GENETIC DIVERSITY OF CHRYSANTHEMUM PLANTS DERIVED VIA SOMATIC EMBRYOGENESIS USING RAPD MARKERS
Justyna Lema-Rumińska
Laboratory of Biotechnology, Department of Ornamental Plants and Vegetable Crops, UTP University of Science and Technology, Bernardyńska 6, 85-029 Bydgoszcz, PolandAnna Mellem
Laboratory of Biotechnology, Department of Ornamental Plants and Vegetable Crops, UTP University of Science and Technology, Bernardyńska 6, 85-029 Bydgoszcz, PolandAbstract
The genetic diversity was investigated among two chrysanthemum (Chrysanthemum × grandiflorum Ramat./Kitam.) cultivars ‘Lady Salmon’ and ‘Lady Vitroflora’ and its 15 lines of plants derived from somatic embryos in using ten random amplified polymorphic DNA (RAPD) markers. All primers gave 108 bands with 1218 products from 148.65 to 4391.20 bp in size. The average number of bands per primer was 10.8. Most fragments (54; 50%) were polymorphic, 9 (0.8%) specific and others (45; 49.2%) were monomorphic. Cluster analysis grouped all the cultivars and their lines into two main clusters and two subclusters. Most genetic diversity was characteristic for LS2 lines of plants derived via somatic embryogenesis from cultivar ‘Lady Salmon’. All lines were different from each other and from the original cultivar propagated by meristematic explants. RAPD markers are a helpful tool to detect the genetic diversity of chrysanthemum plants derived via somatic embryogenesis (SE). Our result will provide useful information for production laboratory and for breeding programmes.
Keywords:
Chrysanthemum × grandiflorum, molecular markers, somatic embryos, genetic distanceReferences
Baliyan, D., Sirohi, A., Kumar, M., Kumar, V., Malik, S., Sharma, S., Sharma, S. (2014). Comparative genetic diversity analysis in chrysanthemum: A pilot study based on morpho-agronomic traits and ISSR markers. Sci. Hortic., 167, 164–168.
Broertjes, C., Roest, S., Bokelmann, G.S. (1976). Mutation breeding of Chrysanthemum morifolium Ram. using in vivo and in vitro adventitious bud techniques.
Euphytica, 25, 11–19.
Bush, S.R., Earle, E.D., Langhans, R.W. (1976). Plantlets from petal segments, petal epidermis and shoot tips of the periclinal chimera, Chrysanthemum morifolium
‘Indianapolis’. Am. J. Bot., 63, 729–737.
Chattarjee, J., Mandal, A.K.A., Ranade, S.A., Teixeira da Silva, J.A., Datta, S.K. (2006). Molecular systematic in Chrysanthemum × grandiflorum (Ramat.) Kitamura. Sci. Hortic., 110, 373–378.
Currais, L., Loureiro, J., Santos, C., Canhoto, J.M. (2013). Ploidy stability in embryogenic cultures and regenerated plantlets of tamarillo. Plant Cell Tiss. Organ Cult.,
114, 149–159.
Feng, S.G., Hea, R.F., Jianga, M.Y., Lua, J.J., Shenb, X.X., Liuc, J.J., Wangb, Z.A., Wanga, H.Z. (2016). Genetic diversity and relationships of medicinal Chrysanthemum
morifolium revealed by start codon targeted (SCoT) markers. Sci. Hortic., 201, 118–123.
Jerzy, M., Zalewska, M. (1996). Polish cultivars of Dendranthema grandiflora Tzvelev and Gerbera jamesonii Bolus bred in vitro by induced mutations. Mutat.
Breed. Newsl., 42, 19.
Jerzy, M., Zalewska, M. (1997). Flower colour recurrence in chrysanthemum and gerbera mutants propagated in vitro from meristems and leaf explants. Acta Hortic.,
447, 611–614.
Konieczny, R., Sliwinska, E., Pilarska, M., Tuleja, M. (2012). Morphohistological and flow cytometric analyses of somatic embryogenesis in Trifolium nigrescens Viv. Plant Cell Tiss. Organ Cult., 109, 131–141.
Lema-Rumińska, J., Zalewska, M. (2002). Ocena ploidalności mutantów chryzantemy wielkokwiatowej (Dendranthema grandiflora Tzvelev) uzyskanych w wyniku
mutagenezy indukowanej in vitro i in vivo promieniowaniem jonizującym. Acta Sci. Pol. Hortorum Cultus, 1(2), 43–48.
