Ye-Sol Kim¹, Sang Yeop Sung¹, Yeong Deuk Jo¹, Hyo-Jeong Lee² and Sang Hoon Kim^1
1Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 56212, Korea
²Crop Breeding Division, National Institute of Crop Science, RDA, Wanju 55365, Korea

SUMMARY Gamma rays are useful to develop new cultivars for mutation breeding, in which the total dose and dose rate are important. On two chrysanthemum cultivars, 'Noble Wine' and 'Pinky', were tested the effect of the dose rate. Four weeks after irradiation, their survival rate and plant height were significantly decreased at the dose rate 30 Gy/1 h. The dose rate 30 Gy/4 h produced the highest mutation frequency in both cultivars, with a wide mutation spectrum. Moreover, sucrose is known as stimulus of anthocyanin biosynthesis. For the induction of anthocyanin related flower-color mutants, we pre-treated 4-week old plants with 100, 200 and 300 mM sucrose before gamma irradiation. The cultivar 'Pinky' showed a high survival rate in all treatments; while almost all 'Noble Wine' plants treated with 200 and 300 mM sucrose died. A number of mutated branches were identified at 100 mM sucrose + 30 Gy/24 h of irradiation in the cultivar 'Noble Wine'. Thus, a proper dose rate of gamma rays and sucrose pre-treatment could induce flower color mutation in chrysanthemum. Keywords anthocyanin biosynthesis, mutation breeding, mutation frequency

Significance of this study What is already known on this subject? Mutation breeding is widely used to develop new cultivars including flower color mutants. Especially gamma rays are one of the important mutagens. To determine proper dose and dose rate is important to induce mutation using gamma rays. Furthermore, sucrose is a stimulator of anthocyanin biosynthesis. When target genes are highly expressed, irradiation is more effective. What are the new findings? Survival rate and plant height were decreased with increasing dose rate, particularly in the cultivar 'Pinky'. Plants irradiated with 30 Gy for 4 h showed a high mutation frequency with a wide color spectrum in both cultivars. In addition, irradiated plants with 100 mM sucrose pre-treatment induced many flower color mutants however, most all plant wither on 200 mM and 300 mM of sucrose treatment in the cultivar 'Noble Wine'. What is the expected impact on horticulture? Based on these results, it will help research suitable dose rate in gamma ray mutagenesis with various mutation spectrum in chrysanthemum. A combination of sucrose pre-treatment and gamma irradiation is effective to induce mutants. It would encourage to develop new cultivars using gamma rays.

E-mail: shkim80@kaeri.re.kr  

References

• Bottino, P.J., Sparrow, A.H., Schwemmer, S.S., and Thompson, K.H. (1975). Interrelation of exposure and exposure rate in germinating seeds of barley and its concurrence with dose-rate theory. Rad. Bot. 15, 17–27. https://doi.org/10.1016/S0033-7560(75)80010-X.

• Broertjes, C. (1968). Dose-rate effects in Saintpaulia. In Mutations in plant breeding II, Proceedings of a panel, jointly organized by the IAEA and FAO, IAEA. (Vienna, Austria), pp. 63–71.

• Broertjes, C., and Van Harten, A.M. (1988). Applied mutation breeding for vegetatively propagated crops (Amsterdam: Elsevier), pp. 85–86.

• Broertjes, C., Koene, P., and Van Veen, J.W.H. (1980). A mutant of a mutant of a…: Irradiation of progressive radiation-induced mutants in a mutation-breeding programme with Chrysanthemum morifolium Ram. Euphytica 29, 525–530. https://doi.org/10.1007/BF00023198.

• Datta, S.K., Misra, P., and Mandal, A.K.A. (2005). In vitro mutagenesis – a quick method for establishment of solid mutant in chrysanthemum. Curr. Sci. 88, 155–158.

• De Jong, J., and Custers, J.B.M. (1986). Induced changes in growth and flowering of chrysanthemum after irradiation and in vitro culture of pedicels and petal epidermis. Euphytica 35, 137–148. https://doi.org/10.1007/BF00028551.

