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Development of PVS-Based Vitrification and Encapsulation–Vitrification Protocols

  • Akira Sakai
  • Dai Hirai
  • Takao Niino

Cryopreservation is a very important tool for the long-term storage of plant genetic resources for future generations, requiring only a minimum of space and maintenance. With increasing interest in the genetic engineering of plants, the preservation of cultured cells and somatic embryos with unique attributes is assuming greater importance. Recently, cryopreservation was reported to offer real hope for enhancing the preservation of endangered and rare plants (Touchell 1995; Touchell and Dixon 1996). This chapter describes protocols for successful vitrification using plant vitrification solution 2 (PVS2), and highlights some of the factors contributing to high levels of post-LN recovery. The development of a simple and reliable method for cryopreservation would allow the widespread storage of cultured cells, meristems, and somatic embryos. Vitrification involving vitrification solutions (Langis et al. 1990; Sakai et al. 1990; Yamada et al. 1991) and encapsulation-dehydration techniques (Fabre and Dereuddre 1990) were developed in the 1990s, and the number of cryopreserved species has increased markedly since then (Sakai 1995, 1997; Engelmann and Takagi 2000). A vitrification procedure using an ethylene glycol-based vitrification solution and French straws was presented by Steponkus and colleagues. They reported successful cryopreservation of Dianthus and Chrysanthemum (Langis et al. 1990; Schnabel-Preikstas et al. 1992), and potato (Lu and Steponkus 1994). This chapter will outline the development and uses of PVS2 developed by Sakai and associates.

Keywords

Somatic Embryo Hairy Root Apical Meristem Cold Acclimation Tropical Plant 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Agrawal A, Swennen R, Panis B (2004) A comparison of four methods for cryopreservation of meristems in banana (Musa spp.). CryoLetters 25: 101-110Google Scholar
  2. Burke MJ (1986) The glassy state and survival of anhydrous biological systems. In: Leopold AC (ed) Membrane, Metabolism and Dry Organisms. Cornell Univ. Press, Ithaca, NY, pp 358-364Google Scholar
  3. Charoensub R, Hirai D, Sakai A (2004) Cryopreservation of in vitro-grown shoot tips of cassava by encapsulation-vitrification method. CryoLetters 25: 51-58PubMedGoogle Scholar
  4. Charoensub R, Phansiri S, Sakai A, Yongmenitchai W (1999) Cryopreservation of cassava in vitro grown shoot tips cooled to -196°C by vitrification. CryoLetters 20: 89-94Google Scholar
  5. Chen Y, Wang JH (2003) Cryopreservation of carrot (Daucus carota L.) cell suspensions and protoplasts by vitrification. CryoLetters 24: 57-64PubMedGoogle Scholar
  6. Cho EG, Hor YL, Kim HH, Rao VR, Engelmann F (2002) Cryopreservation of Citrus mandurensis zygotic embryonic axes by vitrification: Importance of loading and treatment with vitrification solution. CryoLetters 23: 317-324PubMedGoogle Scholar
  7. Corredoira E, San-José MC, Ballester A, Vieitez AM (2004) Cryopreservation of zygotic embryo axes and somatic embryos of European chestnut. CryoLetters 25: 33-42PubMedGoogle Scholar
  8. Crowe JH, Crowe LF, Chapman D (1984a) Preservation of membranes in anhydrobiotic organisms: The role of trehalose. Science 233: 701-703Google Scholar
  9. Crowe JH, Crowe LF, Chapman D (1984b) Infrared spectroscopic studies on interactions of water and carbohydrates with a biological membrane. Arch Biochem Biophys 232: 400-407PubMedGoogle Scholar
  10. De Carlo A, Benelli C, Lambardi M (2000) Development of a shoot-tip vitrification protocol and comparison with encapsulation-based procedures for plum (Prunus domestica L.) cryopreservation. CryoLetters 21: 215-222PubMedGoogle Scholar
