Article | 12. 2015 Vol. 33, Issue. 6
The Selection of Domestically Bred Cultivars for Spray-type Chrysanthemum Transformation



Molecular breeding Division, National Academy of Agricultural Science, Rural Development Administration1
Department of Seed Service, The Foundation of Agri. Tech. Commercialization and Transfer2




2015.12. 941:954


PDF XML




To select suitable spray chrysanthemum cultivars for Agrobacterium-mediated transformation, thirty-nine (39) spray cultivars bred in the National Institutes of Korea and a standard cultivar Jinba from Japan were collected and tested for regeneration rate and Agrobacterium infection assays. MS medium with 0.5 mg・L-1 IAA and 1.0 mg・L-1 BAP was used for shoot regeneration from leaf disks and internodes. The shoot regeneration rate in leaf disks was the highest in cultivar BRM, followed by cultivars VS, WW and YTM. The cultivar JB (Jinba) used as a transformation material in previous reports ranked similarly to cultivars PK and SPP. In shoot regeneration from internodes, the shoot regeneration rate was the highest for cultivar PA, followed by cultivar WW. The infection rate of leaves and internodes of 40 chrysanthemum cultivars with agrobacterium was investigated. Cultivars WPP, YNW, VS, PP, WW, FA, PA and YMN showed the highest infection levels in leaves, whereas cultivars WPP, PA, PK and YNW had the highest infection levels in internodes. Considering all of these results, cultivars VS and WW were the most appropriate for gene transformation of chrysanthemum using leaves, while cultivar PA was for internodes.



1. Aida, R., Y. Tabei, M. Hirai, and M. Shibata. 1992. Agrobacterium- mediated transformation of chrysan-themum. Breed. Sci. 42: 270-271.   

2. An, J., A. Song, Z. Guan, J. Jiang, F. Chen, W. Lou, W. Fang, Z. Liu, and S. Chen. 2014. The over-expression of Chrysanthemum crassum CcSOS1 improves the salinity tolerance of chrysanthemum. Mol. Biol. Rep. 41:4155-4162.  

3. Chen, L. 2005. Research and analysis of the international market of chrysanthemum. Greenhouse Hortic. 8:20-22.  

4. De Jong, J., W. Rademaker, and K. Ohishi. 1995. Agrobacterium- mediated transformation of chrysan-themum. Plant Tissue Cult. Biotechnol. 1:38-42.  

5. Duncan, D.B. 1955. Multiple range and multiple F test. Biometrics 11:1-42.  

6. Fukai, S., J. De Jong, and W. Rademaker. 1995. Efficient genetic transformation of chrysanthemum (Dendranthema grandiforum (Ramat.) Kitamura) using stem segment. Breed. Sci. 45:179-184.  

7. Han, B.H., E.J. Suh, S.Y. Lee, H.K. Shin, and Y.P. Lim. 2007. Selection of non-branching lines induced by introducing Ls-like cDNA into chrysanthemum (Dendranthema × grandiflorum (Ramat.) Kitamura) ‘Shuho-no-chikara’. Sci. Hortic. 115:70-75.  

8. Han, B.H., S.Y. Lee, and B.M. Park. 2009. Comparison of chrysanthemum cultivars based on direct shoot regeneration rates in tissue culture. J. Plant Biotechnol. 36:275-280.  

9. Hwang, J.C., Y.D. Chin, Y.M. Chung, S.K. Kim, C.W. Ro, and B.R. Jeong. 2013. A new spray chrysanthemum cultivar, ‘Blue Hope’ with anemone type and white petals for cut flower. Korean J. Hortic. Sci. Technol. 31:123-127.  

10. Jefferson, R.A. 1987. Plant reporter genes: The gus gene fusion system. Genetic Engineering 10:247-263.  

11. Kim, Y.H., H.M. Park, J.Y. Jung, T.M. Kwon, S.J. Jeung, Y.B. Yi, G.T. Kim, and J.S. Nam. 2010. Development of Agrobacterium- mediated transformation method for domestically bred chry-santhemum cultivar ‘Moulinrouge’ and genetic change of leaf morphology using AtSICKLE gene. Korean J. Hortic. Sci. Technol. 28:449-455.  