• Gollop, R., Even, S., Colova-Tsolova, V., and Peri, A. (2002). Expression of the grape dihydroflavonol reductase gene and analysis of its promoter region. J. Exp. Bot. 53, 1397–1409. https://doi.org/10.1093/jexbot/53.373.1397.

• Gollop, R., Farhi, S., and Peri, A. (2001). Regulation of the leucoanthcyanidin dioxygenase gene expression in Vitis vinifera. Plant Sci. 161, 579–588. https://doi.org/10.1016/S0168-9452(01)00445-9.

• Goo, D.H., Yae, B.W., Song, H.S., Park, I.S., Han, B.H., and Yu, H.J. (2003). Color change in chrysanthemum flower by gamma ray irradiation. J. Kor. Soc. Hort. Sci. 44, 1006–1009.

• Hara, M., Oki, K., Hoshino, K., and Kuboi, T. (2003). Enhancement of anthocyanin biosynthesis by sugars in radish (Raphanus sativus) hypocotyl. Plant Sci. 164, 259–265. https://doi.org/10.1016/S0168-9452(02)00408-9.

• Hase, Y., Okamura, M., Takeshita, D., Narumi, I., and Tanaka, A. (2010). Efficient induction of flower-color mutants by ion beam irradiation in petunia seedlings treated with high sucrose concentration. Plant Biotechnol. 27, 99–103. https://doi.org/10.5511/plantbiotechnology.27.99.

• Hase, Y., Akita, Y., Kitamura, S., Narumi, I., and Tanaka, A. (2012). Development of an efficient mutagenesis technique using ion beams: Toward more controlled mutation breeding. Plant Biotechnol. 29, 193–200. https://doi.org/10.5511/plantbiotechnology.12.0106a.

• Hwang, J.C., Chin, Y.D., Chung, Y.M., and Kim, S.G. (2008). A new early flowering, spray chrysanthemum cultivar for cut flower, ‘Pinky’ with single type and pink petals. Korean J. Breed. Sci. 40, 196–200.

• Hwang, J.C., Chin, Y.D., Chung, Y.M., Kim, S.G., and Jeong, B.Y. (2009). A new spray chrysanthemum cultivar, ‘Noble Wine’ with good color, single type and bi-color petals for cut flower. Flower Res. J. 17, 137–140.

• Kang, E.J., Lee, Y.M., Sung, S.Y., Ha, B.K., Kim, S.H., Kim, D.S., Kim, J.B., and Kang, S.Y. (2013). Analysis of the genetic relationship of gamma-irradiated in vitro mutants derived from standard-type chrysanthemum cv. Migok. Hort. Environ. Biotechnol. 54, 76–81. https://doi.org/10.1007/s13580-013-0124-9.

• Killion, D.D., and Constantin, M.J. (1971). Acute gamma irradiation of the wheat plant: effects of exposure, exposure rate, and developmental stage on survival, height, and grain yield. Rad. Bot. 11, 367–373. https://doi.org/10.1016/S0033-7560(71)90850-7.

• Kim, S.H., Chung, S.J., Lee, G.J., Kim, D.S., Kim, J.B., and Kang, S.Y. (2009). Mutation breeding of various spray chrysanthemum cultivars by gamma-ray irradiation J. Rad. Ind. 3, 227–232.

• Lamseejan, S., Jompuk, P., Wongpiyasatid, A., Deeseepan, S., and Kwanthammachart, P. (2000). Gamma-rays induced morphological changes in chrysanthemum (Chrysanthemum morifolium). Kasetsart J. (Nat. Sci.) 34, 417–422

• Loreti, E., Alpi, A., and Perata, P. (2000). Glucose and disaccharide-sensing mechanisms modulate the expression of α-amylase in barley embryos. Plant Physiol. 123, 939–948. https://doi.org/10.1104/pp.123.3.939.