  11. Dereuddre J, Fabre J, Bassaglia C (1988) Resistance to freezing in liquid nitrogen of carnation (Dianthus caryophyllus L. var. Eolo) apical and axillary shoot tips excised from different aged in vitro plantlets. Plant Cell Rep 7: 170-173Google Scholar
  12. Dereuddre J, Blandin 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. CryoLetters 12: 125-134Google Scholar
  13. Dumet D, Engelmann F, Chabrillange N, Duval Y (1993a) Cryopreservation of oil palm (Elaeis guineensis Jacq.) somatic embryo involving a desiccation step. Plant Cell Rep 12: 352-355Google Scholar
  14. Dumet D, Engelmann F, Chabrillange N, Duval Y, Dereuddre J (1993b) Importance of sucrose for the acquisition of tolerance to desiccation and cryopreservation of oil palm somatic embryos. CryoLetters 14: 243-250Google Scholar
  15. Engelmann F (1991) In vitro conservation of tropical plant germplasm - a review. Euphytica 57: 227-243Google Scholar
  16. Engelmann F, Takagi H (2000) Cryopreservation of Tropical Plant Germplasm: Current Research Progress and Applications. JIRCAS, Tsukuba and IPGRI, RomeGoogle Scholar
  17. Fabre J, Dereuddre J (1990) Encapsulation-dehydration: A new approach to cryo-preservation of Solanum shoot tips. CryoLetters 11: 413-426Google Scholar
  18. Fahy GM, MacFarlane DR, Angell CA, Meryman HT (1984) Vitrification as an approach to cryopreservation. Cryobiology 21: 407-426PubMedGoogle Scholar
  19. Fukai S (1992) Studies on the cryopreservation of shoot tips of Dianthus and Chrysanthemum. Mem Fac Agric Kagawa Univ 56: 1-79Google Scholar
  20. Gagliardi RF, Pacheco GP, Cameiro LA, Valls JFM, Vieira MLC, Mansur E (2003) Cryopreservation of Arachis species by vitrification of in vitro grown shoot apices and genetic stability of recovered plants. CryoLetters 24: 103-110PubMedGoogle Scholar
  21. Gamez-Pastrana R, Martinez-Ocampo Y, Beristain CI, Gonzalez-Arnao MT (2004) An improved cryopreservation protocol for pineapple apices using encapsulation-vitrification. CryoLetters 25: 405-414PubMedGoogle Scholar
  22. Golmirzaie AM, Panta A (2000) Advances in potato cryopreservation at CIP. In: Engelmann F, Takagi H (eds) Cryopreservation of Tropical Plant Germplasm: Current Research Progress and Applications. JIRCAS, Tsukuba and IPGRI, Rome. pp 250-254Google Scholar
  23. Gonzalez-Arnao MT, Ravelo MM, Villavicencio CU, Montero MM, Engelmann F (1998) Cryopreservation of pineapple (Ananas comosus) apices. CryoLetters 19: 375-382Google Scholar
  24. Hirai D, Sakai A (1999a) Cryopreservation of in vitro grown axillary shoot-tip meristems of mint (Mentha spicata) by encapsulation-vitrification. Plant Cell Rep 19: 150-155Google Scholar
  25. Hirai D, Sakai A (1999b) Cryopreservation of in vitro grown meristems of potato (Solanum tuberosum L.) by encapsulation-vitrification. Potato Res 42: 153-160Google Scholar
  26. Hirai D, Sakai A (2003) Simplified cryopreservation of sweet potato (Ipomoea batatas (L.) Lam.) by optimizing conditions for osmoprotection. Plant Cell Rep 21: 961-966PubMedGoogle Scholar
  27. Hirai D, Shirai K, Shirai S, Sakai A (1998) Cryopreservation of in vitro-grown meristems of strawberry (Fragaria x ananassa Duch.) by encapsulationvitrification. Euphytica 101: 109-115Google Scholar
  28. Huang CN, Wang JH, Yan QS, Zhang XQ, Yan QF (1995) Plant regeneration from rice (Oryza sativa L.) embryogenic suspension cells cryopreserved by vitrification. Plant Cell Rep 14: 730-734Google Scholar
  29. Ishikawa K, Harada K, Mii M, Sakai A, Yoshimatsu K, Shimomura K (1997) Cryopreservation of protocorms of Japanese terrestrial orchid (Bletilla striata) by vitrification. Plant Cell Rep 16: 754-757Google Scholar
  30. Jung DW, Sung CK, Touno K, Yoshimatsu K, Shimomura K (2001) Cryopreservation of Hyoscyamus niger adventitious roots by vitrification. J Plant Physiol 158: 801-805Google Scholar
