Investigating UV screening in leaves by two different types of portable UV fluorimeters reveals in vivo screening by anthocyanins and carotenoids

Two portable instruments, designed to evaluate epidermal UV screening in leaves, were compared: the Dualex and the UV-A-PAM fluorimeter. Both instruments excite chlorophyll fluorescence at the same UV wavelengths but reference excitation is in the red and the blue spectral range in the former and the latter fluorimeter, respectively. When analyzing green leaves, general agreement of the data is obtained with the two instruments. In the presence of anthocyanins, the UV-A-PAM fluorimeter provided higher estimates for epidermal UV transmittance than the Dualex fluorimeter, which was attributed to absorption of blue excitation light by anthocyanins. By comparing data from the instruments, anthocyanin-dependent transmittance of 50% was determined in abaxial sides of some autumn leaves, and also in abaxial sides of tropical shade plants. Further, with leaves of chlorophyll b-less mutants of H. vulgare, unusually high epidermal UV transmittance was detected but this was attributed to the lack of chlorophyll b absorption and, in addition, to absorption of blue radiation by xanthophylls which are not functionally connected to photosystems.

[1]  Stephan D. Flint,et al.  Internal filters: Prospects for UV‐acclimation in higher plants , 1983 .

[2]  D. Wettstein,et al.  Chlorina mutants of barley (Hordeum vulgare L.) , 1985 .

[3]  G. Grammatikopoulos,et al.  Site-dependent differences in transmittance and UV-B-absorbing capacity of isolated leaf epidermes and mesophyll in Urginea maritima (L.) Baker , 1999 .

[4]  C. Ballaré,et al.  Functional significance and induction by solar radiation of ultraviolet-absorbing sunscreens in field-grown soybean crops. , 2000, Plant physiology.

[5]  A. Gallé,et al.  Capacity of protection against ultraviolet radiation in sun and shade leaves of tropical forest plants. , 2003, Functional plant biology : FPB.

[6]  B. Jordan.  The Effects of Ultraviolet-B Radiation on Plants: A Molecular Perspective , 1996 .

[7]  Ismael Moya,et al.  Ultraviolet-induced fluorescence for plant monitoring: present state and prospects , 1999 .

[8]  E. Pfündel,et al.  Effects of natural intensities of visible and ultraviolet radiation on epidermal ultraviolet screening and photosynthesis in grape leaves. , 2001, Plant physiology.

[9]  Janet F. Bornman,et al.  A flavonoid mutant of barley (Hordeum vulgare L.) exhibits increased sensitivity to UV‐B radiation in the primary leaf , 1996 .

[10]  Dugald C. Close,et al.  The ecophysiology of foliar anthocyanin , 2003, The Botanical Review.

[11]  R. Smillie,et al.  Photoabatement by Anthocyanin Shields Photosynthetic Systems from Light Stress , 1999, Photosynthetica.

[12]  H. Behnke,et al.  Caryophyllales : evolution and systematics , 1994 .

[13]  N. Simmonds,et al.  A survey of the anthocyanins of some tropical plants , 1954, Proceedings of the Royal Society of London. Series B - Biological Sciences.

[14]  Geza Hrazdina,et al.  Anthocyanin composition of Brassica oleracea cv. Red Danish , 1977 .

[15]  J. Bornman,et al.  Effects of Ultraviolet-B Radiation on Terrestrial Plants , 1993 .

[16]  Giovanni Agati,et al.  Nondestructive evaluation of anthocyanins in olive (Olea europaea) fruits by in situ chlorophyll fluorescence spectroscopy. , 2005, Journal of agricultural and food chemistry.

[17]  J. Sheahan.  SINAPATE ESTERS PROVIDE GREATER UV-B ATTENUATION THAN FLAVONOIDS IN ARABIDOPSIS THALIANA (BRASSICACEAE) , 1996 .

[18]  E. Govindje,et al.  Sixty-Three Years Since Kautsky: Chlorophyll a Fluorescence , 1995 .

[19]  E. C. Bate-smith.  The phenolic constituents of plants and their taxonomic significance. I. Dicotyledons , 1962 .

[20]  M. Tevini,et al.  THE PROTECTIVE FUNCTION OF THE EPIDERMAL LAYER OF RYE SEEDLINGS AGAINST ULTRAVIOLET‐B RADIATION , 1991 .

[21]  R. J. Porra,et al.  Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy , 1989 .

[22]  S. Reuber,et al.  Tissue localization of phenolic compounds in plants by confocal laser scanning microscopy , 1998 .

[23]  M. Bloom.  EVOLUTION IN THE GENUS RUELLIA (ACANTHACEAE): A DISCUSSION BASED ON FLORAL FLAVONOIDS , 1976 .

[24]  Ismael Moya,et al.  Dual-excitation FLIDAR for the estimation of epidermal UV absorption in leaves and canopies , 2001 .

[25]  T. Mabry,et al.  Chemical Review and Evolutionary Significance of the Betalains , 1994 .

[26]  Govindjee,et al.  Chlorophyll a Fluorescence , 2004, Advances in Photosynthesis and Respiration.

[27]  Jeffrey B. Harborne,et al.  The flavonoids: recent advances. , 1988 .

[28]  W. Bilger,et al.  UV-excited chlorophyll fluorescence as a tool for the assessment of UV-protection by the epidermis of plants. , 2001, Journal of experimental botany.

