Journal of Ginseng Research

The Korean Society of Ginseng (고려인삼학회)
권호 표지
DOI QR코드

DOI QR Code

High frequency somatic embryogenesis and plant regeneration of interspecific ginseng hybrid between Panax ginseng and Panax quinquefolius

  • Kim, Jong Youn (Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University) ;
  • Adhikari, Prakash Babu (Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University) ;
  • Ahn, Chang Ho (Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University) ;
  • Kim, Dong Hwi (Department of Herbal Crop Research, National Institute of Horticulture and Herbal Science, Rural Development Administration) ;
  • Kim, Young Chang (Department of Herbal Crop Research, National Institute of Horticulture and Herbal Science, Rural Development Administration) ;
  • Han, Jung Yeon (Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University) ;
  • Kondeti, Subramanyam (Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University) ;
  • Choi, Yong Eui (Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University)
  • Received : 2016.10.18
  • Accepted : 2017.08.01
  • Published : 2019.01.15
Download PDF
⟨ Previous Next ⟩

Abstract

Background: Interspecific ginseng hybrid, Panax ginseng ${\times}$ Panax quenquifolius (Pgq) has vigorous growth and produces larger roots than its parents. However, F1 progenies are complete male sterile. Plant tissue culture technology can circumvent the issue and propagate the hybrid. Methods: Murashige and Skoog (MS) medium with different concentrations (0, 2, 4, and 6 mg/L) of 2,4-dichlorophenoxyacetic acid (2,4-D) was used for callus induction and somatic embryogenesis (SE). The embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 Schenk and Hildebrandt (SH) medium. The developed taproots with dormant buds were treated with $GA_3$ to break the bud dormancy, and transferred to soil. Hybrid Pgq plants were verified by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses and by LC-IT-TOF-MS. Results: We conducted a comparative study of somatic embryogenesis (SE) in Pgq and its parents, and attempted to establish the soil transfer of in vitro propagated Pgq tap roots. The Pgq explants showed higher rate of embryogenesis (~56% at 2 mg/L 2,4-D concentration) as well as higher number of embryos per explants (~7 at the same 2,4-D concentration) compared to its either parents. The germinated embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 SH medium to support the continued growth and kept until nutrient depletion induced senescence (NuDIS) of leaf defoliation occurred (4 months). By that time, thickened tap roots with well-developed lateral roots and dormant buds were obtained. All Pgq tap roots pretreated with 20 mg/L $GA_3$ for at least a week produced new shoots after soil transfer. We selected the discriminatory RAPD and ISSR markers to find the interspecific ginseng hybrid among its parents. The $F_1$ hybrid (Pgq) contained species specific 2 ginsenosides (ginsenoside Rf in P. ginseng and pseudoginsenosides $F_{11}$ in P. quinquefolius), and higher amount of other ginsenosides than its parents. Conclusion: Micropropagation of interspecific hybrid ginseng can give an opportunity for continuous production of plants.