  31. Kamiya T, Sasaki K, Yoshimatsu K, Shimomura K (1995) Cryopreservation of hairy roots of Atropa belladona by vitrification. Proc Japan Soc Plant Tissue Cult., 14th Annual Meet Rep:112Google Scholar
  32. Kim HH (2004) Analysis of factors affecting the cryopreservation of garlic shoot tips. Doctoral Paper, Graduate School, Seoul National University, KoreaGoogle Scholar
  33. Kim HH, Cho EG, Baek YJ, Kim CY, Engelmann F (2004) Cryopreservation of garlic shoot tips by vitrification: effect of dehydration, rewarming, unloading and regrowth conditions. CryoLetters 25: 59-70PubMedGoogle Scholar
  34. Kohmura H, Ikeda Y, Sakai A (1994) Cryopreservation of apical meristems of Japanese shallot (Allium wakegi) by vitrification and subsequent high plant regeneration. CryoLetters 15: 289-298Google Scholar
  35. Kohmura H, Sakai A, Chokyu S, Yakuwa T (1992) Cryopreservation of in vitrocultured multiple bud clusters of asparagus (Asparagus officinalis L. cv. Hiroshima green) by the technique of vitrification. Plant Cell Rep 11: 433-437Google Scholar
  36. Kuranuki Y, Sakai A (1995) Cryopreservation of in vitro-grown shoot tips of tea (Camellia sinensis) by vitrification. CryoLetters 16: 345-352Google Scholar
  37. Lambardi M, Fabbri A, Caccavale A (2000) Cryopreservation of white poplar (Populus alba L.) by vitrification of in vitro grown shoot tips. Plant Cell Rep 19: 213-218Google Scholar
  38. Langis R, Schnabel B, Earle ED, Steponkus PL (1989) Cryopreservation of Brassica campestris cell suspensions by vitrification. CryoLetters 10: 421-428Google Scholar
  39. Langis R, Schnabel-Preikstas BJ, Earle ED, Steponkus PL (1990) Cryopreservation of carnation shoot tips by vitrification. Cryobiology 276: 657-658Google Scholar
  40. Leunufna S, Keller ERJ (2005) Cryopreservation of yams using vitrification modified by including droplet method: Effects of cold acclimation and sucrose. CryoLetters 26: 93-101PubMedGoogle Scholar
  41. Langis R, Steponkus P (1991) Vitrification of isolated rye protoplasts: protection against dehydration injury by ethylene glycol. CryoLetters 12: 107-112Google Scholar
  42. Lu S, Steponkus PL (1994) Cryopreservation of Solanum shoot-tips by vitrification. Cryobiology 31: 569Google Scholar
  43. Luo J, Reed BM (1997) Abscisic acid-responsive protein, bovine serum albumin, and proline pretreatments improve recovery of in vitro currant shoot-tip meristems and callus cryopreserved by vitrification. Cryobiology 34: 240-250Google Scholar
  44. Luyet BJ (1937) The vitrification of organic colloids and protoplasm. Biodinamica 29: 1-15Google Scholar
  45. Makowska Z, Keller J, Engelmann F (1999) Cryopreservation of apices isolated from garlic (Allium sativum L.) bulbils and cloves. CryoLetters 20: 175-182Google Scholar
  46. Martinez MT, Ballester A, Vieitez AM (2003) Cryopreservation of embryogenic cultures of Quercus robur using desiccation and vitrification procedures. Cryobiology 46: 182-189PubMedGoogle Scholar
  47. Matsumoto T, Mochida K, Itamura H, Sakai A (2001) Cryopreservation of persimmon (Diopsyros kaki Thunb.) by vitrification of dormant shoot tips. Plant Cell Rep 20: 398-402Google Scholar
  48. Matsumoto T, Sakai A (2003) Cryopreservation of axillary shoot tips of in vitrogrown grape (Vitis) by a two-step vitrification protocol. Euphytica 131: 299-304Google Scholar
  49. Matsumoto T, Sakai A, Nako Y (1998) A novel preculturing for enhancing the survival of in vitro grown meristems of wasabi (Wasabia japonica) cooled to -196°C by vitrification. CryoLetters 19: 27-36Google Scholar
  50. Matsumoto T, Sakai A, Takahashi C, Yamada K (1995a) Cryopreservation of in vitro-grown apical meristems of wasabi (Wasabia japonica) by encapsulation-vitrification method. CryoLetters 16: 189-196Google Scholar
  51. 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: 442-446Google Scholar
  52. Matsumoto T, Sakai A, Yamada K (1995b) Cryopreservation of in vitro-grown apical meristems of lily by vitrification. Plant Cell Tissue Organ Cult 41: 237-241Google Scholar
