ISHS
  eJHS
     
EJHS
Home


Submit
an article


Subscriptions

ISHS Home

ISHS Contact

Search

eJHS
  Eur.J.Hortic.Sci. 81 (6) 303-309 | DOI: 10.17660/eJHS.2016/81.6.3
ISSN 1611-4426 print and 1611-4434 online | © ISHS 2016 | European Journal of Horticultural Science | Original article

Oxidative stress and DNA damage induced by gamma irradiation in Korean lawngrass (Zoysia japonica Steud.)

Hyo-Jeong Lee1,†, Ye-Sol Kim2,†, Yeong Deuk Jo2, Bo-Keun Ha3, Dong Sub Kim2, Jin-Baek Kim2, Si-Yong Kang2 and Sang Hoon Kim2
1Crop Breeding Division, National Institute of Crop Science, RDA, Wanju 55365, Korea
2Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 56212, Korea
3Division of Plant Biotechnology, College of Agriculture and Life Sciences, Chonnam National University, Gwangju 61186, Korea
†These authors contributed equally to this work.

SUMMARY
Korean lawngrass is a widely-used turfgrass species. In this study, we determined the effects of gamma irradiation on growth characteristics and cellular damages in this species. Seeds were irradiated with 100, 200, 400, 600, 800, and 1,000 Gy of gamma rays, respectively. To evaluate growth characteristics, germination rate and morphological traits (shoot length and width, root length, fresh weight, leaf blade length and width, sheath length, plant width, and number of tillers) were measured. We also examined the activities of the antioxidant enzymes including ascorbate peroxidase, catalase, superoxide dismutase, and peroxidase and the content of malondialdehyde. The germination rate decreased with an increase of gamma-ray dose, and seeds irradiated with doses higher than 600 Gy did not germinate at all. The 50% lethal dose (LD50) for gamma irradiation was 280-300 Gy. We also conducted a comet assay to observe nuclear DNA damage in the germinated plants. Significant differences were identified between the control and 400 Gy treatments. Increasing gamma-ray doses from 0 to 400 Gy, the percentage of head DNA decreased significantly from 93.9% to 67.0%. In addition, the antioxidant enzyme activities and the content of MDA increased with the gamma-ray doses. These results showed that higher doses of gamma irradiation caused the greater oxidative stress and DNA damage in Korean lawngrass.

Keywords gamma-ray, antioxidant enzyme, comet assay

Significance of this study

What is already known on this subject?

  • According to heat tolerance and winter hardiness, Korean lawngrass is one of the important ground cover plants in Asia, USA, and other countries. Gamma ray have been used to induce mutants with various trait in plants. However, gamma ray also induce oxidative stress with the overproduction of reactive oxygen species (ROS), superoxide radicals (O2-), hydroxyl radicals (•OH), and hydrogen peroxide (H2O2). Furthermore, comet assay detected DNA damage on nucleus.
What are the new findings?
  • Germination rate and morphological traits according to plant development reduced with increasing dose of gamma ray. Thus, DNA damage of irradiation was detected at increasing dose rates with 100 Gy and higher. Significant difference was not found in activities of oxidative stress related enzymes between gamma-irradiation dose rate. But MDA content increased with the gamma-ray doses. Thus, higher doses of gamma irradiation induced the oxidative stresses and DNA damages in Korean lawngrass.
What is the expected impact on horticulture?
  • According to these results, gamma rays affected morphological traits, lipid peroxidation, and DNA damage in Korean lawngrass. These results will help further research of mutation breeding using irradiation and provide a guidance of selection of dose and dose rate for induction of various traits.

Download fulltext version How to cite this article       Export citation to RIS format      

E-mail: shkim80@kaeri.re.kr  

References

  • Achary, V.M.M., Jena, S., Panda, K.K., and Panda, B.B. (2008). Aluminium induced oxidative stress and DNA damage in root cells of Allium cepa L. Ecotoxicol. and Environ. Safety 70, 300–310. https:/doi.org/10.1016/j.ecoenv.2007.10.022.

  • Aebi, H. (1984). Catalase in vitro. Methods in Enzymol. 105, 121–126. https:/doi.org/10.1016/S0076-6879(84)05016-3.

  • Al-Rumaih, M.M., and Al-Rumaih, M.M. (2008). Influence of ionizing radiation on antioxidant enzymes in three species of Trionella. Amer. J. Environ. Sci. 4, 151–156. https:/doi.org/10.3844/ajessp.2008.151.156.

  • Apel, K., and Hirt, H. (2004). Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu. Rev. Plant Biol. 55, 373–399. https:/doi.org/10.1146/annurev.arplant.55.031903.141701.

  • Beyer Jr., W.F., and Fridovich, I. (1987). Assaying for superoxide dismutase activity: some large consequences of minor changes in conditions. Anal. Biochem. 161, 559–566. https:/doi.org/10.1016/0003-2697(87)90489-1.

