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      Frequently asked questions about in vivo chlorophyll fluorescence: practical issues

      review-article
      , , , , , , , , , , ,   , , , , , , , , , , , , , , , , ,
      Photosynthesis Research
      Springer Netherlands
      Chlorophyll a fluorescence, Fluorescence imaging, Complementary techniques, Frequently asked questions, Plant stress monitoring, Photosynthesis

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          Abstract

          The aim of this educational review is to provide practical information on the hardware, methodology, and the hands on application of chlorophyll (Chl) a fluorescence technology. We present the paper in a question and answer format like frequently asked questions. Although nearly all information on the application of Chl a fluorescence can be found in the literature, it is not always easily accessible. This paper is primarily aimed at scientists who have some experience with the application of Chl a fluorescence but are still in the process of discovering what it all means and how it can be used. Topics discussed are (among other things) the kind of information that can be obtained using different fluorescence techniques, the interpretation of Chl a fluorescence signals, specific applications of these techniques, and practical advice on different subjects, such as on the length of dark adaptation before measurement of the Chl a fluorescence transient. The paper also provides the physiological background for some of the applied procedures. It also serves as a source of reference for experienced scientists.

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          Chlorophyll fluorescence analysis: a guide to good practice and understanding some new applications.

          Chlorophyll fluorescence is a non-invasive measurement of photosystem II (PSII) activity and is a commonly used technique in plant physiology. The sensitivity of PSII activity to abiotic and biotic factors has made this a key technique not only for understanding the photosynthetic mechanisms but also as a broader indicator of how plants respond to environmental change. This, along with low cost and ease of collecting data, has resulted in the appearance of a large array of instrument types for measurement and calculated parameters which can be bewildering for the new user. Moreover, its accessibility can lead to misuse and misinterpretation when the underlying photosynthetic processes are not fully appreciated. This review is timely because it sits at a point of renewed interest in chlorophyll fluorescence where fast measurements of photosynthetic performance are now required for crop improvement purposes. Here we help the researcher make choices in terms of protocols using the equipment and expertise available, especially for field measurements. We start with a basic overview of the principles of fluorescence analysis and provide advice on best practice for taking pulse amplitude-modulated measurements. We also discuss a number of emerging techniques for contemporary crop and ecology research, where we see continual development and application of analytical techniques to meet the new challenges that have arisen in recent years. We end the review by briefly discussing the emerging area of monitoring fluorescence, chlorophyll fluorescence imaging, field phenotyping, and remote sensing of crops for yield and biomass enhancement.
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            Continuous recording of photochemical and non-photochemical chlorophyll fluorescence quenching with a new type of modulation fluorometer.

            A newly developed fluorescence measuring system is employed for the recording of chlorophyll fluorescence induction kinetics (Kautsky-effect) and for the continuous determination of the photochemical and non-photochemical components of fluorescence quenching. The measuring system, which is based on a pulse modulation principle, selectively monitors the fluorescence yield of a weak measuring beam and is not affected even by extremely high intensities of actinic light. By repetitive application of short light pulses of saturating intensity, the fluorescence yield at complete suppression of photochemical quenching is repetitively recorded, allowing the determination of continuous plots of photochemical quenching and non-photochemical quenching. Such plots are compared with the time courses of variable fluorescence at different intensities of actinic illumination. The differences between the observed kinetics are discussed. It is shown that the modulation fluorometer, in combination with the application of saturating light pulses, provides essential information beyond that obtained with conventional chlorophyll fluorometers.
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              Chlorophyll fluorescence--a practical guide.

              Chlorophyll fluorescence analysis has become one of the most powerful and widely used techniques available to plant physiologists and ecophysiologists. This review aims to provide an introduction for the novice into the methodology and applications of chlorophyll fluorescence. After a brief introduction into the theoretical background of the technique, the methodology and some of the technical pitfalls that can be encountered are explained. A selection of examples is then used to illustrate the types of information that fluorescence can provide.
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                Author and article information

