Abstract
By using a fiber-optic microprobe in combination with a modified PAM Fluorometer, chlorophyll fluorescence yield was measured within leaves with spatial resolution of approximately 20 μm. The new system employs a miniature photomultiplier for detection of the pulse-modulated fluorescence signal received by the 20 μm fiber tip. The obtained signal/noise ratio qualifies for recordings of fluorescence induction kinetics (Kautsky effect), fluorescence quenching by the saturation pulse method and determination of quantum yield of energy conversion at Photosystem II at different sites within a leaf. Examples of the system performance and of practical applications are given. It is demonstrated that the fluorescence rise kinetics are distinctly faster when chloroplasts within the spongy mesophyll are illuminated as compared to palisade chloroplasts. Photoinhibition is shown to affect primarily the quantum yield of the palisade chloroplasts when excessive illumination is applied from the adaxial leaf side. The new system is envisaged to be used in combination with light measurements within leaves for an assessment of the specific contributions of different leaf regions to overall photosynthetic activity and for an integrative modelling of leaf photosynthesis.
Similar content being viewed by others
References
Björkman O and Demmig B (1987) Photon yield of O2 evolution and chlorophyll fluorescence characteristics at 77 K among vascular plants of diverse origins. Planta 170: 489–504
Bornman JF, Vogelmann TC and Martin G (1991) Measurement of chlorophyll fluorescence within leaves with a fiber optic microprobe. Plant Cell Environ 14: 719–725
Chylla RA and Whitmarsch J (1989) Inactive Photosystem II complexes in leaves. Plant Physiol 90: 765–772
Cue M, Vogelmann TC and Smith WK (1991) Chlorophyll and light gradients in sun and shade leaves of Spinacia oleracea. Plant Cell Environ 14: 493–500
Fukshansky L and Martinez v. Remisowsky A (1992) A theoretical study of the light microenvironment in a leaf in relation to photosynthesis. Plant Sci 86: 167–182
Fukshansky L, Martinez v. Remisowsky A, McClendon J, Ritterbusch A, Richter T and Mohr H (1992) Absorption spectra of leaves corrected for scattering and distributional error: A radiative transfer and absorption statistics treatment. Photochem Photobiol 55: 857–869
Genty B, Briantais JM and Baker NR (1989) The relationship between the quantum yield of photosynthetic electron transport and quenching of chlorophyll fluorescence. Biochim Biophys Acta 990: 87–90
Genty B and Meyer S (1995) Quantitative mapping of leaf photosynthesis using chlorophyll fluorescence imaging. Aust J Plant Physiol 22: 277–284
Jørgensen BB and DesMarais DJ (1988) Optical properties of benthic photosynthetic communities: fiber optic studies of cyanobacterial mats. Limnol Oceanogr 33: 99–113
Jørgensen BB, Revsbech NP and Cohen Y (1983) Photosynthesis and structure of benthic microbial mats: Microelectrode and SEM studies of four cyanobacterial communities. Limnol Oceanogr 28: 1075–1093
Kautsky H and Hirsch A (1931) Neue Versuche zur Kohlenstoffassimilation. Naturwissenschaften 19: 964
Kitajima M and Butler WL (1975) Quenching of chlorophyll fluorescence and primary photochemistry in chloroplasts by dibromothymoquinone. Biochim Biophys Acta 376: 105–115
Klimant I, Meyer V and Kühl M (1995) Fiber-optic oxygen microsensors, a new tool in aquatic biology. Limnol Oceanogr 40 (in press)
Krause GH and Weis E (1991) Chlorophyll fluorescence and photosynthesis: The basics. Annu Rev Plant Physiol Plant Mol Biol 42: 313–345
Kühl M and Jørgensen BB (1992) Spectral light measurements in microbenthic phototrophic communities with a fiber-optic microbrobe coupled to a sensitive diode array detector. Limnol Oceanogr 37: 1813–1823
Kühl M, Lassen C and Jørgensen BB (1994) Optical properties of microbial mats: Light measurements with fiber-optic microprobes. In: Stal LJ and Caumette P (eds) Microbial Mats, pp 149–166. Springer Verlag, Berlin, Heidelberg
Martinez v. Remisowsky AJ, McClendon J and Fukshansky L (1992) A comparative study of internal light environment in bifacial leaves of different plants. Bot Acta 105: 362–366
Myers AD, Vogelmann TC and Bornman JF (1994) Epidermal focussing and effects on light utilization in Oxalis acetosella. Physiol Plant 91: 651–656
Powles SB and Björkman O (1982) Photoinhibition of photosynthesis: Effect on chlorophyll fluorescence at 77 K in intact leaves and in chloroplast membranes of Nerium oleander. Planta 156: 97–107
Schreiber U (1986) Detection of rapid induction kinetics with a new type of high-frequency modulated chlorophyll fluorometer. Photosynth Res 9: 261–272
Schreiber U (1994) New emitter-detector-cuvette assembly for measuring modulated chlorophyll fluorescence of highly diluted suspensions in conjunction with the standard PAM fluorometer. Z Naturforsch 49c: 646–656
Schreiber U and Bilger (1993) Progress in chlorophyll fluorescence research: Major developments during the last years in retrospect. Prog Bot 54: 151–173
Schreiber U, Bilger W and Neubauer C (1994) Chlorophyll fluorescence as a non-intrusive indicator for rapid assessment of in vivo photosynthesis. Ecological Studies 100: 49–70
Schreiber U, Fink R and Vidaver W (1977) Chlorophyll fluorescence in whole leaves: Photosynthetic adaptation to contrasting light regimes. Planta 133: 121–129
Schreiber U, Schliwa U and Bilger W (1986) Continuous recording of photochemical and non-photochemical chlorophyll fluorescence quenching with a new type of modulation fluorometer. Photosynth Res 10: 51–61
Siebke K and Weis E (1995) ‘Assimilation images’ of leaves of Glechoma hederacea: Analysis of non-synchronous stomata related oscillations. Planta 196: 155–165
Terashima I and Inoue Y (1985a) Vertical gradient in photosynthetic properties of spinach chloroplasts dependent on intra-leaf light environment. Plant Cell Physiol 26: 781–785
Terashima I and Inoue Y (1985b) Palisade tissue chloroplasts and spongy tissue chloroplasts in spinach: biochemical and ultrastructural differences. Plant Cell Physiol 26: 63–75
Vogelmann TC (1993) Plant tissue optics. Ann Rev Plant Physiol Plant Mol Biol 44: 231–251
Vogelmann TC and Björn LO (1984) Measurement of light gradients and spectral regime in plant tissue with a fibre optic probe. Physiol Plant 60: 361–368
Vogelmann TC, Bornman JF and Josserand S (1989) Photosynthetic light gradients and spectral regime within leaves of Medicago sativa. Phil Trans R Soc London B 323: 411–421
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 J (1987) Quantum efficiency of Photosystem II in relation to energy-dependent quenching of chlorophyll fluorescence. Biochim Biophys Acta 894: 198–208
Author information
Authors and Affiliations
Additional information
This paper is dedicated to Ulrich Heber on the occasion of his 65th birthday, with great respect for his outstanding achievements in photosynthesis research.
Rights and permissions
About this article
Cite this article
Schreiber, U., Kühl, M., Klimant, I. et al. Measurement of chlorophyll fluorescence within leaves using a modified PAM Fluorometer with a fiber-optic microprobe. Photosynth Res 47, 103–109 (1996). https://doi.org/10.1007/BF00017758
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00017758