Keywords

References

  1. Sharma SK, Pandit MK. A new species of Panax L. (Araliaceae) from Sikkim Himalaya. India Syst Bot 2009;34:434-8. https://doi.org/10.1600/036364409788606235
  2. Chan TWD, But PPH, Cheng SW, Kwok IMY, Lau FW, Xu HX. Differentiation and authentication of Panax ginseng, Panax quinquefolius, and ginseng products by using HPLC/MS. Anal Chem 2000;72:1281-7. https://doi.org/10.1021/ac990819z
  3. Choi KT. Botanical characteristics, pharmacological effects and medicinal components of Korean Panax ginseng C A Meyer. Acta Pharmacol Sin 2008;29:1109-18. https://doi.org/10.1111/j.1745-7254.2008.00869.x
  4. Kim JH, Jung JY, Choi HI, Kim NH, Park JY, Lee Y, Yang TJ. Diversity and evolution of major Panax species revealed by scanning the entire chloroplast intergenic spacer sequences. Genet Resour Crop Evol 2013;60:413-25. https://doi.org/10.1007/s10722-012-9844-4
  5. Christensen LP. Ginsenosides chemistry, biosynthesis, analysis, and potential health effects. Adv Food Nutr Res 2009;55:1-99. https://doi.org/10.1016/S1043-4526(08)00401-4
  6. Kim Y, Silva J, Zhang D, Shi J, Joo S, Jang M, Kwon W, Yang D. Development of interspecies hybrids to increase ginseng biomass and ginsenoside yield. Plant Cell Rep 2016;35:779-90. https://doi.org/10.1007/s00299-015-1920-8
  7. Jia L, Zhao Y. Current evaluation of the millennium phytomedicine - ginseng (I): etymology, pharmacognosy, phytochemistry, market and regulations. Curr Med Chem 2009;16:2475-84. https://doi.org/10.2174/092986709788682146
  8. Li L, Luo GA, Liang QL, Hu P, Wang YM. Rapid qualitative and quantitative analyses of Asian ginseng in adulterated American ginseng preparations by UPLC/Q-TOF-MS. J Pharm Biomed Anal 2010;52:66-72. https://doi.org/10.1016/j.jpba.2009.12.017
  9. Kim DH. Chemical diversity of Panax ginseng, Panax quinquifolium, and Panax notoginseng. J Ginseng Res 2012;36:1-15. https://doi.org/10.5142/jgr.2012.36.1.1
  10. Yue PYK, Mak NK, Cheng YK, Leung KW, Ng TB, Fan DTP, Yeung HW, Wong RNS. Pharmacogenomics and the Yin/Yang actions of ginseng: antitumor, angiomodulating and steroid-like activities of ginsenosides. Chin Med 2007;2:6-27. https://doi.org/10.1186/1749-8546-2-6
  11. Qi L-W, Wang C-Z, Yuan C-S. American ginseng: Potential structureefunction relationship in cancer chemoprevention. Biochem Pharmacol 2010;80:947-54. https://doi.org/10.1016/j.bcp.2010.06.023
  12. Qin J, Leung FC, Fung Y, Zhu D, Lin B. Rapid authentication of ginseng species using microchip electrophoresis with laser-induced fluorescence detection. Anal Bioanal Chem 2005;381:812-9. https://doi.org/10.1007/s00216-004-2889-2
  13. Ahn IO, Lee SS, Lee JH, Lee BS, In JG, Yang DC. Characteristics of plantlets redifferentiated from F1 hybrid between Panax ginseng and Panax quinquefolius. Korean J Plant Biotechnol 2006;33:45-8. https://doi.org/10.5010/JPB.2006.33.1.045
  14. Chung Y. Germination of hybrid ginseng seeds, and activities of lipoxygenase (LOX) in Panax ginseng species. J Ginseng Res 2004;28:191-5. https://doi.org/10.5142/JGR.2004.28.4.191
  15. Kitanaka S, Kunishi M, Taira T, Takahashi S, Yazawa S, Miyazawa Y, Nakajima Y, Okuno K, Fukuoka S, Takido M. Ginsenoside production, tissue culture and introduction of regeneration of interspecific hybrid ginseng Panax ginseng  P. quinquefolium. Natural Medicines 1997:1-3.
  16. Washida D, Shimomura K, Kitanaka S. Polyacetylenes in hairy roots of a Panax hybrid. Planta Med 2003;69:1163-5. https://doi.org/10.1055/s-2003-818012
  17. Duarte-Ake F, De-la-Pena C. Epigenetic advances in somatic embryogenesis in sequenced genome crops. In: Loyola-Vargas MV, Ochoa-Alejo N, editors. Somatic embryogenesis: fundamental aspects and applications. Cham: Springer International Publishing; 2016. p. 81-102.
  18. Chang WC, Hsing YI. Plant regeneration through somatic embryogenesis in root-derived callus of ginseng (Panax ginseng C. A. Meyer). Theor Appl Genet 1980;57:133-5. https://doi.org/10.1007/BF00253888
  19. Choi YE, Kim HS, Soh WY, Yang DC. Developmental and structural aspects of somatic embryos formed on medium containing 2,3,5-triiodobenzoic acid. Plant Cell Rep 1997;16:738-44. https://doi.org/10.1007/s002990050312
  20. Choi YE, Yang DC, Park JC, Soh WY, Choi KT. Regenerative ability of somatic single and multiple embryos from cotyledons of Korean ginseng on hormonefree medium. Plant Cell Rep 1998;17:544-51. https://doi.org/10.1007/s002990050439