  • Bowler, C., Montagu, M.V., and Inze, D. (1992). Superoxide dismutase and stress tolerance. Ann. Rev. of Plant Biol. 43, 83–116. https:/doi.org/10.1146/annurev.pp.43.060192.000503.

  • Bradford, M. (1976). A rapid and sensitive method for quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254. https:/doi.org/10.1016/0003-2697(76)90527-3.

  • Chance, B., and Maehly, A.C. (1955). Assay of catalase and peroxidases. Method Enzymol. 2, 764–775. https:/doi.org/10.1016/S0076-6879(55)02300-8.

  • Collins, A.R. (2014), Measuring oxidative damage to DNA and its repair with the comet assay. Biochimica et Biophysica Acta (BBA) - General Subjects 1840, 794–800. https:/doi.org/10.1016/j.bbagen.2013.04.022.

  • Datta, S.K. (2009). A Report on 36 years of practical work on crop improvement through induced mutagenesis: Induced plant mutations in the genomics era. In Joint FAO/IAEA Division of Nuclear Techniques in Food and Agricultures, International Atomic Energy Agency, Q.Y. Shu, ed. (Vienna, Austria), p.253–256.

  • Dhawan, A., Bajpayee, M., Pandey, A.K., and Parmar, D. (2009). Protocol for the single cell gel electrophoresis/Comet assay for rapid genotoxicity assessment. ITRC: The SCGE/Comet assay protocol (Lucknow: Industrial Toxicology Research Centre), p.1–10.

  • Dhindsa, R.S., and Matowe, W. (1981). Drought tolerance in two mosses correlated with enzymatic defense against lipid peroxidation. J. Expt. Bot. 32, 79–91. https:/doi.org/10.1093/jxb/32.1.79.

  • Egert, M., and Tevini, M. (2002). Influence of drought on some physiological parameters symptomatic for oxidative stress in leaves of chives (Allium schoenoprasum). Environ. Expt. Bot. 48, 43–49. https:/doi.org/10.1016/S0098-8472(02)00008-4.

  • Georgieva, M., and Stoilov, L. (2008). Assessment of DNA strand breaks induced by bleomycin in barley by the comet assay. Environ. Mol. Mutagenesis 49, 381–387. https:/doi.org/10.1002/em.20396.

  • IAEA (2014). Joint FAO/IAEA Mutation Varieties Database. http://www.mvd.iaea.org.

  • Kim, D.S., Lee, I.S., Jang, C.S., Kang, S.Y., Park, I.S., Song, H.S., and Seo, Y.W. (2005). High amino acid accumulating 5-methyltryptophan-resistant rice mutants may include an increased antioxidative response system. Physiologia Plantarum 123, 302–313. https:/doi.org/10.1111/j.1399-3054.2005.00468.x.

  • Kim, D.S., Song, M.R., Kim, S.H., Jang, D.S., Kang, S.Y., Kim, J.B., Kim, S.H., and Ha, B.G. (2011). Physiological responses of rice by acute and chronic gamma irradiation. J. Radiat. Industry 5, 55–62.

  • Kim, J.S., Lee, Y.K., Park, H.S., Back, M.H., and Kim, D.H. (2000). Influence of low dose gamma radiation on the growth of maize (Zea mays L.) varieties. Kor. J. Environ. Agr. 19, 328–331.

  • Koppen, G., Toncelli, L.M., Triest, L., and Verschaeve, L. (1999). The comet assay: a tool to study alteration of DNA integrity in developing plant leaves. Mechanisms of Ageing and Dev. 110, 13–24. https:/doi.org/10.1016/S0047-6374(99)00038-X.

  • Kristina, G., Shirshankar, G., and Nath, J. (1984). Non-arrowing and broad-leaved mutants in Rhodes grass (Chloris gayana Kunth.). Euphytica 33, 525–528. https:/doi.org/10.1007/BF00021153.

  • Lagoda, P.J.L. (2012). Effect of radiation on living cells and plants. In Plant Mutation Breeding and Biotechnology, Q.Y. Shu, B.P. Forster, and H. Nakagawa, eds. (Cambridge, Mass.: CABI), p.58–59. https:/doi.org/10.1079/9781780640853.0123.

  • Lee, H.J., Lee, G.J., Kim, D.S., Kim, J.B., Ku, J.H., and Kang, S.Y. (2008). Determination of the optimum dose range for a mutation induction of turfgrass by a gamma-ray. Kor. Turfgrass Sci. 22, 25–34.

  • Li, R., Bruneau, A.H., and Qu, R. (2010). Morphological mutants of St. Augustinegrass induced by gamma ray irradiation. Plant Breeding 129, 412–416. https:/doi.org/10.1111/j.1439-0523.2009.01735.x.

  • Lu, S., Wang, Z., Niu, Y., Chen, Y., Chen, H., Fan, Z., Lin, J., Yan, K., Guo, Z., and Li, H. (2009). Gamma-ray radiation induced dwarf mutants of turf-type bermudagrass. Plant Breeding 128, 205–209. https:/doi.org/10.1111/j.1439-0523.2008.01544.x.

