Skip to main content
SpringerLink
Log in
Book cover

UV Radiation in Global Climate Change pp 437–457Cite as

Assessment of DNA Damage as a Tool to Measure UV-B Tolerance in Soybean Lines Differing in Foliar Flavonoid Composition

  • Chapter

  • 1419 Accesses

Abstract

Damage to DNA, in the form of cyclobutane pyrimidine dimers (CPD), may occur in soybean (Glycine max (L.) Merr) plants when they are exposed to increasing levels of ultraviolet-B (UV-B) radiation. Flavonoids and other phenolics accumulate in the epidermal layer of leaves and may provide protection for sensitive tissues including DNA molecules. We evaluated the steady state levels of accumulated damage and the protection afforded by flavonoids in two soybean isolines: Clark producing high levels of flavonoids, and Clark-magenta producing extremely low flavonoid levels. Both cultivars were grown in the field under ambient and supplemental UV-B radiation. Leaf tissue was harvested in a diurnal sequence, and the samples were analyzed. Two methods of analysis were used in order to develop a common reference point between the two. In one method, DNA was isolated and treated with UV endonuclease, and the DNA fragments were separated using unidirectional pulsed field electrophoresis and quantified through electronic imaging. In the alternate method, a western blotting procedure, immobilized DNA was reacted with monoclonal antibodies specific to CPD DNA damage. Results were similar in both techniques and show lesion frequency to be low in both isolines. However significant differences were found between cultivars, UV treatments, time of day collected, and levels of PAR. The average level of dimers per megabase for the isoline Clark was ∼4 (with or without supplemental UV), and for Clark-magenta, ∼4 for samples with no supplemental UV and ∼6 for those exposed to supplemental UV radiation. Diurnally, dimer levels were frequently higher in the Clark-magenta isoline, especially when exposed to supplemental UV-B. Both isolines appear to be either well-protected from DNA damage, or repair is efficient enough to minimize biologically significant accumulation of DNA damage. This suggests that protection mechanisms, other than flavonoids alone, contribute to maintenance of DNA integrity in soybean.

This is a preview of subscription content, log in via an institution.

Chapter
USD   29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   219.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • Barnes PW, Flint SD and Caldwell MM (1987) Photosynthesis damage and protective pigments in plants from a latitudinal arctic/alpine gradient exposed to supplemental UV-B radiation in the field. Arctic and Alpine Research 19: 21–27

    Article  Google Scholar 

  • Beerhues L, Robenek H and Wiermann R (1988) Chalcone synthesis from spinach (Spinacia oleracea L.) II. Immunofluorescence and immunogold localization. Planta 173: 544–553

    Article  CAS  Google Scholar 

  • Beggs CJ, Schneider-Ziebert U and Wellmann E (1986) UV-B radiation and adaptive mechanisms in plants. pp.235–250 In Stratospheric Ozone Reduction, Solar Ultraviolet Radiation and Plant Life. R.C. Worrest and M.M. Caldwell (eds) Springer-Verlag. Berlin, Heidelberg

    Google Scholar 

  • Bennett PV, Hada M, Hidema J, Lepre AM, Pope LC, Quaite FE, Sullivan JH, Takayanagi S, Sutherland JC and Sutherland BM (2001) Isolation of High Molecular Length DNA: Alfalfa, Pea, Rice, Sorghum, Soybean and Spinach. Crop Science 41: 167–172

    Article  CAS  Google Scholar 

  • Björn LO, and Murphy TM (1985) Computer calculation of solar ultraviolet radiation at ground level. Physiologia Vegetatio 23: 555–561

    Google Scholar 

  • Bogenrieder A and Klein R (1982) Does solar UV influence the competitive relationship in higher plants? In Calkins J, (ed.). The role of solar ultraviolet radiation in marine ecosystems. pp.641–649. Plenum Press, New York

    Google Scholar 

  • Buzzell RI, Buttery BR and Bernard RI (1977) Inheritance and linkage of a magenta flower gene in soybeans. Can. J. Genet. Cytology. 19: 749–751

    Google Scholar 

  • Caldwell MM (1971) Solar ultraviolet radiation and the growth and development of higher plants, pp.131–177, In A. C. Giese, ed. Photophysiology, Vol. 6. Academic Press, New York

    Google Scholar 

  • Caldwell MM, Robberecht R and Flint SD (1983) Internal filters: prospects for UV-acclimation in higher plants. Physiol. Plant. 58: 445–450

