlorophyll fluorescence quenching with a new type of modulation fluorometer. Photosynth Res 10: 51–62
Schreiber U, Bilger W and Neubauer C (1994) Chlorophyll fluorescence as a nonintrusive indicator for rapid assessment of in vivo photosynthesis. Ecol Studies 100: 49–70
Seaton GGR and Walker DA (1990) Chlorophyll fluorescence as a measure of photosynthetic carbon assimilation. Proc R Soc London B 242: 29–35
Seaton GGR and Walker DA (1992) Validating chlorophyll fluorescence measures of efficiency: Observations on fluorimetric estimation of photosynthetic rate. Proc R Soc London B 249:41–47
Sharp RE, Matthews MA and Boyer JS (1984) Kok effect and the quantum yield of photosynthesis. Plant Physiol 75: 95–101
Siebke K and Weis E (1995) Imaging of chlorophyll-a-fluorescence in leaves: Topography of photosynthetic oscillations in leaves of Glechoma hederacea. Photosynth Res 45: 225–237
Terashima I, Wong S-C, Osmond CB and Farquhar GD (1988) Characterisation of non-uniform photosynthesis induced by abscisic acid in leaves having different mesophyll anatomies. Plant Cell Physiol 29: 385–394
Vernotte C, Etienne AL and Briantais J-M (1979) Quenching of the system 2 chlorophyll fluorescence by the plastoquinone pool. Biochim Biophys Acta 545: 519–527
Vogelmann TC (1993) Plant tissue optics. Annu Rev Plant Physiol Plant Mol Biol 44: 231–251
Vogelmann TC and Han T (2000) Measurement of gradients of absorbed light in spinach leaves from chlorophyll fluorescence profiles. Plant Cell Environ 23: 1303–1311
Walters RG and Horton P (1991) Resolution of components of non-photochemical chlorophyll fluorescence quenching in barley leaves. Photosynth Res 27: 121–133
Weis E and Berry JA (1987) Quantum efficiency of Photosystem II in relation to ‘energy’ dependent quenching of chlorophyll fluorescence. Biochim Biophys Acta 894: 198–208
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Peterson, R.B., Oja, V. & Laisk, A. Chlorophyll fluorescence at 680 and 730 nm and leaf photosynthesis. Photosynthesis Research 70, 185–196 (2001). https://doi.org/10.1023/A:1017952500015
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DOI: https://doi.org/10.1023/A:1017952500015