Use of in vivo chlorophyll fluorescence to estimate photosynthetic activity and biomass productivity in microalgae grown in different culture systems

In vivo chlorophyll fluorescence associated to Photosystem II is being used to evaluate photosynthetic activity of microalgae grown in different types of photobioreactors; however, controversy on methodology is usual. Several recommendations on the use of chlorophyll fluorescence to estimate electron transport rate and productivity of microalgae grown in thin-layer cascade cultivators and methacrylate cylindrical vessels are included. Different methodologies related to the measure of photosynthetic activity in microalgae are discussed: (1) measurement of light absorption, (2) determination of electron transport rates versus irradiance and (3) use of simplified devices based on pulse amplitude modulated (PAM) fluorescence as Junior PAM or Pocket PAM with optical fiber and optical head as measuring units, respectively. Data comparisons of in vivo chlorophyll fluorescence by using these devices and other PAM fluorometers as Water- PAM in the microalga Chlorella sp. (Chlorophyta) are presented. Estimations of carbon production and productivity by transforming electron transport rate to gross photosynthetic rate (as oxygen evolution) using reported oxygen produced per photons absorbed values and carbon photosynthetic yield based on reported oxygen/carbon ratio are also shown. The limitation of ETR as estimator of photosynthetic and biomass productivity is discussed. Low cost:quality PAMs can promote monitoring of chlorophyll fluorescence in algal biotechnology to estimate the photosynthetic activity and biomass productivity.

[1]  P. Williams,et al.  Overall planktonic oxygen and carbon dioxide metabolisms: the problem of reconciling observations and calculations of photosynthetic quotients , 1991 .

[2]  B. Osborne,et al.  Light and Photosynthesis in Aquatic Ecosystems. , 1985 .

[3]  Mario R. Tredici,et al.  Photobiology of microalgae mass cultures: understanding the tools for the next green revolution , 2010 .

[4]  F. Colijn,et al.  Photosynthetic activity of natural microphytobenthos populations measured by fluorescence (PAM) and 14C-tracer methods: a comparison , 1998 .

[5]  Dale A. Kiefer,et al.  Spectral absorption by marine particles of coastal waters of Baja California1 , 1982 .

[6]  F. Figueroa,et al.  Relations between electron transport rates determined by pulse amplitude modulated chlorophyll fluorescence and oxygen evolution in macroalgae under different light conditions , 2004, Photosynthesis Research.

[7]  C. Wilhelm,et al.  In vivo ANALYSIS OF SLOW CHLOROPHYLL FLUORESCENCE INDUCTION KINETICS IN ALGAE: PROGRESS, PROBLEMS AND PERSPECTIVES , 1993 .

[8]  Annick Bricaud,et al.  Optical efficiency factors of some phytoplankters1 , 1983 .

[9]  M. Curie.  Parameters of photosynthesis: definitions, theory and interpretation of results , 1997 .

[10]  Geir Johnsen,et al.  Biooptical characteristics of PSII and PSI in 33 species (13 pigment groups) of marine phytoplankton, and the relevance for pulse‐amplitude‐modulated and fast‐repetition‐rate fluorometry 1 , 2007 .

[11]  Giuseppe Torzillo,et al.  Use of chlorophyll fluorescence to estimate the effect of photoinhibition in outdoor cultures ofSpirulina platensis , 1994, Journal of Applied Phycology.

[12]  J. Kopecký,et al.  Photoadaptation of two members of the Chlorophyta (Scenedesmus and Chlorella) in laboratory and outdoor cultures: changes in chlorophyll fluorescence quenching and the xanthophyll cycle , 1999, Planta.

[13]  A. Weidemann,et al.  Quantifying absorption by aquatic particles: A multiple scattering correction for glass-fiber filters , 1993 .

[14]  K. Lüning,et al.  Action spectra and spectral quantum yield of photosynthesis in marine macroalgae with thin and thick thalli , 1985 .

[15]  J. Kromkamp,et al.  Light dependence of quantum yields for PSII charge separation and oxygen evolution in eucaryotic algae , 1998 .

[16]  P. Kroth,et al.  In diatoms, the transthylakoid proton gradient regulates the photoprotective non-photochemical fluorescence quenching beyond its control on the xanthophyll cycle. , 2006, Plant & cell physiology.

[17]  R. Ritchie.  Fitting light saturation curves measured using modulated fluorometry , 2008, Photosynthesis Research.

[18]  J. Peeters,et al.  A model for the relationship between light intensity and the rate of photosynthesis in phytoplankton , 1988 .

[19]  H. Maske,et al.  Quantitative in vivo absorption spectra of phytoplankton: Detrital absorption and comparison with fluorescence excitation spectra' , 1987 .

