Cryopreservation of In Vitro Grown Apical Shoot Tips of Wasabia japonica (Wasabi) by Different Procedures

  • Toshikazu Matsumoto
Part of the Biotechnology in Agriculture and Forestry book series (AGRICULTURE, volume 50)


Wasabi, Japanese horseradish (family Cruciferae), is an important crop for Japanese foods such as sushi, sashimi and soba. The roots contain a pungent ingredient (sinigrin). Myrosinase is responsible for the development of the flavor and pungency of wasabi roots (Ohtsuru and Kawatani 1979). Wasabi also contains other useful compounds, such as peroxidase, that is widely used as a label enzyme in clinical diagnosis and immunoassay analysis (Taniguchi et al. 1988). The plant occurs in remote and inaccessible cool mountain springs and streams in Japan. The optimal growth temperature is about 15°C, and the growth is inhibited at high temperatures.


Sugar Crystallization Sucrose Glycerol DMSO 


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  1. Bapat VA, Mhatre M, Rao PS (1987) Propagation of Morus indica L. ( Mulberry) by encapsulated shoot buds. Plant Cell Rep 6: 393–395CrossRefGoogle Scholar
  2. Close TJ, Kortt AA, Chandler PM (1989) A cDNA-based comparison of dehydration-induced proteins (dehydrins) in barley and corn. Plant Mol Biol 13: 95–108PubMedCrossRefGoogle Scholar
  3. Crowe JH, Crowe JF, Carpenter LM, Wistrom CA (1987) Stabilization of dry phospholipid bilayers and proteins by sugars. Biochem J 242: 1–10PubMedGoogle Scholar
  4. Crowe JH, Crowe JF, Carpenter LM, Rudolph AS, Wistrom CA, Spargo BJ, Anchordoguy TJ (1988) Interaction of sugars with membranes. Biochem Biophys Acta 947: 367–384PubMedCrossRefGoogle Scholar
  5. Dereuddre J, Scottez C, Arnaud Y, Duron M (1990) Resistance of alginate-coated axillary shoot tips of pea tree (Pyrus communis L. cv. Beurre Hardy) in vitro plantlets to dehydration and subsequent freezing in liquid nitrogen: effects of previous cold hardening. CR Acad Sci Paris 310 (Ser I1I): 317–323Google Scholar
  6. Dereuddre J, Blandis S, Hassen N (1991) Resistance of alginate-coated somatic embryos of carrot (Daucus carota L.) to desiccation and freezing in liquid nitrogen. 1. Effects of preculture. Cryo Lett 12: 125–134Google Scholar
  7. Fabre J, Dereuddre J (1990) Encapsulation-dehydration: a new approach to cryopreservation of Solanum shoot tips. Cryo Lett 11: 413–426Google Scholar
  8. Fahy GM, MacFarlane DR, Angell CA, Meryman HT (1984) Vitrification as an approach to cryopreservation. Cryobiology 21: 407–426PubMedCrossRefGoogle Scholar
  9. Hirai D, Shirai K, Shirai S, Sakai A (1998) Cryopreservation of in vitro-grown shoot tips of strawberry by encapsulation/vitrification. Euphytica 101: 109–116CrossRefGoogle Scholar
  10. Jitsuyama Y, Suzuki T, Harada T, Fujikawa S (1997) Ultrastructural study on mechanism of increased freezing tolerance due to extracellular glucose in cabbage cells. Cryo Lett 18: 33–44Google Scholar
  11. Langis R, Schnabel-Preikstas BJ, Earle FD, Steponkus PL (1990) Cryopreservation of carnation shoot tips by vitrification. Cryobiology 27th Annual Meeting Abstract, pp 657–658Google Scholar
  12. Luo J, Reed B (1997) Abscisic acid-responsive protein, bovine serum albumin, and proline pre-treatments improve recovery of in vitro currant shoot-tip shoot tips and callus cryopreserved by vitrification. Cryobiology 34: 240–250CrossRefGoogle Scholar
  13. Matsumoto T (1997) Cryopreservation of in vitro-grown apical meristems of wasabi (Wasabia japonica). PhD Thesis, Department of Agronomy, Kobe University, KobeGoogle Scholar
  14. Matsumoto T, Nako Y (1999) Effect of dimethyl sulfoxide on in vitro storage of wasabi meristems at low temperature. Plant Biotech 16: 243–245CrossRefGoogle Scholar
  15. Matsumoto T, Sakai A (1995) An approach to enhance dehydration tolerance of alginate-coated dried meristems cooled to —196 °C. Cryo Lett 16: 299–306Google Scholar
  16. Matsumoto T, Sakai A (2000) Cryopreservation of grape in vitro-cultured axillary shoot tips by three-step vitrification. In: Engelmann F, Takagi H (eds) Cryopreservation of tropical germplasm. Current research progress and application. IPGRI, Rome, pp 266–267Google Scholar
  17. Matsumoto T, Sakai A, Yamada K (1994) Cryopreservation of in vitro-grown apical meristems of wasabi (Wasabia japonica) by vitrification and subsequent high plant regeneration. Plant Cell Rep 13: 442–446CrossRefGoogle Scholar
  18. Matsumoto T, Sakai A, Takahashi C, Yamada K (1995a) Cryopreservation of in vitro-grown apical meristems of wasabi (Wasabia japonica) by encapsulation-vitrification method. Cryo Lett 16: 189–206Google Scholar
  19. Matsumoto T, Sakai A, Yamada K (1995b) Cryopreservation of in vitro-grown apical meristems of lily (Lilium japonicum) by vitrification. Plant Cell Tissue Org Cul 41: 231–241Google Scholar
