Abstract
Spring orchid, Cymbidium goeringii, is one of the most important and popular species in the orchid family in Northeast Asia. The purpose of this study was to determine how closely the simple sequence repeat (SSR) genotypes match between individual plants assumed to be the same cultivar in C. goeringii. For this study, we collected 1566 samples from 17 C. goeringii cultivars of the flower-variant types and determined the frequencies and distribution of the combined genotypes of 13 SSR loci by the multiplex PCR method. The phylogenetic distance matrix and the principal coordinate analysis revealed that the correlation between horticultural phenotypes and genetic distance was loosely matched, but two cultivars of Japanese origin were clearly separated from the Korean cultivars. The results showed that the ratio of the most frequent combined genotypes in 10 cultivar groups is approximately 60%, which suggests that almost 40% of C. goeringii sold may not be genuine. This is the first forensic attempt to authenticate whether each plant is genetically genuine for the corresponding cultivars in spring orchid. We suggest preparation of a guideline for DNA deposit and profiling of the SSR genotypes for the newly registered C. goeringii cultivars and believe that this study will be helpful for establishing a molecular genetic method for forensic authentication of these cultivars.
Similar content being viewed by others
Data availability
The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
References
Bär W, Brinkmann B, Budowle B, Carracedo A, Gill P, Lincoln P, Mayr W, Olaisen B (1997) DNA recommendations-further report of the DNA commission of the ISFH regarding the use of short tandem repeat systems. Int J Legal Med 110:175–176
Choi SH, Kim MJ, Lee JS, Ryu KH (2006) Genetic diversity and phylogenetic relationships among and within species of oriental Cymbidiums based on RAPD analysis. Sci Hortic 108:79–85
Chung MY, Chung MG (1999) Allozyme diversity and population structure in Korean populations of Cymbidium goeringii (Orchidaceae). J Plant Res 112:139–144
Dressler RL (1993) Phylogeny and classification of the orchid family. Cambridge University Press, Cambridge
DuPuy D, Cribb PJ (1988) The genus Cymbidium. Christopher Helm, London
Garay LA, Sweet HR (1974) Natural and artificial hybrid generic names of orchid. In: Withner CL (ed) The orchids, scientific studies. Wiley, New York, pp 485–561
Huang JL, Zeng CX, Li HT, Yang JB (2011) Isolation and characterization of 15 microsatellite markers from the spring orchid (Cymbidium goeringii) (Orchidaceae). Am J Bot 98:e76–e77
Hyun YS, Kim J, Chung KW (2012) Development of polymorphic microsatellite markers for Cymbidium goeringii (Orchidaeceae). Am J Bot 99:e193–e198
Jian L, Zhang Y-Z, Yu D-F, Zhu L-Q (2010) Molecular characterization of Cymbidium kanran cultivars based on extended random amplified polymorphic DNA (ERAPD) markers. Afr J Biotechnol 9:5084–5089
Lee JS, Kim BM, Yu SO, Ryu KH (2004) Analysis of phenotypic and genetic polymorphism of self-pollinated seedlings of Korean native Cymbidium goeringii. Korean J Hortic Sci Technol 22:486–490
Lee DG, Koh JC, Chung KW (2012) Determination and application of combined genotype of simple sequence repeats (SSR) DNA marker for cultivars of Cymbidium goeringii. Korean J Hortic Sci Technol 30:278–285
Li X, Jin F, Jin L, Jackson A, Huang C, Li K, Shu X (2014) Development of Cymbidium ensifolium genic-SSR markers and their utility in genetic diversity and population structure analysis in cymbidiums. BMC Genet 5:124
Liu X, Huang Y, Li F, Xu C, Chen K (2014) Genetic diversity of 129 spring orchid (Cymbidium goeringii) cultivars and its relationship to horticultural types as assessed by EST-SSR markers. Sci Hortic 174:178–184
Lu J, Hu X, Liu J, Wang H (2011) Genetic diversity and population structure of 151 Cymbidium sinense cultivars. J Hortic For 3:104–114
Moe KT, Zhao W, Song H-S, Kim Y-H, Chung J-W, Cho Y-I, Park PH, Park H-S, Chae S-C, Park Y-J (2010) Development of SSR markers to study diversity in the genus Cymbidium. Biochem Syst Ecol 38:585–594
Obara-Okeyo P, Kako S (1998) Genetic diversity and identification of Cymbidium cultivars as measured by random amplified polymorphic DNA (RAPD) markers. Euphytica 99:95–101
Park HY, Kang KW, Kim DH, Sivanesan I (2018a) In vitro propagation of Cymbidium goeringii Reichenbach fil. through direct adventitious shoot regeneration. Physiol Mol Biol Plants 24:307–313
Park SY, Kim DY, Park YH, Lee JH, Kwon SW (2018b) A fragrant Cymbidium goeringii variety 'Arihyang' with a single jade flower in a peduncle and plain leaf. Korean J Breed Sci 50:177–180
Peakall R, Smouse PE (2012) GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research-an update. Bioinformatics 28:2537–2539
Ryu J-H, Lee H-Y, Bae C-H (2011) Variation analysis of long-term in vitro cultured Cymbidium goeringii lindley and Cymbidium kanran Makino. Korean J Plant Res 24:139–149
Suetsugu K (2015) Autonomous self-pollination and insect visitors in partially and fully mycoheterotrophic species of Cymbidium (Orchidaceae). J Plant Res 128:115–125
Tang M, Zeng CX, Bi YF, Yang JB (2012) Microsatellite markers for the Chinese endangered and endemic orchid Cymbidium tortisepalum (Orchidaceae). Am J Bot 99:e11–e13
Wang HZ, Wang YD, Zhou XY, Ying QC, Zheng KL (2004) Analysis of genetic diversity of 14 species of Cymbidium based on RAPDs and AFLPs. Shi Yan Sheng Wu Xue Bao 37:482–486
Wang HZ, Wu ZX, Lu JJ, Shi NN, Zhao Y, Zhang ZT, Liu JJ (2009) Molecular diversity and relationships among Cymbidium goeringii cultivars based on inter-simple sequence repeat (ISSR) markers. Genetica 136:391–399
Wu ZX, Shi NN, Zhao Y, Wang HZ (2008) Study on the relationships of Cymbidium by RMAPD technique. Fen Zi Xi Bao Sheng Wu Xue Bao 41:145–149
Xiaohong Y, Li G, Bo Y (2007) Genetic diversity of wild Cymbidium goeringii (Orchidaceae) populations from Hubei based on inter-simple sequence repeats analysis. Front Biol China 2:419–424
Acknowledgements
We would like to thank the cultivators who provided the spring orchid samples, the Korea Registration Association for providing the registration information, and the World of Orchid for providing orchid images. This work was supported by a grant from Kongju National University (2018).
Author information
Authors and Affiliations
Contributions
HJL, HRP and AJL performed molecular genetic works. DGL collected samples. DEN and YD performed the statistical analysis and preparation of manuscript. KWC designed this study and preparation of manuscript.
Corresponding author
Ethics declarations
Conflict of interest
All the authors declare that there are no conflicts of interest to disclose.
Additional information
Communicated by Inhwa Yeam.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Lee, H.J., Park, H.R., Lee, A.J. et al. Genetic authentication of cultivars with flower-variant types using SSR markers in spring orchid, Cymbidium goeringii. Hortic. Environ. Biotechnol. 61, 577–590 (2020). https://doi.org/10.1007/s13580-020-00243-9
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13580-020-00243-9