12. Kosugi, S., Y. Ohashi, K. Nakajima, and A. Yuji. 1990. An improved assay for β-glucuronidase in trans-formed cells: Methanol almost completely suppresses a putative endogenous β-glucuronidase. Plant Sci. 70:133-140.  

13. Ledger, S.E., S.C. Deroles, and N.K. Given. 1991. Regeneration and Agrobacterium-mediated transformation of chrysanthemum. Plant Cell Rep. 10:195-199.  

14. Lee, S.Y., B.H. Han, E.J. Hur, H.K. Shin, S.T. Kim, E.K. Lee, W.H. Kim, O.H. Kwon, and I.H. Lee. 2012. FT-transgenic spray-type chrysanthemum (Dendranthema grandiflorum Kitamura) showing early flowering. J. Plant Biotechnol. 39:140-145.  

15. Lee, S.Y., J.H. Kim, K.S. Cheon, E.K. Lee, W.H. Kim, O.H. Kwon, and H.J. Lee. 2013. Phenotypic and molecular characteristics of second clone (TV) plants of the LeLs-antisense gene- transgenic chrysanthemum line exhibiting non-branching. J. Plant Biotechnol. 40:192-197.  

16. Li, Y.F., W.M. Fang, F.D. Chen, S.M. Chen, and C.L. Shi. 2009. Effect of different planting date on phenophase and quality of spray cut chrysanthemum produced in summer, J. Yangzhou University, Agricultural and Life Sciences ed. 30:80-83.  

17. Miller, M. 1975. Leaf, stem, crown, and root galls induced in chrysanthemum by Agrobacterium tumefaciens. Phytopathology 65:805-811.  

18. Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:473-497.  

19. Shinoyama, H. and A. Mochizuki. 2006. Insect resistant transgenic- chrysanthemum [Dendranthema × grandiflorum (Ramat.) Kitamura]. Acta Hortic. 714:177-184.  

20. Shinoyama, H., R. Aida, R. Ichikawa, H. Ichikawa, Y. Nomura, and Y. Mochizuki. 2012. Genetic engineering of chrysanthemum (Chrysanthemum morifolium): Current progress and perspectives. Plant Biotechnol. 29:323-337.  

21. Song, A., J. An, Z. Guan, J. Jiang, F. Chen, W. Lou W. Fang, Z. Liu, and C. Sumei. 2014. The constitutive expression of a two transgene construct enhances the abiotic stress tolerance of chrysanthemum. Plant Physiol. Biochem. 80:114-120.  

22. Takatsu, Y., H. Tomotsune, M. Kasumi, and F. Sakuma. 1998. Differences in adventitious shoot regeneration capacity among Japanese chrysanthemum (Dendranthema grandiflora (Ramat.) Kitamura) cultivars and the improved protocol for Agrobacterium- mediated genetic transformation. J. Jpn. Soc. Hortic. Sci. 67:958-964.  

23. Teixeira da Silva, J.A. 2005. Effective and comprehensive chry-santhemum (Dendranthema × grandiflora) regeneration and transformation protocols. Biotechnology 4:94-107.  

24. Teixeira da Silva, J. A., H. Shinoyama, R. Aida, Y. Matsushita, S.K. Raj, and F. Chen. 2013. Chrysanthemum biotechnology: Quo vadis? Crit. Rev. Plant. Sci. 32:21-52.  

25. Teixeira da Silva, J.A. and S. Fukai. 2004. Effect of amino-glycoside antibiotics on in vitro morphogenesis from cultured cells of chrysanthemum and tobacco. J. Plant Biotechnol. 6:25-37.  

26. Urban, L.A., J.M. Sherman, J.W. Moyer, and M.E. Daub. 1994. High frequency shoot regeneration and Agrobacterium-mediated transformation of chrysanthemum (Dendranthema grandiflora). Plant Sci. 98:69-79.