  20. Matsumoto T, Sakai A, Takahashi C, Yamada K (1996) Cryopreservation in vitro-grown apical meristems of lily (Lilium L.) by encapsulation-vitrification method. Plant Tissue Cult Lett 13: 29–34CrossRefGoogle Scholar
  21. Matsumoto T, Nako Y, Takahashi C, Sakai A (1997) Induction of in vitro cultured masses of shoot primordia of hybrid statice and its cryopreservation by vitrification. HortScience 32: 309–311Google Scholar
  22. Matsumoto T, Sakai A, Nako Y (1998a) A novel preculturing for enhancing the survival of in vitro-grown meristems of wasabi (Wasabia japonica) cooled to —196 °C by vitrification. Cryo Lett 19: 27–36Google Scholar
  23. Matsumoto T, Takahashi C, Sakai A, Nako Y (1998b) Cryopreservation of in vitro-grown apical meristems of hybrid statice by three different procedures. Sci Hort 76: 105–114CrossRefGoogle Scholar
  24. Matsumoto T, Mochda K, Itamura H (2000) Cryopreservation of dormant shoot tips of Diospyros by vitrification. Abstracts of the International Conference on Science and technology for managing plant genetic diversity in the 21st century, p 30Google Scholar
  25. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497CrossRefGoogle Scholar
  26. Niino T, Sakai A (1992) Cryopreservation of alginate-coated in vitro shoot tips of apple, pear and mulberry. Plant Sci 87: 199–206CrossRefGoogle Scholar
  27. Nishizawa S, Sakai A, Amano Y, Matsuzawa T (1992) Cryopreservation of asparagus (Asparagus officinalis L. Osb.) embryogenic cells and subsequent plant regeneration by a simple freezing method. Cryo Lett 13: 379–388Google Scholar
  28. 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: 67–73CrossRefGoogle Scholar
  29. Ohtsuru M, Kawatani H (1979) Studies on the myrosinase from Wasabia japonica: purification and some properties of wasabi myrosinase. Agric Biol Chem 43: 2249–2255CrossRefGoogle Scholar
  30. Phunchindawan M (1997) Cryopreservation of useful plant resources: Application of encapsulation-dehydration method to preservation of hairy root cultures and microalgae. PhD Thesis, Osaka University, Faculty of Pharmaceutical Sci, OsakaGoogle Scholar
  31. Reinhoud PJ, Schrijnemakers EWM, Iren F, Kijne JW (1995) Vitrification and a heat-shock treatment improve cryopreservation of tobacco cell suspension compared to two-step freezing. Plant Cell Tissue Org Cult 42: 261–267CrossRefGoogle Scholar
  32. Sakai A (1997) Potentially valuable cryogenic procedures for cryopreservation of cultured plant shoot tips. In: Razdan MK, Cocking EC (eds) Conservation of plant genetic resources in vitro. Science Publishers, New Hampshire, pp 53–66Google Scholar
  33. Sakai A, Yoshida S (1967) Survival of plant tissue at super-low temperature. Effects of cooling and rewarming rates on survival. Plant Physiol 42: 1695–1701PubMedCrossRefGoogle Scholar
  34. 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: 30–33CrossRefGoogle Scholar
  35. Sakai A, Kobayashi S, Oiyama I (1991) Cryopreservation of nucellar cells of navel orange (Citrus sinensis Osb.) by a simple freezing method. Plant Sci 74: 243–248CrossRefGoogle Scholar
  36. Skriver K, Mundy J (1990) Gene expression in response to abscisic acid and osmotic stress. Plant Cell 2: 503–512PubMedGoogle Scholar
  37. Steponkus PL, Langis R, Fujikawa S (1992) Cryopreservation of plant tissues by vitrification. In: Steponkus PL (ed) Advances in low temperature biology, vol 1. JAI Press, London, pp 1–61Google Scholar
  38. Suzuki M, Niino T, Akiyama T (1994) Cryopreservation of shoot tips of kiwifruit seedlings by the alginate encapsulation-dehydration technique. Plant Tissue Cult Lett 11: 122–128CrossRefGoogle Scholar
  39. Taniguchi M, Nomura R, Kamihara M, Kijima I, Kobayashi T (1988) Effective utilization of horseradish (Armoracia lapathifolia) and wasabi (Wasabia japonica) by treatment with supercritical carbon dioxide. J Ferment Bioeng 66: 347–353Google Scholar
  40. Thinh NT (1997) Cryopreservation of germplasm of vegetatively propagated tropical monocots by vitrification. PhD Thesis, Department of Agronomy, Kobe University, KobeGoogle Scholar
  41. Widholm JM (1972) The use of fluorescein diacetate and phenosafranine for determining viability of cultured plant cells. Stain Tech 47 (4): 189–194Google Scholar
  42. Yamada K, Haruki K (1992) Mass propagation of wasabi (Wasabia japonica Matsumura) through shoot apex culture. Bull Shimane Agric Exp Stn 26: 85–95Google Scholar
  43. Yoshimatsu K, Yamaguchi H, Shimomura K (1996) Traits of Panax ginseng hairy roots after cold storage and cryopreservation. Plant Cell Rep 15: 555–560CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2002

Authors and Affiliations

  • Toshikazu Matsumoto
    • 1
  1. 1.Shimane Agricultural Experiment StationIzumo, ShimaneJapan

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