Lema-Rumińska, J., Zalewska, M., Sadoch, Z. (2004). Radiomutants of chrysanthemum (Dendranthema grandiflora Tzvelev) of Lady group: RAPD analysis of the genetic diversity. Plant Breed., 123(3), 290–293.
Lema-Rumińska, J., Niedojadło, J. (2014). Somatic embryogenesis in leaf explants of chrysanthemum radiomutants of the ‘Lady’ group. Propagation of Ornamental
Plants 14 (4), 177–183.
Lema-Rumińska, J., Śliwińska, E. (2015). Evaluation of the genetic stability of plants obtained via somatic embryogenesis in Chrysanthemum × grandiflorum
(Ramat./Kitam.). Acta Sci. Pol. Hortorum Cultus, 14(3), 131–139.
Li, T., Guo, J., Li, Y., Ning, H., Sun, X., Zheng, C. (2013). Genetic diversity assessment of chrysanthemum germplasm using conserved DNA-derived polymorphism
markers. Sci. Hort., 162, 271–277.
Lin, L. Z., Harnly, J. M. (2010). Identification of the phenolic components of chrysanthemum flower (Chrysanthemum morifolium Ramat). Food Chem., 120, 319–326.
Martin, C., Gonzales-Benito, M.E. (2005). Survival and genetic stability of Dendranthema grandiflora Tzvelev shoot apices after cryopreservation by vitrification and
encapsulation-dehydratation. Cryobiology, 51, 281–289.
Miler, N., Zalewska, M. (2014). Somaclonal variation of chrysanthemum propagated in vitro from different explants types. Acta Sci. Pol. Hortorum Cultus, 13(2),
69–82.
Miňano, H.S., González-Benito, M.E., Martín, C. (2009). Molecular characterization and analysis of somaclonal variation in chrysanthemum cultivars using RAPD
markers. Sci. Hort., 122, 238–243.
Mukherjee, A.K., Dey, A., Acharya, L., Palai, S.K., Panda, P.C. (2013). Studies on genetic diversity in elite varieties of Chrysanthemum using RAPD and ISSR markers.
Indian J. Biotechnol., 12, 161–169.
Myśków, B., Masojć, P., Banek-Tabor, A., Szołkowski, A. (2001). Genetic diversity of inbred rye lines evaluated by RAPD analysis. J. Applied Genet., 42(1), 1–14.
Naing, A.H., Kim, C.K., Yun, B.J., Jin, J.Y., Lim, K.B. (2013a). Primary and secondary somatic embryogenesis in Chrysanthemum cv. Euro. Plant Cell Tiss. Organ Cult., 112, 361–368.
Naing, A.H., Min, J.S., Park, K.I., Chung, M.Y., Lim, S.H., Lim, K.B., Kim, C.K. (2013b). Primary and secondary somatic embryogenesis in Chrysanthemum (Chrysanthemum
morifolium) cv. ‘Baeksun’ and assessment of ploidy stability of somatic embryogenesis process by flow cytometry. Acta Physiol. Plant., 35, 2965–2974.
Nei, M., Li, W.H. (1979). Mathematical model for studying genetical variation in terms of restriction endonucleases. Proc. Natl. Acad. Sci. U.S.A., 76, 5269–5273.
Pinto, D.L.P., Barros, B.A., Viccini, L.F., Salabert de Campos, J.M., da Silva, M.L., Otoni, W.C. (2010). Ploidy stability of somatic embryogenesis-derived Passiflora
cincinnata Mast. plants as assessed by flow cytometry. Plant Cell Tiss. Organ Cult., 103, 71–79.
Prado, M.J., Rodriguez, E., Rey, L., González, M.V., Santos, C., Rey, M. (2010). Detection of somaclonal variants in somatic embryogenesis regenerated plants of Vitis
vinifera by flow cytometry and microsatellite markers. Plant Cell Tiss. Organ Cult., 103, 49–59.
Roein, Z., Asil, M.H., Sabouri, A., Dadras, A.R. (2014). Genetic structure of Chrysanthemum genotypes from Iran assessed by AFLP markers and phenotypic traits.
Plant System. Evol., 300, 493–503.
Shibata, M., Kishimoto, S., Hirai, M., Aida, R., Ikeda, I. (1998). Analysis of the periclinal structure of chrysanthemum sports by random amplified polymorphic DNA. Acta Hortic., 454, 347–353.
Song, J.Y., Mattson, N.S., Jeong, B.R. (2011). Efficiency of shoot regeneration from leaf, stem, petiole and petal explants of six cultivars of Chrysanthemum morifolium.
