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A new optical leaf‐clip meter for simultaneous non‐destructive assessment of leaf chlorophyll and epidermal flavonoids

Zoran G. Cerovic, Guillaume Masdoumier, NaÏma Ben Ghozlen and 1 more

Physiologia Plantarum · 2012

doi:10.1111/j.1399-3054.2012.01639.x

Abstract

We have characterized a new commercial chlorophyll (Chl) and flavonoid (Flav) meter called Dualex 4 Scientific (Dx4). We compared this device to two other Chl meters, the SPAD‐502 and the CCM‐200. In addition, Dx4 was compared to the leaf‐clip Dualex 3 that measures only epidermal Flav. Dx4 is factory‐calibrated to provide a linear response to increasing leaf Chl content in units of µg cm –2 , as opposed to both SPAD‐502 and CCM‐200 that have a non‐linear response to leaf Chl content. Our comparative calibration by Chl extraction confirmed these responses. It seems that the linear response of Dx4 derives from the use of 710 nm as the sampling wavelength for transmittance. The major advantage of Dx4 is its simultaneous assessment of Chl and Flav on the same leaf spot. This allows the generation of the nitrogen balance index (NBI) used for crop surveys and nitrogen nutrition management. The Dx4 leaf clip, that incorporates a GPS receiver, can be useful for non‐destructive estimation of leaf Chl and Flav contents for ecophysiological research and ground truthing of remote sensing of vegetation. In this work, we also propose a consensus equation for the transformation of SPAD units into leaf Chl content, for general use.

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References 44

  1. 1.

    Agati G, Pinelli P, Cortés Ebner S, Romani A, Cartelat A, Cerovic ZG. Non-destructive evaluation of anthocyanins in olive (Olea europaea) fruits by in situ chlorophyll fluorescence spectroscopy. J Agric Food Chem. 2005;53:1354-1363.

    Not in our library10.1021/jf048381d

  2. 2.

    Arnon DI. Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol. 1949;24:1-15.

    Not in our library10.1104/pp.24.1.1

  3. 3.

    Barnes PW, Searles PS, Ballare CL, Ryel RJ, Caldwell MM. Non-invasive measurements of leaf epidermal transmittance of UV radiation using chlorophyll fluorescence: field and laboratory studies. Physiol Plant. 2000;109:274-283.

    Not in our library

  4. 4.

    Bilger W, Veit M, Schreiber L, Schreiber U. Measurement of leaf epidermal transmittance of UV radiation by chlorophyll fluorescence. Physiol Plant. 1997;101:754-763.

    Not in our library

  5. 5.

    Cartelat A, Cerovic ZG, Goulas Y, Meyer S, Lelarge C, Prioul J-L, Barbottin A, Jeuffroy M-H, Gate P, Agati G, Moya I. Optically assessed contents of leaf polyphenolics and chlorophyll as indicators of nitrogen deficiency in wheat (Triticum aestivum L.). Field Crops Res. 2005;91:35-49.

    Not in our library

  6. 6.

    Carter GA, Spiering BA. Optical properties of intact leaves for estimating chlorophyll concentration. J Environ Qual. 2002;31:1424-1432.

    Not in our library10.2134/jeq2002.1424

  7. 7.

    Castelli F, Contillo R, Miceli F. Non-destructive determination of leaf chlorophyll content in four crop species. J Agron Crop Sci. 1996;177:275-283.

    Not in our library

  8. 8.

    Cerovic ZG, Samson G, Morales F, Tremblay N, Moya I. Ultraviolet-induced fluorescence for plant monitoring: present state and prospects. Agronomie: Agric Environ. 1999;19:543-578.

    Not in our library

  9. 9.

    Cerovic ZG, Ounis A, Cartelat A, Latouche G, Goulas Y, Meyer S, Moya I. The use of chlorophyll fluorescence excitation spectra for the nondestructive in situ assessment of UV-absorbing compounds in leaves. Plant Cell Environ. 2002;25:1663-1676.

    Not in our library

  10. 10.

    Coste S, Baraloto C, Leroy C, Marcon E, Renaud A, Richardson AD, Roggy J-C, Schimann H, Uddling J, Hérault B. Assessing foliar chlorophyll contents with the SPAD-502 chlorophyll meter: a calibration test with thirteen tree species of tropical rainforest in French Guiana. Ann Forest Sci. 2010;67–607:1-5.