  21. Choi YE, Yang DC, Yoon ES, Choi KT. High-efficiency plant production via direct somatic single embryogenesis from preplasmolysed cotyledons of Panax ginseng and possible dormancy of somatic embryos. Plant Cell Rep 1999;18:493-9. https://doi.org/10.1007/s002990050610
  22. Shoyama Y, Kamura K, Nishioka I. Somatic embryogenesis and clonal multiplication of Panax ginseng. Planta Med 1988;54:155-6. https://doi.org/10.1055/s-2006-962376
  23. Tirajoh A, Kyung TS, Punja ZK. Somatic embryogenesis and plantlet regeneration in American ginseng (Panax quinquefolium L.). Vitro Cell Dev Biol Plant 1998;34:203-11. https://doi.org/10.1007/BF02822709
  24. Uchendu EE, Paliyath G, Brown DCW, Saxena PK. In vitro propagation of North American ginseng (Panax quinquefolius L.). Vitro Cell Dev Biol Plant 2011;47:710-8. https://doi.org/10.1007/s11627-011-9379-y
  25. Zhou S, Brown DCW. High efficiency plant production of North American ginseng via somatic embryogenesis from cotyledon explants. Plant Cell Rep 2006;25:166-73. https://doi.org/10.1007/s00299-005-0043-z
  26. Murashige T, Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 1962;15:473-97. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  27. Kim JY, Kim DH, Kim YC, Kim KH, Han JY, Choi YE. In vitro grown thickened taproots, a new type of soil transplanting source in Panax ginseng. J Ginseng Res 2016;40:409-14. https://doi.org/10.1016/j.jgr.2016.05.003
  28. Schenk RU, Hildebrandt AC. Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures. Can J Bot 1972;50:199-204. https://doi.org/10.1139/b72-026
  29. Ahn CH, Kim YS, Lim S, Yi JS, Choi YE. Random amplified polymorphic DNA (RAPD) analysis and RAPD-derived sequence characterized amplified regions (SCAR) marker development to identify Chinese and Korean ginseng. J Med Plants Res 2011;5:4487-92.
  30. Choi YE, Yang DC, Yoon ES, Choi KT. Plant regeneration via adventitious bud formation from cotyledon explants of Panax ginseng C. A. Meyer. Plant Cell Rep 1998;17:731-6. https://doi.org/10.1007/s002990050474
  31. Zhou S, Brown DCW. A highly efficient protocol for micropropagation of North American ginseng. In: Xu Z, et al., editors. Biotechnology and sustainable agriculture 2006 and beyond: Proceedings of the 11th IAPTC&B Congress, August 31-18, 2006 Beijing, China. Dordrecht. Netherlands: Springer; 2007. p. 425-8.
  32. Choi YE. Ginseng. In: Pua EC, Davey RM, editors. Transgenic crops VI. Berlin, Heidelberg: Springer; 2007. p. 149-68.
  33. Furuya T, Yoshikawa T, Ishii T, Kajii K. Effects of auxins on growth and saponin production in callus cultures of Panax ginseng. Planta Med 1983;47:183-7. https://doi.org/10.1055/s-2007-969981
  34. Washida D, Shimomura K, Nakajima Y, Takido M, Kitanaka S. Ginsenosides in hairy roots of a panax hybrid. Phytochemistry 1998;49:2331-5. https://doi.org/10.1016/S0031-9422(98)00308-2
  35. Lim HT, Lee HS, Eriksson T. Regeneration of Panax ginseng C.A. Meyer by organogenesis and nuclear DNA analysis of regenerants. Plant Cell Tiss Org Cult 1997;49:179-87. https://doi.org/10.1023/A:1005820210895
  36. Raemakers CJJM, Jacobsen E, Visser RGF. Secondary somatic embryogenesis and applications in plant breeding. Euphytica 1995;81:93-107. https://doi.org/10.1007/BF00022463
  37. Borkird C, Choi JH, Sung ZR. Effect of 2,4-Dichlorophenoxyacetic acid on the expression of embryogenic program in carrot. Plant Physiol 1986;81:1143-6. https://doi.org/10.1104/pp.81.4.1143
  38. Fujimura T, Komamine A. Mode of action of 2,4-D and zeatin on somatic embryogenesis ina carrot cell suspensionculture.Z Pflanzenphysiol 1980;99:1-8. https://doi.org/10.1016/S0044-328X(80)80106-1
  39. Lippmann B, Lippmann G. Induction of somatic embryos in cotyledonary tissue of soybean, Glycine max L. Merr. Plant Cell Rep 1984;3:215-8. https://doi.org/10.1007/BF00269295
  40. Choi KT. Panax ginseng C. A. Meyer: micropropagation and the in vitro production of saponins. In: Bajaj YPS, editor. Medicinal and aromatic plants I. Berlin, Heidelberg: Springer; 1988. p. 484-500.
  41. Choi YE, Jeong JH, In JK, Yang DC. Production of herbicide-resistant transgenic Panax ginseng through the introduction of the phosphinothricin acetyl transferase gene and successful soil transfer. Plant Cell Rep 2003;21:563-8. https://doi.org/10.1007/s00299-002-0551-z
  42. Meijer EA, de Vries SC, Mordhorst AP. Co-culture with Daucus carota somatic embryos reveals high 2,4-D uptake and release rates of Arabidopsis thaliana cultured cells. Plant Cell Rep 1999;18:656-63. https://doi.org/10.1007/s002990050638