• Mabuchi, T., and Matsumura, S. (1964). Dose rate dependence of mutation rates from ɣ-irradiated pollen grains of maize. Jap. J. Genet. 39, 131–135. https://doi.org/10.1266/jjg.39.131.

• Misra, P., and Datta, S.K. (2007). Standardization of in vitro protocol in Chrysanthemum cv. Madam E. Roger for development of quality planting material and to induce genetic variability using gamma-radiation. Ind. J. Biotech. 6, 121–124.

• Misra, P., Datta, S.K., and Chakrabarty, D. (2003). Mutation in flower colour and shape of Chrysanthemum morifolium induced by gamma-radiation. Biol. Plant. 47, 153–156. https://doi.org/10.1023/A:1027365822769.

• Mita, S., Murano, N., Akaike, M., and Nakamura, K. (1997). Mutants of Arabidopsis thaliana with pleiotropic effects on the expression of the gene for β-amylase and on the accumulation of anthocyanin that are inducible by sugars. Plant J. 11, 841–851. https://doi.org/10.1046/j.1365-313X.1997.11040841.x.

• Nagatomi, S., Miyahira, E., and Degi, K. (1997). Combined effect of gamma irradiation methods and in vitro explants sources on mutation induction of flower color in Chrysanthemum morifolium Ramat. Gamma Field Symp. 35, 51–69.

• Nauman, C.H., Underbrick, A.G., and Sparrow, A.H. (1975). Influence of radiation dose rate on somatic mutations induction in Tradescantia stamen hairs. Rad. Res. 62, 79–96. https://doi.org/10.2307/3574186.

• Nishiyama, I., Ikushima, T., and Ichikawa, S. (1966). Radiobiological studies in plants. XI. Further studies on somatic mutation induced by X-rays at the al locus of diploid oats. Rad. Bot. 6, 211–218. https://doi.org/10.1016/S0033-7560(66)80057-1.

• Park, I.S., Lee, G.J., Kim, D.S., Chung, S.J., Kim, J.B., Song, H.S., Goo, D.H., and Kang, S.Y. (2007). Mutation breeding of a spray chrysanthemum ‘Argus’ by gamma-ray irradiation and tissue culture. Flower Res. J. 15, 52–57.

• Solfanelli, C., Poggi, A., Loreti, E., Alpi, A., and Perata, P. (2006). Sucrose-specific induction of the anthocyanin biosynthetic pathway in Arabidopsis. Plant Physiol. 140, 637–646. https://doi.org/10.1104/pp.105.072579.

• Sripichtt, P., Nawata, E., and Shigenaga, S. (1988). The effects of exposure dose and dose rate of gamma radiation on in vitro shoot-forming capacity of cotyledon explants in red pepper. Jpn. J. Breed. 38, 27–34. https://doi.org/10.1270/jsbbs1951.38.27.

• Sung, S.Y., Kim, S.H., Velusamy, V., Lee, Y.M., Ha, B.K., Kim, J.B., and Kim, D.S. (2013). Comparative gene expression analysis in a highly anthocyanin pigmented mutant of colorless chrysanthemum. Mol. Biol. Rpt. 40, 5177–5189. https://doi.org/10.1007/s11033-013-2620-5.

• Tanaka, Y., Sasaki, N., and Ohmiya, A. (2008). Biosynthesis of plant pigments: anthocyanins, betalains and carotenoids. Plant J. 54, 733–749. https://doi.org/10.1111/j.1365-313X.2008.03447.x.

• Tanaka, A., Shikazono, N., and Hase, Y. (2010). Studies on biological effects of ion beans 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. https://doi.org/10.1269/jrr.09143.

• Yamaguchi, H., Shimizu, A., Degi, K., and Morishita, T. (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. https://doi.org/10.1270/jsbbs.58.331.

Received: 26 April 2016 | Accepted: 1 July 2016 | Published: 29 August 2016 | Available online: 29 August 2016