  53. Matsumoto T, Takahashi C, Sakai A, Nako Y (1998) Cryopreservation of in vitrogrown apical meristems of hybrid statice by three different procedures. Scientia Hortic 76: 105-114Google Scholar
  54. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15: 473-497Google Scholar
  55. Niino T, Hettiarachchi A, Takahashi J, Samarajeewa PK (2000a) Cryopreservation of lateral buds of in vitro-grown plants of innala (Solemostemon rotundifolius) by vitrification. CryoLetters 21: 349-356PubMedGoogle Scholar
  56. Niino T, Ivette S, Murayama T (2000b) Cryopreservation of vegetatively propagated species (mainly mulberry). In: Engelmann F, Takagi H (eds) Cryopreservation of Tropical Plant Germplasm: Current Research Progress and Applications. JIRCAS, Tsukuba and IPGRI, Rome, pp 75-80Google Scholar
  57. Niino T, Sakai A, Yakuwa H, Nojiri K (1992a) Cryopreservation of in vitrogrown shoot tips of apple and pear by vitrification. Plant Cell Tissue Organ Cult 28: 261-266Google Scholar
  58. Niino T, Sakai A, Enomoto S, Magoshi J, Kato S (1992b) Cryopreservation of in vitro-grown shoot tips of mulberry by vitrification. CryoLetters 13: 303-312Google Scholar
  59. Niino T, Takano J, Saga T, Kobayashi M (2003a) Cryopreservation of in vitro grown Chinese leek “Nakamidori” shoot tips by vitrification. Hortic Res Jpn 2: 241-245Google Scholar
  60. Niino T, Tanaka D, Ichikawa S, Takano J, Ivette S, Shirata K, Uemura M (2003b) Cryopreservation of in vitro-grown apical shoot tips of strawberry by vitrification. Plant Biotech 20: 75-80Google Scholar
  61. Niino T, Tashiro K, Suzuki M, Ohuchi S, Magoshi J, Akihama T (1997) Cryopreservation of in vitro-grown shoot tips of cherry and sweet cherry by one-step vitrification. Scientia Hortic 70: 155-163Google Scholar
  62. Nishizawa S, Sakai A, Amano Y, Matsuzawa T (1992) Cryopreservation of asparagus (Asparagus officinalis L.) embryogenic suspension cells and subsequent plant regeneration by a simple freezing method. CryoLetters 13: 379-388Google Scholar
  63. 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: 67-73Google Scholar
  64. Niwata E (1995) Cryopreservation of apical meristems of garlic (Allium sativum L.) and high subsequent plant regeneration. CryoLetters 16: 102-107Google Scholar
  65. Panis B (1995) Cryopreservation of banana (Musa spp.) germplasm. Doctoral paper (No. 272), Katholieke Universiteit, LeuvenGoogle Scholar
  66. Paul H, Daigny G, Sangwan-Noreel BS (2000) Cryopreservation of apple (Malus x domestica Borkh.) shoot tips following encapsulation or encapsulationvitrification. Plant Cell Rep 19: 768-774Google Scholar
  67. Pennycooke JC, Towill LE (2001) Medium alterations improve regrowth of sweet potato (Ipomoea batatas (L.) Lam.) shoot tips by vitrification and encapsulation-dehydration. CryoLetters 22: 381-389PubMedGoogle Scholar
  68. Phunchindawan M, Hirata K, Miyamoto K, Sakai A (1997) Cryopreservation of encapsulated shoot primordia induced by horseradish (Armoracia rusticana) hairy roots. Plant Cell Rep 16: 469-473Google Scholar
  69. Reed BM (1992) Cryopreservation of Ribes apical meristems. Cryobiology 29: 740Google Scholar
  70. Reinhoud P (1996) Cryopreservation of tobacco suspension cells by vitrification. Doctoral Paper, Rijks University, LeidenGoogle Scholar
  71. Sakai A (1960) Survival of the twig of woody plants. Nature 185: 393-394Google Scholar
  72. Sakai A (1995) Cryopreservation of germplasm of woody plants. In: Bajaj YPS (ed) Biotechnology in Agriculture and Forestry, Vol. 32, Cryopreservation of Plant Germplasm I , Springer-Verlag, Berlin, Heidelberg, pp 53-69Google Scholar
  73. Sakai A (1997) Potentially valuable cryogenic procedures for cryopreservation of cultured plant meristems. In: Razdan MK, Cocking EC (eds) Conservation of Plant Genetic Resources In Vitro Science Publishers, USA, pp 53-66Google Scholar