Plant Cell Tiss. Organ Cult., 107, 295–304.
Stewart, R.N., Dermen, H. (1970). Somatic genetic analysis of the apical layers of chimeral sports in chrysanthemum by experimental production of adventitious shoots. Am. J. Bot., 57(9), 1061–1071.
Teixeira da Silva, J. A. (2004). Ornamental chrysanthemums: improvement by biotechnology. Plant Cell Tiss. Organ Cult., 79, 1–18.
Teixeira da Silva, J.A., Lema-Rumińska, J., Tymoszuk, A., Kulpa, D. (2015). Regeneration from chrysanthemum flowers: a review. Acta Physiol. Plant., 37, 36.
Van Harten, A. M. (1998). Mutation breeding. Theory and practical applications. Cambridge University Press, pp. 265–269.
Viehmannova, I., Bortlova, Z., Vitamvas, J., Cepkova, P.H., Eliasova, K., Svobodova, E., Travnickova, M. (2014). Assessment of somaclonal variation in somatic
embryo-derived plants of yacon [Smallanthus sonchifolius (Poepp. and Endl.) H. Robinson] using inter simple sequence repeat analysis and flow cytometry. Electronic
J. Biotech., 17, 102–106.
Wolff, K., Peters-Van Rijn, J. (1993). Rapid detection of genetic variability in chrysanthemum (Dendranthema grandiflora Tzvelev) using random primers. Heredity,
71, 335–341.
Wolff, K., Zietkiewicz, E., Hofstra, H. (1995). Identification of chrysanthemum cultivars and stability of DNA fingerprint patterns. Theor. App. Genet., 91, 439–447.
Wolff, K. (1996). RAPD analysis of sporting and chimerism in chrysanthemum. Euphytica, 89, 159–164.
Xu, P., Zhang, Z., Xia, X., Jia, J. (2012). Somatic embryogenesis and plant regeneration in chrysanthemum (Yuukou). Plant Cell Tiss. Organ Cult., 111, 393–397.
Zhang, Y., Dai, S., Hong, Y., Song, X. (2014). Application of Genomic SSR Locus Polymorphisms on the Identification and Classification of Chrysanthemum Cultivars
in China. PLoS ONE 9(8), e104856.
Zalewska, M., Lema-Rumińska, J., Miler, N., Gruszka, M., Dąbal, W. (2011). Induction of adventitious shoot regeneration in chrysanthemum as affected by the season.
In Vitro Cell. Dev. Biol. – Plant, 47, 375–378.
Zalewska, M., Lema-Rumińska, J., Miler, N. (2007). In vitro propagation using adventitious buds technique as a source of new variability in chrysanthemum. Sci.
Hortic., 113, 70–73.
Broertjes, C., Roest, S., Bokelmann, G.S. (1976). Mutation breeding of Chrysanthemum morifolium Ram. using in vivo and in vitro adventitious bud techniques.
Euphytica, 25, 11–19.
Bush, S.R., Earle, E.D., Langhans, R.W. (1976). Plantlets from petal segments, petal epidermis and shoot tips of the periclinal chimera, Chrysanthemum morifolium
‘Indianapolis’. Am. J. Bot., 63, 729–737.
Chattarjee, J., Mandal, A.K.A., Ranade, S.A., Teixeira da Silva, J.A., Datta, S.K. (2006). Molecular systematic in Chrysanthemum × grandiflorum (Ramat.) Kitamura. Sci. Hortic., 110, 373–378.
Currais, L., Loureiro, J., Santos, C., Canhoto, J.M. (2013). Ploidy stability in embryogenic cultures and regenerated plantlets of tamarillo. Plant Cell Tiss. Organ Cult.,
114, 149–159.
Feng, S.G., Hea, R.F., Jianga, M.Y., Lua, J.J., Shenb, X.X., Liuc, J.J., Wangb, Z.A., Wanga, H.Z. (2016). Genetic diversity and relationships of medicinal Chrysanthemum
morifolium revealed by start codon targeted (SCoT) markers. Sci. Hortic., 201, 118–123.
Jerzy, M., Zalewska, M. (1996). Polish cultivars of Dendranthema grandiflora Tzvelev and Gerbera jamesonii Bolus bred in vitro by induced mutations. Mutat.
Breed. Newsl., 42, 19.
Jerzy, M., Zalewska, M. (1997). Flower colour recurrence in chrysanthemum and gerbera mutants propagated in vitro from meristems and leaf explants. Acta Hortic.,
447, 611–614.
