Skip to main content
SpringerLink
Log in

Cryopreservation of immature seeds of Bletilla striata by vitrification

  • Cell Biology and Morphogenesis
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

An efficient protocol was established for the cryopreservation of immature seeds of a terrestrial orchid, Bletilla striata. Immature seeds collected 2–4 months after pollination (MAP) were treated using three different cryogenic procedures: (1) direct plunging into liquid nitrogen, (2) vitrification, and (3) vitrification with preculture. When immature seeds collected 3 MAP and 4 MAP were precultured for 3 days on New Dogashima medium supplemented with 0.3 M sucrose and cryopreserved by vitrification, the survival rate after preservation, as assessed by staining with 2,3,5-triphenyltetrazolium chloride, was 92% and 81%, respectively. Immature seeds thus treated showed no decrease in germination rate relative to untreated immature seeds, and they developed into normal plantlets in vitro.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Abbreviations

MAP :

Month(s) after pollination

ND :

New Dogashima

TTC :

2,3,5-Triphenyltetrazolium chloride

References

  • Buchheim JA, Colbum SM, Ranch JP (1989) Maturation of soybean somatic embryos and transition to plantlet growth. Plant Physiol 89:768–775

    Google Scholar 

  • Golovina EA, Hoekstra FA, Van Aelst CV (2001) The competence to acquire cellular desiccation tolerance is independent of seed morphological development. J Exp Bot 52:1015–1027

    Article  CAS  PubMed  Google Scholar 

  • Ishikawa K, Harata K, Mii M, Sakai A, Yoshimatsu K, Shimomura K (1997) Cryopreservation of zygotic embryos of Japanese terrestrial orchid (Bletilla striata) by vitrification. Plant Cell Rep 16:754–757

    Article  CAS  Google Scholar 

  • Kanchit T (2000) Cryopreservation of seeds of a Thai orchid (Doritis pulcherrima Lindl.) by vitrification. Cryo Lett 21:237–244

    Google Scholar 

  • Kohmura H, Sakai A, Chokyu S, Yakuwa T (1992) Cryopreservation of in vitro-cultured multiple bud clusters of asparagus [Asparagus officinalis L. cv. Hiroshimagreen (2n=30)] by the techniques of vitrification. Plant Cell Rep 11:433–437

    Article  Google Scholar 

  • Larkin PJ, Scowcroft WR (1981) Somaclonal variation—a novel source of variability from cell cultures for plant improvement. Theor Appl Genet 60:197–214

    Google Scholar 

  • Manning JC, Van Staden J (1987) The development and mobilization of seed reserves in some African orchids. Aust J Bot 35:343–353

    CAS  Google Scholar 

  • Miyoshi K, Mii M (1988) Ultrasonic treatment for enhancing seed germination of terrestrial orchid, Calanthe discolor, in asymbiotic culture. Sci Hortic 35:127–130

    Article  Google Scholar 

  • Miyoshi K, Mii M (1998) Stimulatory effects of sodium and calcium hypochlorite, pre-chilling and cytokinins on germination of Cypripedium macranthos seed in vitro. Physiol Plant 102:481–486

    Article  CAS  Google Scholar 

  • Nagashima T (1982a) Studies on the seed germination and embryogenesis in the Bletilla striata Rchb. f. and Calanthe discolor Lindl. J Jpn Soc Hortic Sci 51:82–93

    Google Scholar 

  • Nagashima T (1982b) Studies on the seed germination and embryogenesis in the Cymbidium goeringii Rchb. f. and Paphiopedilum insigne var. sanderae Rchb. f. J Jpn Soc Hortic Sci 51:94–105

    Google Scholar 

  • Nishizawa S, Sakai A, Amano Y, Matuzawa T (1993) Cryopreservation of asparagus (Asparagus officinalis L.) embryogenic suspension cells and subsequent plant regeneration by vitrification. Plant Sci 91:67–73

    Article  CAS  Google Scholar 

  • Pence VC (1992) Desiccation and the survival of Aesculus, Castanea, and Quercus embryo axes through cryopreservation. Cryobiology 29:391–399

    Google Scholar 

  • Rasmussen HN (1995) Cultivation of immature seed. In: Rasmussen HN (ed) Terrestrial orchids from seed to mycotrophyic plant. Cambridge University Press, Cambridge, UK, pp 17–38

    Google Scholar 

  • Roos EE, Davidson DA (1992) Record longevities of vegetable seeds in storage. Hortic Sci 27:393–396

    Google Scholar 

  • 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

    Article  Google Scholar 

  • Tokuhara K, Mii M (1993) Micropropagation of Phalaenopsis and Doritaenopsis by culturing shoot tips of flower stalk buds. Plant Cell Rep 13:7–11

    Article  CAS  Google Scholar 

  • Tsukazaki H, Mii M, Tokuhara K, Ishikawa K (2000) Cryopreservation of Doritaenopsis suspension culture by vitrification. Plant Cell Rep 19:1160–1164

    Article  CAS  Google Scholar 

  • Uragami A, Sakai A, Nagai M, Takahashi T (1989) Survival of cultured cells and somatic embryos of Asparagus officinalis cryopreserved by vitrification. Plant Cell Rep 8:418–421

    Article  Google Scholar 

  • Wang JH, Ge JG, Liu F, Bian HW, Huang CN (1998) Cryopreservation of seeds and protocorms of Dendrobium candidum. Cryo Lett 19:123–128

    Google Scholar 

  • Yamada T, Sakai A, Matsumura T, Higuchi S (1991) Cryopreservation of apical meristems of white clover (Trifolium repens L.) by vitrification. Plant Sci 78:81–87

    Article  CAS  Google Scholar 

  • Yeung EC, Law SK (1992) Embryology of Calypso bulbosa. II Embryo development. Can J Bot 70:461–468

    Google Scholar 

Download references

Author information

Author notes

  1. K. Ishikawa

    Present address: Japan Horticultural Production and Research Institute, 2-5-1 Kamishiki, Matsudo-City, Chiba, 270-2221, Japan

Authors and Affiliations

  1. Laboratory of Plant Cell Technology, Faculty of Horticulture, Chiba University, 648 Matsudo, Matsudo-City, Chiba, 271-8510, Japan

    T. Hirano, M. Mii & K. Ishikawa

  2. Botanic Gardens of Toyama, 42 Kamikutsuwada, Fuchu-machi, Nei-gun, Toyama, 939-2713, Japan

    T. Godo

Authors
  1. T. Hirano
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T. Godo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. Mii
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. K. Ishikawa
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to M. Mii.

Additional information

Communicated by K.K. Kamo

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hirano, T., Godo, T., Mii, M. et al. Cryopreservation of immature seeds of Bletilla striata by vitrification. Plant Cell Rep 23, 534–539 (2005). https://doi.org/10.1007/s00299-004-0893-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-004-0893-9

Keywords

Search

Navigation