  74. 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: 30-33Google Scholar
  75. Sakai A, Kobayashi S, Oiyama I (1991a) Cryopreservation of nucellar cells of navel orange (Citrus sinensis Osb.) by a simple freezing method. Plant Sci 74: 243-248Google Scholar
  76. Sakai A, Kobayashi S, Oiyama I (1991b) Survival by vitrification of nucellar cells of navel orange (Citrus sinensis var. brasiliensis Tanaka) cooled to -196°C. J. Plant Physiol 137: 465-470Google Scholar
  77. Sakai A, Nishiyama Y (1978) Cryopreservation of winter vegetative buds of hardy fruit trees in liquid nitrogen. HortScience 13: 225-227Google Scholar
  78. Sakai A, Yoshida S (1967) Survival of plant tissue at super-low temperatures. VI. Effects of cooling and rewarming rates on survival. Plant Physiol 42: 1695-1701Google Scholar
  79. Schnabel-Preikstas B, Earle ED, Steponkus PL (1992) Cryopreservation of chrysanthemum shoot tips by vitrification. Cryobiology 29: 739Google Scholar
  80. Sharma N, Sharma B (2003) Cryopreservation of shoot tips of Picrorhiza kurroa Royle ex Benth., an indigenous endangered medicinal plant, through vitrification. CryoLetters 24: 181-190PubMedGoogle Scholar
  81. Shatnawi MA, Engelmann F, Frattarelli A, Damiano C (1999) Cryopreservation of apices of in vitro plantlets of almond (Prunus dulcis Mill). CryoLetters 20: 13-20Google Scholar
  82. Shibli RA, Al-Juboory KH (2000) Cryopreservation of “nabali” olive (Olea europaea L.) somatic embryos by encapsulation-dehydration and encapsulationvitrification. CryoLetters 21: 357-366PubMedGoogle Scholar
  83. Steponkus PL, Langis R, Fujikawa S (1992) Cryopreservation of plant tissues by vitrification. In: Steponkus PL (ed), Advances in Low Temperature Biology V.1. JAI Press, Hampton Hill, UK, pp 1-61Google Scholar
  84. Tanaka D, Niino T, Isuzugawa K, Hikage T, Uemura M (2004) Cryopreservation of shoot apices of in vitro grown gentian plants: comparison of vitrification and encapsulation-vitrification protocols. CryoLetters 25: 167-176PubMedGoogle Scholar
  85. Takagi H, Thinh NT, Kyesmu PM (1998) Cryopreservation of vegetatively propagated tropical crops by vitrification. Acta Hortic 461: 485-494Google Scholar
  86. Takagi H, Thinh NT, Sakai A, Senboku T (1997) Cryopreservation of taro in vitrogrown shoot tips (Colocasia esculenta (L.) Schott) by vitrification. I. Investigation of basic vitrification conditions. Plant Cell Rep 16: 594-599Google Scholar
  87. Thammasiri K (1999) Cryopreservation of embryonic axes of jackfruit. CryoLetters 20: 21-28Google Scholar
  88. Thammasiri K (2000) Cryopreservation of seeds of a Thai orchid (Doritis pulcherrima Lindl.) by vitrification. CryoLetters 21: 237-244PubMedGoogle Scholar
  89. Thinh NT (1997) Cryopreservation of germplasm of vegetatively propagated tropical monocots by vitrification. Doctoral Paper, Faculty of Agriculture, Kobe University, JapanGoogle Scholar
  90. Thinh NT, Takagi H (2000) Cryopreservation of in vitro grown apical meristems of terrestrial orchids (Cymbidium spp.) by vitrification. In: Engelmann F, Takagi H (eds) Cryopreservation of Tropical Plant Germplasm: Current Research Progress and Applications. JIRCAS, Tsukuba and IPGRI, Rome, pp 441-443Google Scholar
  91. Thinh NT, Takagi H, Sakai A (2000) Cryopreservation of in vitro grown apical meristems of some vegetatively propagated tropical monocots by vitrification. In: Engelmann F, Takagi H (eds) Cryopreservation of Tropical Plant Germplasm: Current Research Progress and Applications. JIRCAS, Tsukuba and IPGRI, Rome, pp 227-232Google Scholar
  92. Thinh NT, Takagi H, Yashima S (1999) Cryopreservation of in vitro grown shoot tips of banana (Musa spp.) by vitrification method. CryoLetters 20: 163-175Google Scholar