    Article  CAS  Google Scholar 

  • Caldwell MM, Ballare CL, Bornman JF, Flint SD, Bjorn LO, Teramura AH, Kulandaivelu G, and Tevini M (2003) Terrestrial ecosystems, increased solar ultraviolet radiation and interactions with other climatic change factors. Photochemistry. Photobiology. Science. 2, 29–38

    Article  CAS  Google Scholar 

  • Chappell J and Hahlbrock K (1984) Transcription of plant defense genes in response to UV light or fungal elicitor. Nature (London) 311: 76–78

    Article  CAS  Google Scholar 

  • Cosio EG, and McClure JW (1984) Kaempferol glycosides and enzymes of the flavonol biosynthesis in leaves of a soybean strain with low photosynthetic rates. Plant Physiol. 74, 877–881

    Article  CAS  Google Scholar 

  • Dale JE (1988) The control of leaf expansion. Ann. Rev. Plant Physiol. Plant Molec. Biol. 39: 267–295

    Article  Google Scholar 

  • Dany A-L, Douki T, Triantaphylides C and Cadet J (2001) Repair of the main UV-induced thymine dimeric lesions within Arabidopsis thaliana DNA: evidence for the major involvement of photoreactivation pathways. Journal of Photochemistry and Photobiology 65: 127–135

    Article  CAS  Google Scholar 

  • Day TA (1993) Relating UV-B radiation screening effectiveness of foliage to absorbingcompound concentration and anatomical characteristics in a diverse group of plants. Oecologia 95: 542–550

    Google Scholar 

  • Day TA, Howells BW and Rice WJ (1994) Ultraviolet absorption and epidermal-transmittance spectra in foliage. Physiol. Plant. 92, 207–218

    Article  CAS  Google Scholar 

  • Dillenburg LR, Sullivan JH and Teramura AH (1994) Leaf expansion and development of photosynthetic capacity and pigments in Liquidambar styraciflua—Effects of UV-B radiation. Amer. J. Bot. 82: 878–885w

    Article  Google Scholar 

  • Freeman SE, Blackett AD, Monteleone DC, Setlow RB, Sutherland BM and Sutherland JC (1986) Quantitation of radiation-, chemical-, or enzyme-induced single strand breaks in nonradioactive DNA by alkaline gel electrophoresis: application to pyrimidine dimers. Analytical Biochemistry 158: 119–129

    Article  CAS  Google Scholar 

  • Frohnmeyer H, and Staiger D (2003) Ultraviolet-B Radiation-Mediated Responses in Plants. Balancing Damage and Protection. Plant Physiology 133: 1420–1428

    Article  CAS  Google Scholar 

  • Giese AC (1964) Studies on ultraviolet radiation action upon animal cells. In Photophysiology. Vol 2 (Edited by A.C. Giese), pp.203–245. Academic Press, New York

    Google Scholar 

  • Gitz DC, Liu-Gitz L, Britz SJ and Sullivan JH (2005) Ultraviolet-B effects on stomatal density, water use efficiency and stable carbon isotope discrimination in four glasshouse-grown soybean (Glycine max) cultivars. Env. Exp. Bot. 53: 343–355

    Article  CAS  Google Scholar 

  • Green AES, Cross KR, and Smith LA (1980) Improved Analytical characterization of ultraviolet skylight. Photochemistry Photobiology 31: 59–65

    Article  Google Scholar 

  • Jansen MAK, Gaba V, and Greenberg BM (1998) Higher plants and UV-B radiation: balancing damage, repair and acclimation. Trends in Plant Science 3: 131–135

    Article  Google Scholar 

  • Kang HS, Hidema J, and Kumagai T (1998) Effects of light environment during culture on UV-induced cyclobutyl pyrimidine dimers and their photorepair in rice (Oryza sativa L.). Photochemistry Photobiology 68: 71–77

    CAS  Google Scholar 

  • Kerr RA (1995) At quadrennial geophysics fest, Earth scientists think globally. Science 269: 477–478

    Article  CAS  Google Scholar 

  • Kirk-Davidoff DB, Hintsa EJ, Anderson JD, and Keith DW (1999) The effect of climate change on ozone depletion through changes in stratospheric water vapour. Nature 402: 399–401

    Article  CAS  Google Scholar 

  • Krizek DT, Britz SJ and Mirecki RM (1998) Inhibitory effects of ambient levels of solar UV-A and UV-B radiation on growth of cv. New Red Fire lettuce. Physiologia Plantarum 3: 1–7