[20]  W. Bilger,et al.  Continuous recording of photochemical and non-photochemical chlorophyll fluorescence quenching with a new type of modulation fluorometer , 2004, Photosynthesis Research.

[21]  Hugh L. MacIntyre,et al.  PHOTOACCLIMATION OF PHOTOSYNTHESIS IRRADIANCE RESPONSE CURVES AND PHOTOSYNTHETIC PIGMENTS IN MICROALGAE AND CYANOBACTERIA 1 , 2002 .

[22]  Govindjee,et al.  Spectral characteristic of fluorescence induction in a model cyanobacterium, Synechococcus sp. (PCC 7942). , 2009, Biochimica et biophysica acta.

[23]  C. Wilhelm,et al.  Bio-optical modelling of oxygen evolution using in vivo fluorescence: Comparison of measured and calculated photosynthesis/irradiance (P-I) curves in four representative phytoplankton species , 2000 .

[24]  Giuseppe Torzillo,et al.  In situ monitoring of chlorophyll fluorescence to assess the synergistic effect of low temperature and high irradiance stresses inSpirulina cultures grown outdoors in photobioreactors , 1996, Journal of Applied Phycology.

[25]  F. Figueroa,et al.  Light field in algal canopies: changes in spectral light ratios and growth of Porphyra leucosticta Thur in Le Jol , 1996 .

[26]  W. Dennison,et al.  An in situ study of photosynthetic oxygen exchange and electron transport rate in the marine macroalga Ulva lactuca (Chlorophyta) , 2002, Photosynthesis Research.

[27]  D. Hanelt.  Photoinhibiton of photosynthesis in marine macrophytes of the South China Sea , 1992 .

[28]  J. M. Robinson.  Does O2 photoreduction occur within chloroplasts in vivo , 1988 .

[29]  P. Horton,et al.  Molecular design of the photosystem II light-harvesting antenna: photosynthesis and photoprotection. , 2004, Journal of experimental botany.

[30]  F. G. Figueiras,et al.  Determination of phytoplankton absorption coefficient in natural seawater samples: evidence of a unique equation to correct the pathlength amplification on glass-fiber filters , 1996 .

[31]  M. Badger,et al.  A COMPARISON OF PHOTOSYNTHETIC ELECTRON TRANSPORT RATES IN MACROALGAE MEASURED BY PULSE AMPLITUDE MODULATED CHLOROPHYLL FLUOROMETRY AND MASS SPECTROMETRY , 2001 .

[32]  N. Baker,et al.  Chlorophyll Fluorescence as a Probe of Photosynthetic Productivity , 2004 .

[33]  Astrid Vieler,et al.  Photophysiology and primary production of phytoplankton in freshwater. , 2004, Physiologia plantarum.

[34]  P. Falkowski,et al.  Light Harvesting and Utilization by Phytoplankton , 1986 .

[35]  J. Briantais,et al.  The relationship between the quantum yield of photosynthetic electron transport and quenching of chlorophyll fluorescence , 1989 .

[36]  F. Figueroa,et al.  Effects of ultraviolet radiation and nutrients on the structure-function of phytoplankton in a high mountain lake , 2012, Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology.

[37]  A. Melis,et al.  Dynamics of photosynthetic membrane composition and function , 1991 .

[38]  K. Asada,et al.  THE WATER-WATER CYCLE IN CHLOROPLASTS: Scavenging of Active Oxygens and Dissipation of Excess Photons. , 1999, Annual review of plant physiology and plant molecular biology.

[39]  M. Björk,et al.  A METHODOLOGICAL COMPARISON OF PHOTOSYNTHETIC OXYGEN EVOLUTION AND ESTIMATED ELECTRON TRANSPORT RATE IN TROPICAL ULVA (CHLOROPHYCEAE) SPECIES UNDER DIFFERENT LIGHT AND INORGANIC CARBON CONDITIONS 1 , 2003 .

[40]  A. Vonshak,et al.  Chlorophyll Fluorescence Applications in Microalgal Mass Cultures , 2010 .

[41]  C. Wilhelm,et al.  From photons to biomass and biofuels: evaluation of different strategies for the improvement of algal biotechnology based on comparative energy balances , 2011, Applied Microbiology and Biotechnology.

[42]  Michael A. Borowitzka,et al.  Chlorophyll a Fluorescence in Aquatic Sciences: Methods and Applications , 2010 .

[43]  J. Kromkamp,et al.  Estimating phytoplankton primary production in Lake IJsselmeer (The Netherlands) using variable fluorescence (PAM-FRRF) and C-uptake techniques , 2008 .

[44]  T. Platt,et al.  Modelling Primary Production , 1980 .