  43. Wernicke W, Milkovits L. Developmental gradients in wheat leaves - response of leaf segments in different genotypes cultured in vitro. J Plant Physiol 1984;115:49-58. https://doi.org/10.1016/S0176-1617(84)80050-4
  44. Mooney EH, McGraw JB. Effects of self-pollination and outcrossing with cultivated plants in small natural populations of American ginseng, Panax quinquefolius (Araliaceae). Am J Bot 2007;94:1677-87. https://doi.org/10.3732/ajb.94.10.1677
  45. Chen ZJ. Molecular mechanisms of polyploidy and hybrid vigor. Trends Plant Sci 2010;15:57. https://doi.org/10.1016/j.tplants.2009.12.003
  46. Brewbaker JL, Sun WG. Trees and heterosis. The genetics and exploitation of heterosis in crops: an international symposium. CIMMYT; 1997. p. 463-78.
  47. Yang DC, Choi YE. Production of transgenic plants via Agrobacterium rhizogenes-mediated transformation of Panax ginseng. Plant Cell Rep 2000;19:491-6. https://doi.org/10.1007/s002990050761
  48. Watanabe M, Hubberten H-M, Saito K, Hoefgen R. General regulatory patterns of plant mineral nutrient depletion as revealed by serat quadruple mutants disturbed in cysteine synthesis. Mol Plant 2010;3:438-66. https://doi.org/10.1093/mp/ssq009
  49. Ziv M. Transplanting Gladiolus plants propagated in vitro. Sci Hortic Amsterdam 1979;11:257-60. https://doi.org/10.1016/0304-4238(79)90008-6
  50. Konsler TR, Shelton JE. Lime and phosphorus effects on American ginseng: I. Growth, soil fertility, and root tissue nutrient status response. J Am Soc Hort Sci 1990;115:570-4. https://doi.org/10.21273/JASHS.115.4.570
  51. Bradford KJ, Hsiao TC. Physiological responses to moderate water stress. In: Lange OL, et al., editors. Physiological plant ecology II: water relations and carbon assimilation. Berlin, Heidelberg: Springer; 1982. p. 263-324.
  52. White CN, Proebsting WM, Hedden P, Rivin CJ. Gibberellins and seed development in maize. I. Evidence that gibberellin/abscisic acid balance governs germination versus maturation pathways. Plant Physiol 2000;122:1081-8. https://doi.org/10.1104/pp.122.4.1081
  53. Choi YE, Kim JW, Yoon ES. High frequency of plant production via somatic embryogenesis from callus or cell suspension cultures in Eleutherococcus senticosus. Ann Bot 1999;83:309-14. https://doi.org/10.1006/anbo.1998.0827
  54. Heun M, Helentjaris T. Inheritance of RAPDs in F1 hybrids of corn. Theor Appl Genet 1993;85:961-8. https://doi.org/10.1007/BF00215035
  55. Liu Z, Li P, Argue BJ, Dunham RA. Inheritance of RAPD markers in channel catfish (Ictalurus punctatus), blue catfish (I. furcatus), and their F1, F2 and backcross hybrids. Anim Genet 1998;29:58-62. https://doi.org/10.1046/j.1365-2052.1998.00284.x
  56. Gjuric R, Smith SR. Identification of cross-pollinated and self-pollinated progeny in Alfalfa through RAPD nulliplex loci analysis. Crop Sci 1996;36:389-93. https://doi.org/10.2135/cropsci1996.0011183X003600020029x
  57. Xu Y-S, Clark MS, Pehu E. Use of RAPD markers to screen somatic hybrids between Solanum tuberosum and S. brevidens. Plant Cell Rep 1993;12:107-9. https://doi.org/10.1007/BF00241944
  58. Verhaegen D, Plomion C, Gion J-M, Poitel M, Costa P, Kremer A. Quantitative trait dissection analysis in Eucalyptus using RAPD markers: 1. Detection of QTL in interspecific hybrid progeny, stability of QTL expression across different ages. Theor Appl Genet 1997;95:597-608. https://doi.org/10.1007/s001220050601
  59. Goldman JJ. The use of ISSR markers to identify Texas bluegrass interspecific hybrids. Plant Breeding 2008;127:644-6. https://doi.org/10.1111/j.1439-0523.2008.01526.x
  60. Hashemi SH, Mirmohammadi-Maibody SAM, Nematzadeh GA, Arzani A. Identification of rice hybrids using microsatellite and RAPD markers. Afr J Biotechnol 2009;8:2094-101.
  61. Noormohammadi Z, Shojaei-Jesvaghani F, Sheidai M, Farah F, Alishah O. Inter simple sequence repeats (ISSR) and random amplified polymorphic DNA (RAPD) analyses of genetic diversity in Mehr cotton cultivar and its crossing progenies. Afr J Biotechnol 2011;10:11839-47.
  62. Godwin ID, Aitken EAB, Smith LW. Application of inter simple sequence repeat (ISSR) markers to plant genetics. Electrophoresis 1997;18:1524-8. https://doi.org/10.1002/elps.1150180906
  63. Li TSC, Bedford KE, Sholberg PL. Improved germination of American ginseng seeds under controlled environments. HortTechnology 2000;10:131-5. https://doi.org/10.21273/HORTTECH.10.1.131
  64. Li TSC. Asian and American ginseng - a review. HortTechnology 1995;5:27-34. https://doi.org/10.21273/HORTTECH.5.1.27