    Article  Google Scholar 

  • Liu L and McClure JW (1995) Effects of UV-B on activities of enzymes of secondary phenolic metabolism in barley primary leaves. Physiologia Plantarum 93: 734–739

    Article  CAS  Google Scholar 

  • Liu L, Gitz DC and McClure JW (1995) Effects of UV-B on flavonoids, ferulic acid, growth and photosynthesis in Barley primary leaves. Physiol. Plant. 93: 725–733

    Article  CAS  Google Scholar 

  • Mabry TJ, Markham KR and Thomas MB (1970) The systematic identification of flavonoids. Springer-Verlag, Berlin

    Google Scholar 

  • Mazza CA, Boccalandro H, Giordano CV, Battista D, Scopel AL, and Ballaré CL (2000) Functional significance and induction by solar radiation of ultraviolet-absorbing sunscreens in field-grown soybean crops. Plant Physiology 122: 117–125. Sumere, (eds.). Plenum Press. New York and London. pp. 525–556

    Google Scholar 

  • McClure JW (1986) Physiology of flavonoids in plants. pp. 77–85. In (V. Cody, E. Middleton and J.B. Harborne, eds) Plant Flavonoids in Biology and Medicine: Biochemical, Pharmacological, and Structure-activity Relationships. Alan Risws, Inc.

    Google Scholar 

  • Mohr H and Drumm-Herrel H (1983) Coaction between phytochrome and blue/UV light in anthocyanin synthesis in seedlings. Physiol. Plant. 58: 408–414

    Article  CAS  Google Scholar 

  • Mori T, Nakane M, Hattori T, Matsunaga T, Ihara M, and Nikaido O (1991) Simultaneous establishment of monoclonal antibodies specific for either cyclobutane pyrimidine dimer or (6,4) photoproduct from the same mouse immunized with ultraviolet-irradiated DNA. Photochemistry and Photobiology 54: 225–232

    Article  CAS  Google Scholar 

  • Murali NS and Teramura AH (1986) Effects of supplemental ultraviolet-B radiation on the growth and physiology of field-grown soybean. Environmental and Experimental Botany 26: 233–242

    Article  Google Scholar 

  • Pope LC (2000) Ultraviolet-B radiation effects on two soybean isolines that differ in flavonoid concentration. M.S. Thesis. University of Maryland. 111

    Google Scholar 

  • Quaite FE, Sutherland BM, and Sutherland JC (1992) Quantification of pyrimidine dimers in DNA from UV-B irradiated alfalfa (Medicago sativa L.) seedlings. Applied and Theoretical Electrophoresis 2: 171–175

    CAS  Google Scholar 

  • Quaite FE, Sutherland JC, and Sutherland BM (1994) Isolation of high molecular weight plant DNA for DNA damage quantification: relative effects of solar 297 nm UVB and 365 nm radiation. Plant Molecular Biology 24: 475–483

    Article  CAS  Google Scholar 

  • Raven JA (1977) Evolution of vascular land plants. Adv. Bot. Res. 5: 154–219

    Google Scholar 

  • Robberecht R and Caldwell MM (1978). Leaf epidermal transmittance of ultraviolet radiation and its implications for plant sensitivity to ultraviolet-radiation induced injury. Oecologia (Berl.) 32: 277–287

    Article  Google Scholar 

  • Robberecht R and Caldwell MM (1983) Protective mechanisms and acclimation to solar ultraviolet-B radiation in Oenothera stricta. Plant Cell Environ. 6: 477–485

    Google Scholar 

  • Schnitzler JP, Jungblut TP, Feicht C, Kofferlein M, Langebartels C, Heller W and Sandermann Jr. H (1997) UV-B induction of flavonoid biosynthesis in Scots pine (Pinus sylvestris L.) seedlings. Trees Structure and Function 11: 162–168

    Google Scholar 

  • Statistical Analysis System, Inc. (2000) SAS Users Guide: Statistics. SAS Institute, Inc., Cary, N.C. 584

    Google Scholar 

  • Salawitch RJ, Margitan JJ, Sen B, Toon GC, Osterman GB, Rex M, Elkins JW, Ray EA, Moore FL, Hurst DF, Romashkin PA, Bevilacqua RM, Hoppel KW, Richard EC and Bui TP (2002) Chemical loss of ozone during the Arctic winter of 1999–2000: An analysis based on balloonborne observations, Journal of Geophysical Research 107: 8269