[45]  F. Figueroa,et al.  Relationship between bio-optical characteristics and photoinhibition of phytoplankton , 1997 .

[46]  Motoaki Kishino,et al.  Estimation of the spectral absorption coefficients of phytoplankton in the sea , 1985 .

[47]  F. Figueroa,et al.  Acclimation of Red Sea macroalgae to solar radiation: photosynthesis and thallus absorptance , 2009 .

[48]  Z. Dubinsky,et al.  Changes in photosynthetic properties measured by oxygen evolution and variable chlorophyll fluorescence in a simulated entrainment experiment with the cyanobacterium Planktothrix rubescens , 2001, Aquatic Sciences.

[49]  Hugh L. MacIntyre,et al.  Comparing electron transport with gas exchange: parameterising exchange rates between alternative photosynthetic currencies for eukaryotic phytoplankton , 2009 .

[50]  The absorption spectra of suspensions of living micro-organisms. , 1954 .

[51]  C. Wilhelm,et al.  The impact of cell-specific absorption properties on the correlation of electron transport rates measured by chlorophyll fluorescence and photosynthetic oxygen production in planktonic algae. , 2011, Plant physiology and biochemistry : PPB.

[52]  J. Grobbelaar,et al.  Upper limits of photosynthetic productivity and problems of scaling , 2009, Journal of Applied Phycology.

[53]  A. M. Johnston,et al.  Mechanisms of inorganic-carbon acquisition in marine phytoplankton and their implications for the use of other resources , 1991 .

[54]  J. Kromkamp,et al.  The use of variable fluorescence measurements in aquatic ecosystems: differences between multiple and single turnover measuring protocols and suggested terminology , 2003 .

[55]  Marcel Babin,et al.  Real-time coastal observing systems for marine ecosystem dynamics and harmful algal blooms: Theory, instrumentation and modelling , 2008 .

[56]  K. Asada,et al.  Quenching Analysis of Chlorophyll Fluorescence by the Saturation Pulse Method: Particular Aspects Relating to the Study of Eukaryotic Algae and Cyanobacteria , 1995 .

[57]  J. Beardall,et al.  Short-term variations in photosynthetic parameters of Nannochloropsis cultures grown in two types of outdoor mass cultivation systems , 2009 .

[58]  Subba M. Rao.  Algal cultures, analogues of blooms and applications , 2006 .

[59]  J. Beardall,et al.  Assessing Nutrient Status of Microalgae Using Chlorophyll a Fluorescence , 2010 .

[60]  A. Ley,et al.  Absolute absorption cross-sections for Photosystem II and the minimum quantum requirement for photosynthesis in Chlorella vulgaris , 1982 .

[61]  G. Edwards,et al.  Quantum yields of photosystem II electron transport and carbon dioxide fixation in C4 plants. , 1990 .

[62]  Joseph A. Berry,et al.  Quantum efficiency of Photosystem II in relation to ‘energy’-dependent quenching of chlorophyll fluorescence , 1987 .

[63]  Trevor Platt,et al.  Mathematical formulation of the relationship between photosynthesis and light for phytoplankton , 1976 .

[64]  Hugh L. MacIntyre,et al.  Fast repetition rate and pulse amplitude modulation chlorophyll a fluorescence measurements for assessment of photosynthetic electron transport in marine phytoplankton , 2003 .

[65]  U. Schreiber,et al.  Assessment of photosystem II photochemical quantum yield by chlorophyll fluorescence quenching analysis , 1995 .

[66]  Satoru Taguchi,et al.  Variability in chlorophyll a specific absorption coefficient in marine phytoplankton as a function of cell size and irradiance , 2002 .

[67]  S. Beer,et al.  Photosynthetic rates of Ulva (Chlorophyta) measured by pulse amplitude modulated (PAM) fluorometry , 2000 .

[68]  Thomas Oppenländer,et al.  Glossary of Terms Used in Photochemistry , 2007 .

[69]  Hugh L. MacIntyre,et al.  Evaluation of biophysical and optical determinations of light absorption by photosystem II in phytoplankton , 2004 .

[70]  Carlos M. Duarte,et al.  Light harvesting among photosynthetic organisms , 1994 .

[71]  D. Campbell,et al.  Chlorophyll Fluorescence Analysis of Cyanobacterial Photosynthesis and Acclimation , 1998, Microbiology and Molecular Biology Reviews.

[72]  D. Hanelt,et al.  Field Studies of Photoinhibition Show Non-Correlationsbetween Oxygen and Fluorescence Measurements in the Arctic Red Alga Palmaria palmata , 1995 .

[73]  J. Lavaud,et al.  Cyclic electron transfer in photosystem II in the marine diatom Phaeodactylum tricornutum. , 2006, Biochimica et biophysica acta.