Abstract
We investigated the radiation damages caused by two types of proton beam and gamma ray in the Cymbidium hybrid RB001 [(C. sinensis × C. goeringii) × Cymbidium spp.] to characterize proton beam as a new mutagen for Cymbidium mutation breeding. The protocorm-like bodies (PLBs) of Cymbidium hybrid were irradiated with a 45 MeV proton beam [mean linear energy transfer (LET) = 1.461 keV·μm−1], a 100 MeV proton beam (LET = 0.7306 keV·μm−1), and gamma ray (LET = 0.2 keV·μm−1). The PLBs treated with radiation doses of 0–100 Gy were analyzed using the comet assay and their physiological responses as indices of radiation damage. In the comet assay, the 45 MeV proton beam caused significant damage to the DNA integrity, but the 100 MeV proton beam and gamma ray showed relatively little radiation damage between the untreated control and treated PLBs. Malondialdehyde (MDA) content, an index of the indirect effects of ionizing radiation, was increased slightly by the 45 MeV proton beam with higher-LET, but highly increased by the 100 MeV proton beam with lower-LET. These results suggested that the 100 MeV proton beam caused extreme oxidative stress. Therefore, proton beam have unique characteristics in DNA mutagenesis pattern according to its LETs. Based on these results, proton beam is expected to be a useful tool for developing new mutant varieties of Cymbidium.
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Literature Cited
Boudaïffa, B., P. Cloutier, D. Hunting, M.A. Huels, and L. Sanche. 2000. Resonant formation of DNA strand breaks by low-energy (3 to 20 eV) electrons. Science 287:1658–1660.
Del Rio, D., A.J. Stewart, and N. Pellegrini. 2005. A review of recent studies on malondialdehyde as toxic molecule and biological marker of oxidative stress. Nutr. Metabol. Cardiovasc. Dis. 15:316–328.
Dhawan, A., M.M. Bajpayee, A.K. Pandey, and D. Parmar. 2009. Protocol for the single cell gel electrophoresis/comet assay for rapid genotoxicity assessment. Sigma 1077:1–10.
Du Puy, D., P. Cribb, M. Tibbs, and K. Royal Botanic Gardens. 2007. The genus Cymbidium. Timber Press. Portland, OR.
Gedik, C., W. Ewen, and A. Collins. 1992. Single-cell gel electrophoresis applied to the analysis of UV-C damage and its repair in human cells. Intl. J. Radiat. Biol. 62:313–320.
Gichner, T., Z. Patková, J. Száková, and K. Demnerová. 2004. Cadmium induces DNA damage in tobacco roots, but no DNA damage, somatic mutations or homologous recombination in tobacco leaves. Mutat. Res. 559:49–57.
Gichner, T., Z. Patková, J. Száková, and K. Demnerová. 2006. Toxicity and DNA damage in tobacco and potato plants growing on soil polluted with heavy metals. Ecotoxicol. Environ. Saf. 65:420–426.
Gichner, T., Z. Patková, J. Száková, I. Žnidar, and A. Mukherjee. 2008. DNA damage in potato plants induced by cadmium, ethyl methanesulphonate and γ-rays. Environ. Exp. Bot. 62:113–119.
Hase, Y., M. Yamaguchi, M. Inoue, and A. Tanaka. 2002. Reduction of survival and induction of chromosome aberrations in tobacco irradiated by carbon ions with different linear energy transfers. Intl. J. Radiat. Biol. 78:799–806.
Hase, Y., Y. Akita, S. Kitamura, I. Narumi, and A. Tanaka. 2012. Development of an efficient mutagenesis technique using ion beams: Toward more controlled mutation breeding. Plant Biotechnol. 29:193–200.
Hirono, Y, H.H. Smith, J.T. Lyman, K.H. Thompson, and J.W. Baum. 1970. Relative biological effectiveness of heavy ions in producing mutations, tumors, and growth inhibition in the crucifer plant, Arabidopsis. Radiat. Res. 44: 204–223.