    Article  Google Scholar 

  • Schnitzler JP, Jungblut TP, Feicht C, Kofferlein M, Langebartels C, Heller W and Sandermann Jr. H (1997) UV-B induction of flavonoid biosynthesis in Scots pine (Pinus sylvestris L.) seedlings. Trees Structure and Function 11: 162–168

    Google Scholar 

  • Searles PS, Flint SD and Caldwell MM (2001) A meta-analysis of plant field studies simulating stratospheric ozone depletion. Oecologia 127: 1–10

    Article  Google Scholar 

  • Sheahan JJ (1996) Sinapate esters provide greater UV-B attenuation than flavonoids in Arabidopsis thaliana (Brassicaceae). Amer. J. Bot. 83: 679–686

    Article  CAS  Google Scholar 

  • Sinclair TR, N’Diaya O and Biggs RH (1990) Growth and yield of field-grown soybean in response to enhanced ultraviolet-B radiation. J. Environ Qual. 19: 478–481

    Article  Google Scholar 

  • Stafford HA (1991) Flavonoid evolution: An enzymatic approach. Plant Physiol. 96: 680–685

    Article  CAS  Google Scholar 

  • Stapleton AE (1999) Measurement of UV Radiation-Induced DNA Damage Using Specific Antibodies, In E. D. S. Henderson, ed. DNA Repair Protocols: Eukaryotic Systems, Vol. 113. Humana Press Publishing Agencies, Totowa, NJ

    Google Scholar 

  • Stapleton AE and Walbot V (1994) Flavonoids can protect maize DNA from the induction of ultraviolet radiation damage. Plant Physiology 105: 881–889

    Article  CAS  Google Scholar 

  • Stapleton AE, Mori T, and Walbot V (1993) A simple and sensitive anti-body-based method to measure UV-induced DNA damage in Zea mays. Plant Molecular Biology Reporter 11: 230–236

    Article  CAS  Google Scholar 

  • Stapleton AE, Thorner CS and Walbot V (1997) UV-B component of sunlight causes measurable damage in field-grown maize (Zea mays L.): developmental and cellular heterogeneity of damage and repair. Plant Cell and Environment 20: 279–290

    Article  CAS  Google Scholar 

  • Sullivan JH (2005) Possible impacts of changes in UV-B radiation on North American trees and forests. Invited Review. Env. Pol. 137: 380–389

    CAS  Google Scholar 

  • Sullivan JH and Teramura AH (1989) The effects of ultraviolet-B radiation on loblolly pines I. Growth, photosynthesis and pigment production in greenhouse grown saplings. Physiol. Plant. 77: 202–207

    Article  CAS  Google Scholar 

  • Sullivan JH and Teramura AH (1990) Field studies on the interaction between supplemental ultraviolet-B radiation and drought on soybean growth and physiology. Plant Physiology 92: 141–146

    Article  CAS  Google Scholar 

  • Sullivan JH, Teramura AH and Ziska LH (1992) Variation in UV-B sensitivity in plants from a 3,000 m elevational gradient in Hawaii. Amer. J. Bot. 79: 737–743

    Article  Google Scholar 

  • Sullivan JH, Howells BW, Ruhland C and Day TA (1996) Influence of leaf age and enhanced UV-B radiation on epidermal penetration by UV-B radiation in loblolly pine and sweetgum. Physiologia Plantarum 98: 349–357

    Article  CAS  Google Scholar 

  • Sullivan JH, Gitz DC, Liu-Gitz L, Xu C, Gao W and Slusser J (2007) Coupling short-term changes in ambient UV-B levels with induction of UV-screening compounds. Photochemistry and Photobiology. 83: 863–870

    Article  CAS  Google Scholar 

  • Sutherland BM, Takayanagi S, Sullivan JH, and Sutherland JC (1996) Plant responses to changing environmental stress: cyclobutyl pyrimidine dimer repair in soybean leaves. Photochemistry Photobiology 64: 464–468

    Article  CAS  Google Scholar 

  • Sutherland JC, Monteleone DC, Mugavero JH, and Trunk JG (1987) Unidirectional pulse-field electrophoresis of single-and double-stranded DNA in agarose gels: analytical expressions relating mobility and molecular length and their application in the measurement of strand breaks. Analytical Biochemistry 162: 511–520

