Abstract
We tested desiccation and/or vitrification procedures to cryopreserve the adventitious roots of Panax ginseng, the source of commercially produced ginsenosides. When only desiccation was applied, the post-freeze survival of 3- to 4-mm root tips was <14% regardless of the composition of the preculture medium or the explant origin. Callus formation was frequently observed after cryopreservation. In contrast, 90% survival and 32.5% root formation efficiency were achieved after cryopreservation when a vitrification protocol was followed. Adventitious root cultures in flasks and bioreactors were reestablished from root tips cryopreserved by vitrification. A prolonged lag-phase and lower biomass production were recorded in post-freeze-regenerated cultures compared with control roots that were subcultured four times in flasks. However, biomass accumulations did not differ between control and regenerated roots at the end of the sixth subculturing period. After 40 days of culture in bioreactors, a mean value of 12.5 g dw L−1 was recorded for post-freeze-regenerated cultures versus 9.1 g dw L−1 for the control roots. Production of triol and diol ginsenosides in our bioreactor cultures also was enhanced after cryopreservation, by 41.0% and 89.8%, respectively. These results suggest that the vitrification method is successful for cryopreservation of P. ginseng adventitious roots.
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References
Benson EE, Hamill JD (1991) Cryopreservation and post freeze molecular and biosynthetic stability in transformed roots of Beta vulgaris and Nicotiana rustica. Plant Cell Tiss Organ Cult 24:163–172
Bomal C, Tremblay FM (2000) Dried cryopreserved somatic embryos of two Picea species provide suitable material for direct plantlet regeneration and germplasm storage. Ann Bot 86:177–183
Chen CF, Chiou WF, Zhang JT (2008) Comparison of the pharmacological affects of Panax ginseng and Panax quinquefolium. Acta Pharmacol Sin 29:1103–1108
Choi YE, Kim YS, Yi MJ, Park WG, Yi JS, Chun SR, Han SS, Lee SJ (2007) Physiological and chemical characteristics of field- and mountain-cultivated ginseng roots. J Plant Biol 50:198–205
Danso KE, Ford-Lloyd BV (2004) Cryopreservation of embryogenic calli of cassava using sucrose cryoprotection and air desiccation. Plant Cell Rep 22:623–631
Engelmann F (2004) Plant cryopreservation: progress and prospects. In Vitro Cell Dev. Biol Plant 40:427–433
Grenier-de March G, de Boucaud MT, Chmielarz P (2005) Cryopreservation of Prunus avium L. embryogenic tissues. Cryo-Lett 26:341–348
Hahn EJ, Kim YS, Yu KW, Jeong CS, Paek KY (2003) Adventitious root cultures of Panax ginseng C.A. Meyer and ginsenoside production through large-scale bioreactor system. J Plant Biotechnol 5:1–6
Hirata K, Goda S, Phunchindawan M, Du D, Ishio M, Sakai A, Miyamoto K (1998) Cryopreservation of horseradish hairy root cultures by encapsulation-dehydration. J Ferment Bioeng 86:418–420
Hirata K, Mukai M, Goda S, Ishio-Kinugasa M, Yoshida K, Sakai A, Miyamoto K (2002) Cryopreservation of hairy root cultures of Vinca minor (L.) by encapsulation-dehydration. Biotechnol Lett 24:371–376
Joshi A, Teng WL (2000) Cryopreservation of Panax ginseng cells. Plant Cell Rep 19:971–977
Jung DW, Sung CK, Touno K, Yoshimatsu K, Shimomura K (2001) Cryopreservation of Hyoscyamus niger adventitious roots by vitrification. J Plant Physiol 158:801–805
Kartha KK, Engelmann F (1994) Cryopreservation and germplasm storage. In: Indra KV, Thorpe TA (eds) Plant Cell and Tissue Culture. Kluwer, Dordrecht, pp 195–230
Kim YS, Yeung EC, Hahn EJ, Paek KY (2007) Combined effects of phytohormone, indole-3-butyric acid, and methyl jasmonate on root growth and ginsenoside production in adventitious root cultures of Panax ginseng C.A. Meyer. Biotechnol Lett 29:1789–1792
Lambert E, Geelen D (2007) Cryopreservation of hairy root cultures from Maesa lanceolata. Commun Agric Appl Biol Sci 72:225–228
Lambert E, Goossens A, Panis B, van Labeke MC, Geelen D (2009) Cryopreservation of hairy root cultures of Maesa lanceolata and Medicago truncatula. Plant Cell Tiss Organ Cult 96:289–296
Nishizawa S, Sakai A, Amano Y, Matsuzawa T (1993) Cryopreservation of asparagus (Asparagus officinalis L.) embryogenic suspension cells and subsequent plant regeneration by vitrification. Plant Sci 91:67–73
Sakai A (2000) Development of cryopreservation techniques. In: Engelmann F, Takagi H (eds) Cryopreservation of Tropical Plant Germplasm: Current Research Progress and Applications. JIRCAS, IPGRI, Rome, pp 1–7
Sakai A, Engelmann F (2007) Vitrification, encapsulation-vitrification and droplet-vitrification: a review. Cryo-Lett 28:151–172
Sakai A, Kobayashi S, Oiyama I (1990) Cryopreservation of nucellar cells of navel orange (Citrus sinensis Osb. var. brasiliensis Tanaka) by vitrification. Plant Cell Rep 9:30–33
Teoh KH, Weathers PJ, Cheetham RD, Walcerz DB (1996) Cryopreservation of transformed (hairy) roots of Artemisia annua. Cryobiology 33:106–117
Touno K, Yoshimatsu K, Shimomura K (2006) Characteristics of Atropa belladonna hairy roots cryopreserved by vitrification method. Cryo-Lett 27:65–72
Wu CH, Tewari RK, Hahn EJ, Paek KY (2007) Nitric oxide elicitation induces the accumulation of secondary metabolites and antioxidant defense in adventitious roots of Echinacea purpurea. J Plant Biol 50:636–643
Xue SH, Luo XJ, Wu ZH, Zhang HL, Wang XY (2008) Cold storage and cryopreservation of hairy root cultures of medicinal plant Eruca sativa Mill., Astragalus membranaceus and Gentiana macrophylla Pall. Plant Cell Tiss Organ Cult 92:251–260
Yoshimatsu K, Yamaguchi H, Shimomura K (1996) Traits of Panax ginseng hairy roots after cold storage and cryopreservation. Plant Cell Rep 15:555–560
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This work was supported by a Korea Science and Engineering Foundation grant funded by the Korean Government (MOST; R01-2007-000-10543-0).
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Oh, S.Y., Wu, C.H., Popova, E. et al. Cryopreservation of Panax ginseng Adventitious Roots. J. Plant Biol. 52, 348–354 (2009). https://doi.org/10.1007/s12374-009-9045-7
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DOI: https://doi.org/10.1007/s12374-009-9045-7