Abstract
Alginate beads containing axillary buds of in vitro-grown gentian (Gentiana scabra Bunge var. buergeri Maxim.), were successfully cryopreserved following 2 step-preculture with sucrose and desiccation. The optimal preculture conditions were as follows: axillary buds were excised from in vitro-grown gentian plants and precultured on semi-solid Murashige and Skoog (MS) medium containing 0.1 M sucrose for 10 days (25 °C, 16-h photoperiod) (first step). This was followed by incubation on semi-solid MS media containing 0.4 M (1 day) and then 0.7 M sucrose (1 day) (second step). After preculture, the buds were encapsulated in alginate beads and desiccated aseptically on silica gel for 9 h to a water content of 10% (fresh weight basis), followed by immersion in liquid nitrogen (LN). With this protocol, 87% of the gentian buds survived exposure to LN and showed normal development of shoots and roots in vitro and in vivo. Depletion of NH4NO3 in the regeneration medium did not improve survival following desiccation and exposure to LN. The results show that 2 step-preculture with sucrose is effectively applicable in encapsulation–desiccation based cryopreservation of gentian axillary buds. This preculture can replace the conventionally used lengthy cold-hardening treatment and is useful for routine cryopreservation of gentian germplasm.
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Abbreviations
- LN – liquid nitrogen:
-
MS medium – Murashige and Skoog (1962) medium
References
MH Bhatti T Percival CDM Davet GG Henshaw D Blakesley (1997) ArticleTitleCryopreservation of embryogenic tissue of a range of genotypes of sweet potato (Ipomoea batatas [L] Lam.) using an encapsulation protocol Plant Cell Rep 16 802–806 Occurrence Handle10.1007/s002990050324
M Brison MT Boucaud Particlede F Dosba (1995) ArticleTitleCryopreservation of in vitro-grown shoot tips of two interspecific Prunus rootstocks Plant Sci 105 235–242 Occurrence Handle10.1016/0168-9452(94)04045-1
Y Chang BM Reed (2000) ArticleTitleExtended alternating-temperature cold acclimation and culture duration improve pear shoot cryopreservation Cryobiol 40 311–322 Occurrence Handle10.1006/cryo.2000.2251
J Dereuddre C Scottez Y Arnaud M Duron (1990) ArticleTitleResistance of alginate-coated axillary shoot tips of pear tree (Pyrus communis L. Beurré Hardy) in vitro plantlets to dehydration and subsequent freezing in liquid nitrogen: Effect of previous cold-hardening C. R. Acad. Sci. Paris ser. III 310 317–323
MT Gonzalez-Arnao J Juarez C Ortega L Navarro N Duran-Vila (2003) ArticleTitleCryopreservation of ovules and somatic embryos of Citrus using the encapsulation-dehydration technique Cryo-Lett 24 85–94
B Grout (1995) Genetic Preservation of Plant Cells in Vitro Springer Berlin 1–169
Kikuchi M & Yashiro N (1995) Cryopreservation of lateral buds of gentian by the encapsulation–dehydration technique. Japanese Society for Plant Tissue Culture, Program and Abstracts of the 14th Meeting (p. 74)
A Kuriyama K Watanabe S Ueno H Mitsuda (1990) ArticleTitleInhibitory effect of ammonium ion on recovery of cryopreserved rice cells Plant Sci 64 231–235 Occurrence Handle10.1016/0168-9452(89)90028-9
T Murashige F Skoog (1962) ArticleTitleA revised medium for rapid growth and bio assays with tobacco tissue cultures Physiol. Plant 15 473–497
T Niino A Sakai (1992) ArticleTitleCryopreservation of alginate-coated in vitro-grown shoot tips of apple, pear and mulberry Plant Sci 87 199–206 Occurrence Handle10.1016/0168-9452(92)90151-B
P Plessis C Leddet J Dereuddre (1991) ArticleTitleResistance to dehydration and freezing in liquid nitrogen of alginate-coated shoot tips of grape vine (Vitis vinifera L. cv. Chardonnay) C. R. Acad. Sci., Paris, ser. III 313 373–381
BM Reed HB Ragerstedt (1987) ArticleTitleFreeze preservation of apical meristems of Rubus in liquid nitrogen Hort. Sci 22 302–303
L Ryynanen (1998) ArticleTitleEffect of abscisic acid, cold hardening, and photoperiod on recovery of cryopreserved in vitro shoot tips of silver birch Cryobiol 36 32–39 Occurrence Handle10.1006/cryo.1997.2067 Occurrence Handle9500934
M Suzuki M Ishikawa T Akihama (1998) ArticleTitleA novel preculture method for induction of desiccation tolerance in gentian axillary buds for cryopreservation Plant Sci 135 69–76 Occurrence Handle10.1016/S0168-9452(98)00054-5
M Suzuki T Niino T Akihama (1994) ArticleTitleCryopreservation of shoot tips of kiwifruit seedlings by the alginate encapsulation-dehydration technique Plant Tiss. Cult. Lett 11 122–128
Suzuki M, Okuda H, Noda K, Ogiwara I, Shimura I, Akihama T & Ishikawa M (2005) Physiological changes in gentian axillary buds during 2 step-preculturing with sucrose that conferred high levels of tolerance to desiccation and cryopreservation. Ann. Bot. (accepted)
D Tanaka T Niino K Isuzugawa T Hikage M Uemura (2004) ArticleTitleCryopreservation of shoot apices of in vitro grown gentian plants: comparison of vitrification and encapsulation-vitrification protocols Cryo-Lett 25 167– 176
A Uragami MO Lucas J Ralambosoa M Renard J Dereuddre (1993) ArticleTitleCryopreservation of microspore embryos of oilseed rape (Brassica napus L.) by dehydration in air with or without alginate encapsulation Cryo-Lett 14 83–90
Y Wu F Engelmann Y Zhao M Zhou S Chen (1999) ArticleTitleCryopreservation of apple shoot tips: Importance of cryopreservation technique and of conditioning of donor plants Cryo-Lett 20 121–130
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Suzuki, M., Akihama, T. & Ishikawa, M. Cryopreservation of encapsulated gentian axillary buds following 2 step-preculture with sucrose and desiccation. Plant Cell Tiss Organ Cult 83, 115–121 (2005). https://doi.org/10.1007/s11240-005-4854-y
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DOI: https://doi.org/10.1007/s11240-005-4854-y