    Not in our library

  11. 11.

    Esau K. The leaf. In: Plant Anatomy. 1953;411-469.

    Not in our library

  12. 12.

    2011.

    Not in our library

  13. 13.

    Goulas Y, Cerovic ZG, Moya I. Dispositif de Mesure des Caractéristiques D’absorption Lumineuse d’un Echantillon de Tissus Biologique, Procédé de Mesure Associé, et Applications Dans le Domaine de L’analyse Végétale et le Domaine Médical Not Cap. 2001;36.

    Not in our library

  14. 14.

    Goulas Y, Cerovic ZG, Cartelat A, Moya I. Dualex: a new instrument for field measurements of epidermal UV-absorbance by chlorophyll fluorescence. Appl Opt. 2004;43:4488-4496.

    Not in our library10.1364/ao.43.004488

  15. 15.

    Hawkins TS. Modeling the relationship between extractable chlorophyll and SPAD-502 readings for endangered plant species research. J Nat Conserv. 2009;17:123-127.

    Not in our library

  16. 16.

    Hoel BO, Solhaug KA. Effect of irradiance on chlorophyll estimation with the Minolta SPAD-502 leaf chlorophyll meter. Ann Bot. 1998;82:389-392.

    Not in our library

  17. 17.

    Imanishi J, Nakayama A, Suzuki Y, Imanishi A, Ueda N, Morimoto Y, Yoneda M, Eng LE. Nondestructive determination of leaf chlorophyll content in two flowering cherries using reflectance and absorptance spectra. Landscape Ecol Eng. 2010;6:219-234.

    Not in our library

  18. 18.

    Inada K. Studies on a method for determining deepness of green color and chlorophyll content of intact crop leaves and its practical applications. 1. Principle for estimating the deepness of green color and chlorophyll content of whole leaves. Proc Crop Sci Soc Jpn. 1963;32:157-162.

    Not in our library

  19. 19.

    Kolb C, Käser MA, Kopecky J, Zotz G, Riederer M, Pfündel EE. Effects of natural intensities of visible and ultraviolet radiation on epidermal ultraviolet screening and photosynthesis in grape leaves. Plant Physiol. 2001;127:863-875.

    Not in our library

  20. 20.

    Lashbrooke JG, Young PR, Strever AE, Stander C, Vivier MA. The development of a method for the extraction of carotenoids and chlorophylls from grapevine leaves and berries for HPLC profiling. Aust J Grape Wine Res. 2010;16:349-360.

    Not in our library

  21. 21.

    Legendre P, Legendre L. Numerical Ecology. 1998.

    Not in our library

  22. 22.

    Liakopoulos G, Nikolopoulos D, Klouvatou A, Vekkos K-A, Manetas Y, Karabourniotis G. The photoprotective role of epidermal anthocyanins and surface pubescence in young leaves of grapevine (Vitis vinifera). Ann Bot. 2006;98:257-265.

    Not in our library10.1093/aob/mcl097

  23. 23.

    Ling Q, Huang W, Jarvis P. Use of a SPAD-502 meter to measure leaf chlorophyll concentration in Arabidopsis thaliana. Photosynth Res. 2011a;107:209-214.

    Not in our library10.1007/s11120-010-9606-0

  24. 24.

    Ling Q, Huang W, Jarvis P. Erratum: Use of a SPAD-502 meter to measure leaf chlorophyll concentration in Arabidopsis thaliana. Photosynth Res. 2011b;108:89-89.

    Not in our library10.1007/s11120-010-9606-0

  25. 25.

    Marenco RA, Antezana-Vera SA, Nascimento HCS. Relationship between specific leaf area, leaf thickness, leaf water content and SPAD-502 readings in six amazonian tree species. Photosynthetica. 2009;47:184-190.

    Not in our library

  26. 26.

    Markstädter C, Queck I, Baumeister J, Riederer M, Schreiber U, Bilger W. Epidermal transmission of leaves of Vicia faba for UV radiation as determined by two different methods. Photosynth Res. 2001;67:17-25.