Konieczny, R., Sliwinska, E., Pilarska, M., Tuleja, M. (2012). Morphohistological and flow cytometric analyses of somatic embryogenesis in Trifolium nigrescens Viv. Plant Cell Tiss. Organ Cult., 109, 131–141.
Lema-Rumińska, J., Zalewska, M. (2002). Ocena ploidalności mutantów chryzantemy wielkokwiatowej (Dendranthema grandiflora Tzvelev) uzyskanych w wyniku
mutagenezy indukowanej in vitro i in vivo promieniowaniem jonizującym. Acta Sci. Pol. Hortorum Cultus, 1(2), 43–48.
Lema-Rumińska, J., Zalewska, M., Sadoch, Z. (2004). Radiomutants of chrysanthemum (Dendranthema grandiflora Tzvelev) of Lady group: RAPD analysis of the genetic diversity. Plant Breed., 123(3), 290–293.
Lema-Rumińska, J., Niedojadło, J. (2014). Somatic embryogenesis in leaf explants of chrysanthemum radiomutants of the ‘Lady’ group. Propagation of Ornamental
Plants 14 (4), 177–183.
Lema-Rumińska, J., Śliwińska, E. (2015). Evaluation of the genetic stability of plants obtained via somatic embryogenesis in Chrysanthemum × grandiflorum
(Ramat./Kitam.). Acta Sci. Pol. Hortorum Cultus, 14(3), 131–139.
Li, T., Guo, J., Li, Y., Ning, H., Sun, X., Zheng, C. (2013). Genetic diversity assessment of chrysanthemum germplasm using conserved DNA-derived polymorphism
markers. Sci. Hort., 162, 271–277.
Lin, L. Z., Harnly, J. M. (2010). Identification of the phenolic components of chrysanthemum flower (Chrysanthemum morifolium Ramat). Food Chem., 120, 319–326.
Martin, C., Gonzales-Benito, M.E. (2005). Survival and genetic stability of Dendranthema grandiflora Tzvelev shoot apices after cryopreservation by vitrification and
encapsulation-dehydratation. Cryobiology, 51, 281–289.
Miler, N., Zalewska, M. (2014). Somaclonal variation of chrysanthemum propagated in vitro from different explants types. Acta Sci. Pol. Hortorum Cultus, 13(2),
69–82.
Miňano, H.S., González-Benito, M.E., Martín, C. (2009). Molecular characterization and analysis of somaclonal variation in chrysanthemum cultivars using RAPD
markers. Sci. Hort., 122, 238–243.
Mukherjee, A.K., Dey, A., Acharya, L., Palai, S.K., Panda, P.C. (2013). Studies on genetic diversity in elite varieties of Chrysanthemum using RAPD and ISSR markers.
Indian J. Biotechnol., 12, 161–169.
Myśków, B., Masojć, P., Banek-Tabor, A., Szołkowski, A. (2001). Genetic diversity of inbred rye lines evaluated by RAPD analysis. J. Applied Genet., 42(1), 1–14.
Naing, A.H., Kim, C.K., Yun, B.J., Jin, J.Y., Lim, K.B. (2013a). Primary and secondary somatic embryogenesis in Chrysanthemum cv. Euro. Plant Cell Tiss. Organ Cult., 112, 361–368.
Naing, A.H., Min, J.S., Park, K.I., Chung, M.Y., Lim, S.H., Lim, K.B., Kim, C.K. (2013b). Primary and secondary somatic embryogenesis in Chrysanthemum (Chrysanthemum
morifolium) cv. ‘Baeksun’ and assessment of ploidy stability of somatic embryogenesis process by flow cytometry. Acta Physiol. Plant., 35, 2965–2974.
Nei, M., Li, W.H. (1979). Mathematical model for studying genetical variation in terms of restriction endonucleases. Proc. Natl. Acad. Sci. U.S.A., 76, 5269–5273.
Pinto, D.L.P., Barros, B.A., Viccini, L.F., Salabert de Campos, J.M., da Silva, M.L., Otoni, W.C. (2010). Ploidy stability of somatic embryogenesis-derived Passiflora
cincinnata Mast. plants as assessed by flow cytometry. Plant Cell Tiss. Organ Cult., 103, 71–79.
Prado, M.J., Rodriguez, E., Rey, L., González, M.V., Santos, C., Rey, M. (2010). Detection of somaclonal variants in somatic embryogenesis regenerated plants of Vitis
vinifera by flow cytometry and microsatellite markers. Plant Cell Tiss. Organ Cult., 103, 49–59.