Cited by

  1. Phalaenopsis LEAFY COTYLEDON1-Induced Somatic Embryonic Structures Are Morphologically Distinct From Protocorm-Like Bodies vol.10, 2019, https://doi.org/10.3389/fpls.2019.01594
  2. Application of genetics and biotechnology for improving medicinal plants vol.249, pp.4, 2019, https://doi.org/10.1007/s00425-019-03099-1
  3. Functional characterization of a WRKY family gene involved in somatic embryogenesis in Panax ginseng vol.257, pp.2, 2019, https://doi.org/10.1007/s00709-019-01455-2
  4. Major Achievement and Prospect of Ginseng Breeding in Korea vol.52, pp.1, 2019, https://doi.org/10.9787/kjbs.2020.52.s.170
  5. Genetic Composition of Korean Ginseng Germplasm by Collection Area and Resource Type vol.10, pp.11, 2019, https://doi.org/10.3390/agronomy10111643
  6. Overexpression of the squalene epoxidase gene (PgSE1) resulted in enhanced production of ginsenosides and phytosterols in transgenic ginseng vol.14, pp.6, 2019, https://doi.org/10.1007/s11816-020-00643-4
  7. Comparative Study of the Genetic and Biochemical Variability of Polyscias filicifolia (Araliaceae) Regenerants Obtained by Indirect and Direct Somatic Embryogenesis as a Source of Triterpenes vol.22, pp.11, 2021, https://doi.org/10.3390/ijms22115752