    Article  CAS  Google Scholar 

  • Sutherland JC, Chen CZ, Emrick A, Hacham H, Monteleone DC, Ribeiro E, Trunk JG, and Sutherland BM (1990) Lesion measurment in non-radioactive DNA by quantitative gel electrophoresis, pp. 45–61, In Sutherland BM and Woodhead AD (eds). DNA Damage and Repair in Human Tissues. Plenum Press, New York

    Google Scholar 

  • Teramura AH, Sullivan JH, and Lydon J (1990) Effects of UV-B radiation on soybean yield and seed quality: A 6-year field study. Physiologia Plantarum 80: 5–11

    Article  Google Scholar 

  • Tevini M, Braun J and Fieser F (1991) The protective function of the epidermal layer of rye seedlings against ultraviolet-B radiation. Photochem. Photobiol. 53: 329–333

    Article  CAS  Google Scholar 

  • Vogelmann TC (1994) Light within the plant. In: Photomorphogenesis in Plants. 2nd. Ed. (Kendrick RE and Kronenburg GHM, eds.). Kluwer Academic Publishers, Dordecht, The Netherlands. pp. 491–535

    Google Scholar 

  • Waibel AE, Peter T, Carslaw K, Oelhaf H, Wetzel G, Crutzen PJ, Pöschl U, Tsias A, Reimer E, and Fischer H (1999) Arctic ozone loss due to denitrification. Science 283: 2064–2069

    Article  CAS  Google Scholar 

  • Wellman E (1983) UV radiation: Definitions, characteristics and general effects. In Encyclopedia of Plant Physiology, New Series. Vol. 16B. (Shropshire W & Mohr H, ed,), pp.745–756, Springer Verlag, Berlin

    Google Scholar 

  • WMO (World Meteorological Organization) (2003). Scientific Assessment of Ozone Depletion: 2002. Global Ozone Research and Monitoring Project—Report No, 47 Ajavon AN, Albritton DL, Megie G, Watson RT (eds). World Meteorological Organization. Geneva

    Google Scholar 

  • Xu C, Natarajan S and Sullivan JH (2008a) Impact of Solar Ultraviolet-B radiation on the antioxidant defense system in soybean lines differing in flavonoids biosynthesis. Environmental and Experimental Botany 63: 39–48

    Article  CAS  Google Scholar 

  • Xu C, Sullivan JH, Garrett WM, Caperna TJ, Natarajan S (2008b) Impact of solar ultraviolet-B radiation on the proteome in soybean lines differing in flavonoid contents. Phytochemistry 69: 38–48

    Article  CAS  Google Scholar 

  • Ziska LE, Teramura AH and Sullivan JH (1992) Physiological sensitivity to plants along an elevational gradient to UV-B radiation. American Journal of Botany 79: 863–871

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Plant Science and Landscape Architecture, 2122 Plant Sciences Building University of Maryland, College Park, MD, 20742, USA

    Joseph H. Sullivan & Linda C. Pope

  2. Brookhaven National Laboratory, P.O. Box 5000, Upton, NY, 11973-5000, USA

    Betsy M. Sutherland & Paula V. Bennett

  3. Department of Mathematics and Statistics, University of North Carolina at Wilmington, Wilmington, NC, 28403, USA

    James E. Blum

  4. Department of Biology and Marine Biology, University of North Carolina at Wilmington, Wilmington, NC, 28403, USA

    Ann E. Stapleton

  5. USDA-ARS, 3810 4th Street, Lubbock, Texas, 79415, USA

    Dennis C. Gitz III

Authors
  1. Joseph H. Sullivan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Linda C. Pope
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Betsy M. Sutherland
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Paula V. Bennett
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. James E. Blum
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ann E. Stapleton
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Dennis C. Gitz III
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Natural Resource Ecology Laboratory, Colorado State University, USA

    Wei Gao  & James R. Slusser  & 

  2. Cooperative State Research Education & Extension Service, Waterfront Centre, Ste.3422 800 9th Street SW, Washington DC, 20024, USA

    Daniel L. Schmoldt

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Tsinghua University Press, Beijing and Springer-Verlag Berlin Heidelberg

About this chapter

Cite this chapter

Sullivan, J.H. et al. (2010). Assessment of DNA Damage as a Tool to Measure UV-B Tolerance in Soybean Lines Differing in Foliar Flavonoid Composition. In: Gao, W., Slusser, J.R., Schmoldt, D.L. (eds) UV Radiation in Global Climate Change. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-03313-1_15

Download citation

Publish with us

Policies and ethics

Search

Navigation