[29]  Markus Riederer,et al.  UV screening by phenolics in berries of grapevine (Vitis vinifera). , 2003, Functional plant biology : FPB.

[30]  Z. Cerovic,et al.  Simultaneous measurement of changes in red and blue fluorescence in illuminated isolated chloroplasts and leaf pieces: The contribution of NADPH to the blue fluorescence signal , 1993, Photosynthesis Research.

[31]  A. Ruban,et al.  Xanthophylls of the major photosynthetic light‐harvesting complex of plants: identification, conformation and dynamics , 2000, FEBS letters.

[32]  Ulrich Schreiber,et al.  Measurement of leaf epidermal transmittance of UV radiation by chlorophyll fluorescence , 1997 .

[33]  B. McCown,et al.  Physiological significance of anthocyanins during autumnal leaf senescence. , 2001, Tree physiology.

[34]  Grethe Iren A. Borge,et al.  Chlorophyll fluorescence as a tool for non-destructive estimation of anthocyanins and total flavonoids in apples , 2006 .

[35]  U. Schreiber,et al.  UV-A screening in plants determined using a new portable fluorimeter , 2005, Photosynthetica.

[36]  D. Siefermann-Harms.  Carotenoids in photosynthesis. I: Location in photosynthetic membranes and light-harvesting function , 1985 .

[37]  E. C. Bate-smith.  The phenolic constituents of plants and their taxonomic significance , 1968 .

[38]  A. Gazula,et al.  Variety, Shading, and Growth Stage Effects on Pigment Concentrations in Lettuce Grown under Contrasting Temperature Regimens , 2003 .

[39]  Craig S. T. Daughtry,et al.  A new technique to measure the spectral properties of conifer needles , 1989 .

[40]  W. Bilger,et al.  Epidermal transmittance of leaves of Vicia faba for UV radiation as determined by two different methods , 2004, Photosynthesis Research.

[41]  S. Wand,et al.  Anthocyanins in vegetative tissues: a proposed unified function in photoprotection. , 2002, The New phytologist.

[42]  J. R. Price,et al.  A survey of anthocyanins. V. , 1938, The Biochemical journal.

[43]  Florian C. Stintzing,et al.  Functional properties of anthocyanins and betalains in plants, food, and in human nutrition , 2004 .

[44]  Yves Goulas,et al.  Dualex: a new instrument for field measurements of epidermal ultraviolet absorbance by chlorophyll fluorescence. , 2004, Applied optics.

[45]  Timothy M. Collins,et al.  Phylogenetic and Ontogenetic Influences on the Distribution of Anthocyanins and Betacyanins in Leaves of Tropical Plants , 2001, International Journal of Plant Sciences.

[46]  Z. Cerovic,et al.  Optical Properties of Plant Surfaces , 2007 .

[47]  Y. Manetas.  Why some leaves are anthocyanic and why most anthocyanic leaves are red , 2006 .

[48]  M. Seufferheld.  Non-photochemical quenching of chlorophyll a fluorescence: early history and characterization of two xanthophyll-cycle mutants of Chlamydomonas reinhardtii. , 2002, Functional plant biology : FPB.

[49]  Ronald J. Ryel,et al.  Non‐invasive measurements of leaf epidermal transmittance of UV radiation using chlorophyll fluorescence: field and laboratory studies , 2000 .

[50]  H. A. Stafford.  Anthocyanins and betalains: evolution of the mutually exclusive pathways , 1994 .

[51]  R. Bassi,et al.  Chlorophyll-proteins from maize seedlings grown under intermittent light conditions , 1993, Planta.

[52]  K. Gould,et al.  Nature's Swiss Army Knife: The Diverse Protective Roles of Anthocyanins in Leaves , 2004, Journal of biomedicine & biotechnology.

[53]  R. Robinson,et al.  A survey of anthocyanins. II. , 1938, Biochemical Journal.

[54]  H. Lichtenthaler,et al.  Characterization of the laser-induced blue, green and red fluorescence signatures of leaves of wheat and soybean grown under different irradiance. , 1993, Physiologia plantarum.

[55]  R. Cogdell,et al.  Carotenoids in Photosynthesis , 1996, Photochemistry and photobiology.

[56]  S. Jansson,et al.  Screening of chlorina mutants of barley (Hordeum vulgare L.) with antibodies against light-harvesting proteins of PS I and PS II: Absence of specific antenna proteins , 1997, Photosynthesis Research.

[57]  B. Logan,et al.  Chlorophyll a Fluorescence: A Signature of Photosynthesis , 2005 .

[58]  E. Baake,et al.  A quantitative description of fluorescence excitation spectra in intact bean leaves greened under intermittent light , 1990, Photosynthesis Research.

[59]  David W. Lee,et al.  Why leaves turn red in autumn. The role of anthocyanins in senescing leaves of red-osier dogwood. , 2001, Plant physiology.

[60]  J. Moan,et al.  Environmental UV Photobiology , 1993, Springer US.

[61]  E. Pfündel,et al.  Origins of non-linear and dissimilar relationships between epidermal UV absorbance and UV absorbance of extracted phenolics in leaves of grapevine and barley , 2005 .

[62]  M. Pedreño,et al.  Studying the oxidation and the antiradical activity of betalain from beetroot , 2000 .