Joint FAO/IAEA Programme. 2014. Mutant Variety Database (MVD). http://mvgs.iaea.org/AboutMutantVarities.aspx.
Kahl, G. and K. Meksem. 2010. The handbook of plant mutation screening: Mining of natural and induced alleles. Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany.
Kanaya, T., H. Saito, Y. Hayashi, N. Fukunishi, H. Ryuto, K. Miyazaki, T. Kusumi, T. Abe, and K. Suzuki. 2008. Heavy-ion beam-induced sterile mutants of verbena (Verbena × hybrida) with an improved flowering habit. Plant Biotechnol. 25:91–96.
Kazama, Y., H. Saito, Y. Yamamoto, Y. Hayashi, H. Ichida, H. Ryuto, N. Fukunishi, and T. Abe. 2008. LET-dependent effects of heavy-ion beam irradiation in Arabidopsis thaliana. Plant Biotechnol. 25:113–117.
Khan, M.H. and S.K. Panda. 2008. Alterations in root lipid peroxidation and antioxidative responses in two rice cultivars under NaCl-salinity stress. Acta Physiol. Plant. 30:81–89.
Kikuchi, O.K. 2000. Orchid flowers tolerance to gamma-radiation. Radiat. Phys. Chem. 57:555–557.
Kim, S.K., H.J. Choi, K.R. Kim, and H.Y. Kim. 2011a. Properties of starches in Chinese yam, Dioscorea oppsita Thunb. Irradiated with Proton Beam. Kor. J. Plant. Res. 24:304–308.
Kim, S.K., H.J. Choi, K.R. Kim, I.J. Lee, and H.Y. Kim. 2011b. Effect of proton beam radiation on bulbil yield and gibberellins of Chinese yam (Dioscorea opposita Thunb.). Kor. J. Crop Sci. 56:250–254.
Kim, S.K and H.Y. Kim. 2013. Effect of high proton beam irradiation on pasting properties of rice starch. J. Crop Sci. Biotech. 16: 161–166.
Kim, S.K., S.Y. Park, and H.Y. Kim. 2013. Characterization of physicochemical properties of starch in barley irradiated with proton beam. Kor. J. Crop Sci. 58:260–266.
Kumar, B., S. Kumar, and M. Thakur. 2012. In vitro mutation induction and selection of chrysanthemum (Dendranthema grandiflora Tzelev) lines with improved resistance to Septoria obesa Syd. Intl. J. Plant Res. 2:103–107.
Larson, R.A. 1988. The antioxidants of higher plants. Phytochemistry 27:969–978.
Liman, R., ].H. Ciğerci, D. Akyıl, Y. Eren, and M. Konuk. 2011. Determination of genotoxicity of fenaminosulf by Allium and comet tests. Pestic. Biochem. Physiol. 99:61–64.
Luan, L.Q., N.H.P. Uyen, and V.T.T. Ha. 2012. In vitro mutation breeding of Paphiopedilum by ionization radiation. Sci. Hortic. 144:1–9.
Mehnati, P., S. Morimoto, F. Yatagai, Y. Furusawa, Y. Kobayashi, S. Wada, T. Kanai, F. Hanaoka, and H. Sasaki. 2005. Exploration of ‘Over Kill Effect’ of high-LET Ar- and Fe-ions by evaluating the fraction of non-hit cell and interphase death. J. Radiat. Res. 46:343–350.
Menke, M., K.J. Angelis, and I. Schubert. 2000. Detection of specific DNA lesions by a combination of comet assay and FISH in plants. Environ. Mol. Mutagen. 35:132–138.
Ministry of Agriculture, Food and Rural Affairs. 2012. Flower cultivation status. http://library.mafra.go.kr/skyblueimage/15939.pdf.
Nagatomi, S. 2003. Development of flower mutation breeding through ion beam irradiation. Res. J. Food Agr. 26:33–38.