Roein, Z., Asil, M.H., Sabouri, A., Dadras, A.R. (2014). Genetic structure of Chrysanthemum genotypes from Iran assessed by AFLP markers and phenotypic traits.
Plant System. Evol., 300, 493–503.
Shibata, M., Kishimoto, S., Hirai, M., Aida, R., Ikeda, I. (1998). Analysis of the periclinal structure of chrysanthemum sports by random amplified polymorphic DNA. Acta Hortic., 454, 347–353.
Song, J.Y., Mattson, N.S., Jeong, B.R. (2011). Efficiency of shoot regeneration from leaf, stem, petiole and petal explants of six cultivars of Chrysanthemum morifolium.
Plant Cell Tiss. Organ Cult., 107, 295–304.
Stewart, R.N., Dermen, H. (1970). Somatic genetic analysis of the apical layers of chimeral sports in chrysanthemum by experimental production of adventitious shoots. Am. J. Bot., 57(9), 1061–1071.
Teixeira da Silva, J. A. (2004). Ornamental chrysanthemums: improvement by biotechnology. Plant Cell Tiss. Organ Cult., 79, 1–18.
Teixeira da Silva, J.A., Lema-Rumińska, J., Tymoszuk, A., Kulpa, D. (2015). Regeneration from chrysanthemum flowers: a review. Acta Physiol. Plant., 37, 36.
Van Harten, A. M. (1998). Mutation breeding. Theory and practical applications. Cambridge University Press, pp. 265–269.
Viehmannova, I., Bortlova, Z., Vitamvas, J., Cepkova, P.H., Eliasova, K., Svobodova, E., Travnickova, M. (2014). Assessment of somaclonal variation in somatic
embryo-derived plants of yacon [Smallanthus sonchifolius (Poepp. and Endl.) H. Robinson] using inter simple sequence repeat analysis and flow cytometry. Electronic
J. Biotech., 17, 102–106.
Wolff, K., Peters-Van Rijn, J. (1993). Rapid detection of genetic variability in chrysanthemum (Dendranthema grandiflora Tzvelev) using random primers. Heredity,
71, 335–341.
Wolff, K., Zietkiewicz, E., Hofstra, H. (1995). Identification of chrysanthemum cultivars and stability of DNA fingerprint patterns. Theor. App. Genet., 91, 439–447.
Wolff, K. (1996). RAPD analysis of sporting and chimerism in chrysanthemum. Euphytica, 89, 159–164.
Xu, P., Zhang, Z., Xia, X., Jia, J. (2012). Somatic embryogenesis and plant regeneration in chrysanthemum (Yuukou). Plant Cell Tiss. Organ Cult., 111, 393–397.
Zhang, Y., Dai, S., Hong, Y., Song, X. (2014). Application of Genomic SSR Locus Polymorphisms on the Identification and Classification of Chrysanthemum Cultivars
in China. PLoS ONE 9(8), e104856.
Zalewska, M., Lema-Rumińska, J., Miler, N., Gruszka, M., Dąbal, W. (2011). Induction of adventitious shoot regeneration in chrysanthemum as affected by the season.
In Vitro Cell. Dev. Biol. – Plant, 47, 375–378.
Zalewska, M., Lema-Rumińska, J., Miler, N. (2007). In vitro propagation using adventitious buds technique as a source of new variability in chrysanthemum. Sci.
Hortic., 113, 70–73.
Justyna Lema-Rumińska
Laboratory of Biotechnology, Department of Ornamental Plants and Vegetable Crops, UTP University of Science and Technology, Bernardyńska 6, 85-029 Bydgoszcz, Poland
Laboratory of Biotechnology, Department of Ornamental Plants and Vegetable Crops, UTP University of Science and Technology, Bernardyńska 6, 85-029 Bydgoszcz, Poland
Anna Mellem
Laboratory of Biotechnology, Department of Ornamental Plants and Vegetable Crops, UTP University of Science and Technology, Bernardyńska 6, 85-029 Bydgoszcz, Poland
Laboratory of Biotechnology, Department of Ornamental Plants and Vegetable Crops, UTP University of Science and Technology, Bernardyńska 6, 85-029 Bydgoszcz, Poland
License
Articles are made available under the conditions CC BY 4.0 (until 2020 under the conditions CC BY-NC-ND 4.0).
Submission of the paper implies that it has not been published previously, that it is not under consideration for publication elsewhere.
The author signs a statement of the originality of the work, the contribution of individuals, and source of funding.