    Not in our library10.1023/a:1010676111026

  27. 27.

    Markwell J, Osterman JC, Mitchell JL. Calibration of the Minolta SPAD-502 leaf chlorophyll meter. Photosynth Res. 1995;46:467-472.

    Not in our library10.1007/bf00032301

  28. 28.

    Marquard RD, Tipton JL. Relationship between extractable chlorophyll and an in situ method to estimate leaf greenness. HortScience. 1987;22:1327.

    Not in our library

  29. 29.

    Meyer S, Cerovic ZG, Goulas Y, Montpied P, Desmotes-Mainard S, Bidel L, Moya I, Dreyer E. Relationships between optically assessed polyphenols and chlorophyll content and the dry mass per leaf area ratio of woody plants: a signature of the carbon and nitrogen balance within leaf? Plant Cell Environ. 2006;29:1338-1348.

    Not in our library10.1111/j.1365-3040.2006.01514.x

  30. 30.

    Minolta. Chlorophyll meter SPAD-502. 1989.

    Not in our library

  31. 31.

    Nauš J, Prokopová J, Řebíček J, Špundová M. SPAD chlorophyll meter reading can be pronouncedly affected by chloroplast movement. Photosynth Res. 2010;105:265-271.

    Not in our library10.1007/s11120-010-9587-z

  32. 32.

    Peng S, Garcia FV, Laza RC, Cassman KG. Adjustment of specific leaf weight improves chlorophyll meter's estimation of rice leaf nitrogen concentration. Agron J. 1993;85:987-990.

    Not in our library

  33. 33.

    Pinkard EA, Patel V, Mohammed C. Chlorophyll and nitrogen determination for plantation-grown Eucalyptus nitens and E. globulus using a non-destructive meter. Forest Ecol Man. 2006;223:211-217.

    Not in our library

  34. 34.

    Porra RJ, Thompson WA, Kreidemann PE. Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophyll a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochim Biophys Acta. 1989;975:384-394.

    Not in our library

  35. 35.

    Richardson AD, Duigan SP, Berlyn GP. An evaluation of noninvasive methods to estimate foliar chlorophyll content. New Phytol. 2002;153:185-194.

    Not in our library

  36. 36.

    Tremblay N, Belec C, Jenni S, Fortier E, Mellgren R. The Dualex – a new tool to determine nitrogen sufficiency in broccoli. Acta Hortic. 2009;824:121-131.

    Not in our library

  37. 37.

    Tremblay N, Wang Z, Belec C. Performance of Dualex in spring wheat for crop nitrogen status assessment, yield prediction and estimation of soil nitrate content. J Plant Nutr. 2010;33:57-70.

    Not in our library

  38. 38.

    Tremblay N, Wang Z, Cerovic ZG. Sensing crop nitrogen status with fluorescence indicators. A review. Agron Sust Develop. 2012;32:451-464.

    Not in our library

  39. 39.

    Uddling J, Gelang-Alfredsson J, Piikki K, Pleijel H. Evaluating the relationship between leaf chlorophyll concentration and SPAD-502 chlorophyll meter readings. Photosynth Res. 2007;91:37-46.

    Not in our library10.1007/s11120-006-9077-5

  40. 40.

    Vogelmann TC. Plant tissue optics. Annu Rev Plant Physiol Plant Mol Biol. 1993;44:231-251.

    Not in our library

  41. 41.

    Wellburn AR. The spectral determination of chlorophylls a and b, as well as total carotenoids, using various solvents with spectrophotometers of different resolution. J Plant Physiol. 1994;144:307-313.

    Not in our library

  42. 42.

    Woolley JT. Reflectance and transmittance of light by leaves. Plant Physiol. 1971;47:656-662.

    Not in our library10.1104/pp.47.5.656

  43. 43.

    Yamada N, Fujimura S. Nondestructive measurement of chlorophyll pigment content in plant leaves from three-color reflectance and transmittance. Appl Opt. 1991;30:3964-3973.

    Not in our library10.1364/ao.30.003964

  44. 44.

    Yu H, Wu H-S, Wang Z-J. Evaluation of SPAD and Dualex for in-season corn nitrogen status estimation. Acta Agron Sin. 2010;36:840-847.

    Not in our library

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