Okamura, M., N. Yasuno, M. Ohtsuka, A. Tanaka, N. Shikazono, and Y. Hase. 2003. Wide variety of flower-color and-shape mutants regenerated from leaf cultures irradiated with ion beams. Nucl. Instrum. Meth. Phys. Res. 206:574–578.
Olive, P.L and J.P. Banáth. 2006. The comet assay: A method to measure DNA damage in individual cells. Nat. Protocol 1:23–29.
Puchooa, D. 2005. In vitro mutation breeding of Anthurium by gamma radiation. Intl. J. Agr. Biol. 7:11–20.
Schulz-Ertner, D. and H. Tsujii. 2007. Particle radiation therapy using proton and heavier ion beams. J. Clin. Oncol. 25:953–964.
Shikazono, N., C. Suzuki, S. Kitamura, H. Watanabe, S. Tano, and A. Tanaka. 2005. Analysis of mutations induced by carbon ions in Arabidopsis thaliana. J. Exp. Bot. 56:587–596.
Shikazono, N., Y. Yokota, A. Tanaka, H. Watanabe, and S. Tano. 1998. Molecular analysis of carbon ion-induced mutations in Arabidopsis thaliana. Genes Genet. Syst. 73:173–179.
Shu, Q.Y., B.P. Forster, and H. Nakagawa. 2012. Plant mutation breeding and biotechnology. Co-published with the Food and Agriculture Organization of the United Nations (FAO) and International Atomic Energy Agency (IAEA).
Tanaka, A., N. Shikazono, and Y. Hase. 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. Radiat. Res. 51:223–233.
Tanaka, A., N. Shikazono, Y. Yokota, H. Watanabe, and S. Tano. 1997. Effects of heavy ions on the germination and survival of Arabidopsis thaliana. Intl. J. Radiat. Biol. 72:121–127.
Unyayar, S., A. Celik, F.O. Cekic, and A. Gozel. 2006. Cadmiuminduced genotoxicity, cytotoxicity and lipid peroxidation in Allium sativum and Vicia faba. Mutagenesis 21:77–81.
Wang, Y.C., G.Z. Qu, H.Y. Li, Y.J. Wu, C. Wang, G.F. Liu, and C.P. Yang. 2010. Enhanced salt tolerance of transgenic poplar plants expressing a manganese superoxide dismutase from Tamarix androssowii. Mol. Biol. Rep. 37:1119–1124.
Yamaguchi, H., A. Shimizu, K. Degi, and T. Morishita. 2008. Effects of dose and dose rate of gamma ray irradiation on mutation induction and nuclear DNA content in chrysanthemum. Breed. Sci. 58: 331–335.
Yokota, Y., Y. Hase, N. Shikazono, A. Tanaka, and M. Inoue. 2003. LET dependence of lethality of carbon ion irradiation to single tobacco cells. Intl. J. Radiat. Biol. 79:681–685.
Yokota, Y., S. Yamada, Y. Hase, N. Shikazono, I. Narumi, A. Tanaka, and M. Inoue. 2007. Initial yields of DNA double-strand breaks and DNA fragmentation patterns depend on linear energy transfer in tobacco BY-2 protoplasts irradiated with helium, carbon and neon ions. Radiat. Res. 167:94–101.
Zaka, R., C.M. Vandecasteele, and M.T. Misset. 2002. Effects of low chronic doses of ionizing radiation on antioxidant enzymes and G6PDH activities in Stipa capillata (Poaceae). J. Exp. Bot. 53:1979–1987.
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Lee, YM., Jo, Y.D., Lee, HJ. et al. DNA damage and oxidative stress induced by proton beam in Cymbidium hybrid. Hortic. Environ. Biotechnol. 56, 240–246 (2015). https://doi.org/10.1007/s13580-015-0081-6
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DOI: https://doi.org/10.1007/s13580-015-0081-6