  • Lutts, S., Kinet, J.M., and Bouharmont, J. (1996). NaCl-induced senescence in leaves of rice (Oryza sativa L.) cultivars differing in salinity resistance. Ann. Bot. 78, 389–398. https:/doi.org/10.1006/anbo.1996.0134.

  • Masche, R., Lippmann, B., Holzinger, S., and Bergmann, H. (2002). Arsenate toxicity: effects on oxidative stress response molecules and enzymes in red clover plants. Plant Sci. 163, 961–969. https:/doi.org/10.1016/S0168-9452(02)00245-5.

  • Merten, M., Chen, I.P., Angelis, K.J., and Ingo, S. (2001). DNA damage and repair in Arabidopsis thaliana as measured by the comet assay after treatment with different classes of genotoxins. Mutation Research/Genetic Toxicology and Environmental Mutagenesis 493, 87–93. https:/doi.org/10.1016/S1383-5718(01)00165-6.

  • Mittler, R., and Zilinskas, B.A. (1993). Detection of ascorbate peroxidase activity in native gels by inhibition of ascorbate dependent reduction of nitroblue tetrazolium. Analytical Biochemistry 112, 540–546. https:/doi.org/10.1006/abio.1993.1366.

  • Navarrete, M.H., Carrera, P., De Miguel, M., and De La Torre, C. (1997). A fast comet assay variant for solid tissue cells. The assessment of DNA damage in higher plants. Mutation Research/Genetic Toxicology and Environmental Mutagenesis 389, 271–277. https:/doi.org/10.1016/S1383-5718(96)00157-X.

  • Nemavarkar, P.S., Chourasia, B.K., and Pasupathy, K. (2004). Detection of γ-irradiation induced DNA damage and radioprotection of compounds in yeast using comet assay. J. Radiat. Res. 45, 169–174. https:/doi.org/10.1269/jrr.45.169.

  • Noreen, Z., and Ashraf, M. (2009). Assessment of variation in antioxidative defense system in salt-treated pea (Pisum sativum) cultivars and its putative use as salinity tolerance markers. J. Plant Physiol. 166, 1764–1774. https:/doi.org/10.1016/j.jplph.2009.05.005.

  • Pόtter, J. (1974). Peroxidase. In Methods of Enzymatic Analysis, H.U. Bergmeyer, ed. (Weinheim, USA: Verlag Chemie-Academic Press), p.685–690. https:/doi.org/10.1016/B978-0-12-091302-2.50033-5.

  • Scandalios, J.G. (1993). Oxygen stress and superoxide dismutase. Plant Physiol. 101, 7–12. https:/doi.org/10.1104/pp.101.1.7.

  • Scebba, F., Sebastiani, L., and Vitagliano, C. (1999). Protective enzyme against activated oxygen species in wheat (Triticum aestivum L.) seedlings: Responses to cold acclimation. J. of Plant Physiol. 155, 762–768. https:/doi.org/10.1016/S0176-1617(99)80094-7.

  • Stavreva, D.A., and Gichner, T. (2002). DNA damage induced by hydrogen peroxide in cultured tobacco cells is dependent on the cell growth stage. Mutation Research/Genetic Toxicology and Environmental Mutagenesis 514, 147–152. https:/doi.org/10.1016/S1383-5718(01)00330-8.

  • Tanaka, A., Shikazono, N., and Hase, Y. (2010). Studies on biological effects of ion beams on lethality, molecular nature of mutation, mutation rate, and spectrum of mutation phenotype for mutation breeding in higher plants. J. of Radiation Research 51, 223–233. https:/doi.org/10.1269/jrr.09143.

  • Toyama, K., Bae, C.H., Kang, J.G., Lim, Y.P., Adachi, T., Riu, K.Z., Song, P.S., and Lee, H.Y. (2003). Production of herbicide-tolerant zoysiagrass by Agrobacterium-mediated transformation. Mol. Cell 16, 19–27.

  • Wang, Y., Jiang, J., Zhao, X., Liu, G., Yang, C., and Zhan, L. (2006). A novel LEA gene from Tamarix androssowii confers drought tolerance in transgenic tobacco. Plant Sci. 171, 655–662. https:/doi.org/10.1016/j.plantsci.2006.06.011.

  • White, R.H., Engelke, M.C., Anderson, S.J., Ruemmele, B.A., Marcum, K.B., and Taylor, G.R. (2001). Zoysiagrass water relations. Crop Sci. 41, 133–138. https:/doi.org/10.2135/cropsci2001.411133x.

  • Wu, D., and Cederbaum, A.I. (2003). Alcohol, oxidative stress, and free radical damage. Alcohol Research and Health 27, 277–284.

Received: 15 April 2016 | Accepted: 28 October 2016 | Published: 23 December 2016 | Available online: 23 December 2016

previous article     Volume 81 issue 6     next article