                Contributors
                hazem@kalaji.pl
                gert.schansker@gmail.com
                richard.ladle@ouce.ox.ac.uk
                goltsev@gmail.com , goltsev@biofac.uni-sofia.bg
                karolinabosa@gmail.com
                suleyman.allakhverdiev@gmail.com
                marian.brestic@uniag.sk
                filippo.bussotti@unifi.it
                calatayud_ang@gva.es
                piotr.andrzej.dabrowski@gmail.com
                nshery@yahoo.com
                frrlnz@unife.it
                lucia.guidi@unipi.it
                info@plantlighting.nl
                anjanajajoo@hotmail.com
                misraan@yahoo.co.uk , misra.amarendra@gmail.com
                sergonne@bvg.upv.es
                pcs@unife.it
                penella_con@gva.es
                poli@roanoke.edu
                martina.pollastrini@unifi.it
                romano@biol.uni.lodz.pl
                beata_rutkowska@sggw.pl
                jserodio@ua.pt
                sureshkancherla@rediffmail.com
                wieslaw_szulc@sggw.pl
                tambussi35@yahoo.es
                marcosyanniccari@conicet.gov.ar
                marek.zivcak@uniag.sk
                Journal
                Photosynth Res
                Photosyn. Res
                Photosynthesis Research
                Springer Netherlands (Dordrecht )
                0166-8595
                1573-5079
                15 August 2014
                15 August 2014
                2014
                : 122
                : 2
                : 121-158
                Affiliations
                [ ]Department of Plant Physiology, Faculty of Agriculture and Biology, Warsaw University of Life Sciences – SGGW, Nowoursynowska 159, 02-776 Warsaw, Poland
                [ ]Avenue des Amazones 2, 1226 Chêne-Bougeries, Switzerland
                [ ]Institute of Biological and Health Sciences, Federal University of Alagoas, Praça Afrânio Jorge, s/n, Prado, Maceió, AL Brazil
                [ ]Department of Biophysics and Radiobiology, Faculty of Biology, St. Kliment Ohridski University of Sofia, 8 Dr. Tzankov Blvd., 1164 Sofia, Bulgaria
                [ ]Department of Pomology, Faculty of Horticulture, Biotechnology and Landscape Architecture, Warsaw University of Life Sciences – SGGW, Nowoursynowska 159, 02-776 Warsaw, Poland
                [ ]Institute of Plant Physiology, Russian Academy of Sciences, Botanicheskaya Street 35, Moscow, 127276 Russia
                [ ]Institute of Basic Biological Problems, Russian Academy of Sciences, Pushchino, Moscow Region, 142290 Russia
                [ ]Department of Plant Physiology, Slovak Agricultural University, Tr. A. Hlinku 2, 949 76 Nitra, Slovak Republic
                [ ]Department of Agri-Food Production and Environmental Science (DISPAA), University of Florence, Piazzale delle Cascine 28, 50144 Florence, Italy
                [ ]Departamento de Horticultura, Instituto Valenciano de Investigaciones Agrarias, Ctra. Moncada-Náquera Km 4.5, Moncada, 46113 Valencia, Spain
                [ ]Department of Environmental Improvement, Faculty of Civil and Environmental Engineering, Warsaw University of Life Sciences – SGGW, Nowoursynowska 159, 02-776 Warsaw, Poland
                [ ]Agricultural Botany Department, Faculty of Agriculture, Tanta University, Tanta, Egypt
                [ ]Department of Life Sciences and Biotechnologies, University of Ferrara, Corso Ercole I d’Este 32, 44121 Ferrara, Italy
                [ ]Department of Agriculture, Food and Environment, Via del Borghetto, 80, 56124 Pisa, Italy
                [ ]Plant Lighting BV, Veilingweg 46, 3981 PC Bunnik, The Netherlands
                [ ]School of Life Sciences, Devi Ahilya University, Indore, 452 001 M.P India
                [ ]Centre for Life Sciences, Central University of Jharkhand, Ratu-Lohardaga Road, Ranchi, 835205 India
                [ ]Departamento de Producción vegetal, Universitat Politècnica de València, C de Vera sn, 46022 Valencia, Spain
                [ ]Department of Biology, Roanoke College, 221 College Lane, Salem, VA 24153 USA
                [ ]Department of Ecophysiology and Plant Development, University of Lodz, Banacha 12/16, Lodz, 90-237 Poland
                [ ]Agricultural Chemistry Department, Faculty of Agriculture and Biology, Warsaw University of Life Sciences – SGGW, Nowoursynowska 159, 02-776 Warsaw, Poland
                [ ]Departamento de Biologia, CESAM – Centro de Estudos do Ambiente e do Mar, Universidade de Aveiro, Campus de Santiago, 3810-193 Aveiro, Portugal
                [ ]Directorate of Oil Palm Research, West Godavari Dt., Pedavegi, 534 450 Andhra Pradesh India
                [ ]Institute of Plant Physiology, INFIVE (Universidad Nacional de La Plata – Consejo Nacional de Investigaciones Científicas y Técnicas), Diagonal 113 N°495, 327 La Plata, Argentina
                Article
                24
                10.1007/s11120-014-0024-6
                4210649
                25119687
                292e9490-f688-408f-9ec4-68de9ee50b28
                © The Author(s) 2014

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.

                History
                : 9 February 2014
                : 2 June 2014
                Categories
                Review
                Custom metadata
                © Springer Science+Business Media Dordrecht 2014

                Plant science & Botany
                chlorophyll a fluorescence,fluorescence imaging,complementary techniques,frequently asked questions,plant stress monitoring,photosynthesis

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