  93. Touchell D (1995) Principles of cryobiology for conservation of threatened Aus-tralian plants. Doctoral paper, Department of Botany, University of Western Australia, Perth.Google Scholar
  94. Touchell DH, Dixon KW (1996) Cryopreservation for the conservation of Australian endangered plants. In: Normah MN, Narimah MK, Clyde MM (eds) International Workshop on the In Vitro Conservation of Plant Genetic Resources, Plant Bio-technology Lab., University Kebangsaan Malaysia, Kuala Lumpur, pp 169-180Google Scholar
  95. Towill LE (1990) Cryopreservation of isolated mint shoot tips by vitrification. Plant Cell Rep 9: 178-180Google Scholar
  96. Towill LE, Bonnart R (2003) Cracking in a vitrification solution during cooling or warming does not affect growth of cryopreserved mint shoot tips. CryoLetters 24: 341-346PubMedGoogle Scholar
  97. Towill LE, Jarret RL (1992) Cryopreservation of sweet potato (Ipomoea batatas (L.) Lam.) shoot tips by vitrification. Plant Cell Rep 11: 175-178Google Scholar
  98. Tsukazaki H, Mii M, Tokuhara K, Ishikawa K (2000) Cryopreservation of Doritaenopsis suspension culture by vitrification. Plant Cell Rep 19: 1160-1164Google Scholar
  99. Turner SR, Tan B, Senaratna T, Bunn E, Dixon KW, Touchell DH (2000) Cryopreservation of the Australian species Macropidia fuliginosa (haemo-doraceae) by vitrification. CryoLetters 21: 379-388PubMedGoogle Scholar
  100. Uragami A, Sakai A, Nagai M (1990) Cryopreservation of dried axillary buds from plantlets of Asparagus officinalis L. grown in vitro. Plant Cell Rep 9: 328-331Google Scholar
  101. Valladares S, Toribio M, Celestino C, Vietiez A (2004) Cryopreservation of embryogenic cultures from mature Quercus suber trees using vitrification. CryoLetters 25: 177-186PubMedGoogle Scholar
  102. Vandenbussche B, Weyens G, De Proft MP (2000) Cryopreservation of in vitro sugar beet (Beta vulgaris L.) shoot tips by a vitrification technique. Plant Cell Rep 19: 1064-1068Google Scholar
  103. Volk GM, Maness N, Rotindo K (2004) Cryopreservation of garlic (Allium sativum L.) using plant vitrification solution 2. CryoLetters 25: 219-226PubMedGoogle Scholar
  104. Wang Q, Batuman Ö, Li P, Bar-Joseph M, Gafny R (2002) A simple and efficient cryopreservation of in vitro-grown shoot tips of ‘Troyer’ citrange [Poncirus trifoliata (L.) Raf. x Citrus sinensis (L.) Osbeck.] by encapsulation-vitrification. Euphytica 128: 135-142Google Scholar
  105. Wang ZC, Deng XX (2004) Cryopreservation of shoot-tips of citrus using vitrification: effect of reduced form of glutathione. CryoLetters 25: 43-50PubMedGoogle Scholar
  106. Watanabe K, Steponkus PL (1995) Vitrification of Oryza sativa L. cell suspensions. CryoLetters 16: 255-262Google Scholar
  107. Withers LA (1979) Freeze preservation of somatic embryos and clonal plantlets of carrot (Daucus carota). Plant Physiol 63: 460-467PubMedGoogle Scholar
  108. Wu Y, Zhao Y, Engelmann F, Zhou M, Zang D, Chen S (2001) Cryopreservation of apple dormant buds and shoot tips. CryoLetters 22: 375-380PubMedGoogle Scholar
  109. Wu Y, Engelmann F, Zhao Y, Zhou M, Chen S (1999) Cryopreservation of apple shoot tips: importance of cryopreservation technique and of conditioning of donor plants. CryoLetters 20: 121-130Google Scholar
  110. Yamada T, Sakai A, Matsumura T, Higuchi S (1991) Cryopreservation of apical meristems of white clover (Trifolium repens L.) by vitrification. Plant Sci 78: 81-87Google Scholar
  111. Yoshimatsu K, Yamaguchi H, Shimomura K (1996) Traits of Panax ginseng hairy roots after cold storage and cryopreservation. Plant Cell Rep 15: 555-560Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Akira Sakai
    • 1
  • Dai Hirai
    • 2
  • Takao Niino
    • 3
  1. 1.Hokkaido UniversityKitakuJapan
  2. 2.Hokkaido Central Agricultural Experiment StationNaganumaJapan
  3. 3.National Institute of Agrobiological SciencesKannondaiJapan

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