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Evaluation of Analytical Techniques to Predict Viability after Cryopreservation

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Abstract

Preservation of plant germplasm is important to safeguard biodiversity and to store elite plants. Cryopreservation is one of the possible preservation techniques. Research for a cryopreservation protocol is often inefficient because of slow or poor regrowth of plant material. Therefore, at least one technique, that allows a quick and accurate prognosis of viability after cryopreservation, is required. We evaluated five techniques: electrolyte leakage, triphe-nyltetrazoliumchloride (TTC) staining (visual and spectrophotometrical analysis), malondialdehyde concentrations in plant tissue and a mathematical model that relates ‘water content’ to the weight of encapsulated plant material. Electrolyte leakage and TTC-staining (if visually analysed) are efficient to predict viability. Our mathematical model allows us to save time and plant material in order to develop an efficient encapsulation—dehydration protocol. All other techniques were rejected because of the high variability of the results. This is due to the variability of biochemical activity in plant tissue and the small amount of tissue used in the experiments.

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References

  • Benson EE & Withers LA (1987) Gas chromatographic analysis of volatile hydrocarbon production by cryopreserved plant tissue cultures: a non destructive method for assessing stability. Cryo Lett. 8: 35–46

    CAS  Google Scholar 

  • Bouman H & De Klerk GJ (1990) Cryopreservation of lily meristems. Acta Hortic. 266: 331–337

    Google Scholar 

  • Brison M, De Boucaud MT & Dosba F (1995) Cryopreservation of in vitro grown shoot tips of two interspecific Prunus rootstocks. Plant Sci. 105: 235–242

    Article  CAS  Google Scholar 

  • Chevrier N, Sarhan F & Chung YS (1988) Oxydative damage and repair in Euglena gracilis exposed to ozone I SH groups and lipids. Plant Cell Physiol. 29: 321–327

    CAS  Google Scholar 

  • De Proft MP, Maene LJ & Debergh PC (1985) Carbon dioxide and ethylene evolution in the culture atmosphere of Magnolia cultured in vitro. Physiol. Plant. 65: 375–379

    CAS  Google Scholar 

  • Fukai S, Goi M & Tanaka M (1994) The chimeric structure of the apical dome of chrysanthemum (Dendranthema grandiflorum (Ramat.) Kitam.) is affected by cryopreservation. Sci. Hort. 57: 347–351

    Google Scholar 

  • Gonzalez-Arnao MT, Engelmann F, Villavicencio CU, Morenza M & Rios A (2000) Cryopreservation of citrus apices using the encapsulation-dehydration technique. In: Engelmann F & Takagi H (eds) Cryopreservation of Tropical Germplasm. Current Research Progress and Application (pp. 217–221). JIRCAS, Rome

    Google Scholar 

  • Grout BWW (1991) Cryopreservation of plants cells and organs. In: Zakri AH, Normah MT, Senawi MT & Abdul Karim AG (eds) Conservation of Plant Genetic Resources Through in vitro Methods (pp. 43–45). FRIM/MNCPGR, Kuala Lumpur

    Google Scholar 

  • Grout BWW (1995a) Cryopreservation of excised meristems, shoot tips and somatic embryos. In: Grout BWW (ed) Genetic Preservation of Plant Cells in vitro (pp. 47–61). Springer, Berlin

    Google Scholar 

  • Grout BWW (1995b) Introduction to the in vitro preservation of plant cells, tissues and organs. In: Grout BWW (ed) Genetic Preservation of Plant Cells in vitro (pp. 1–20). Springer, Berlin

    Google Scholar 

  • Harding K & Benson EE (1995) Biochemical and molecular methods for assessing damage, recovery and stability in cryopreserved plant germplasm. In: Grout BWW (ed) Genetic Preservation of Plant Cells in vitro (pp. 113–167). Springer, Berlin

    Google Scholar 

  • Haskins RH & Kartha KK (1980) Freeze preservation of pea meristems: cell survival. Can. J. Bot. 58: 833–840

    Google Scholar 

  • Heath RL & Packer L (1968) Photoperoxydation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxydation. Arch. Biochem. Biophys. 125: 189–198

    CAS  PubMed  Google Scholar 

  • Jitsuyama Y, Suzuki T, Harada T & Fujikawa S (2002) Sucrose incubation increases freezing tolerance of asparagus (Asparagus officinalis L.) embryonic cell suspension. Cryo Lett. 23: 103–112

    CAS  Google Scholar 

  • Jouve L, Engelmann F, Noirot M & Charrier A (1993) Evaluation of biochemical markers (sugar, proline, malondialdehyde and ethylene) for cold sensitivity in microcuttings of two coffee species. Plant Sci. 91: 109–116

    Article  CAS  Google Scholar 

  • Kartha KK, Leung NL & Gamborg OL (1979) Freeze-preservation of pea meristems in liquid nitrogen and subsequent plant regeneration. Plant Sci. Lett. 15: 7–15

    Google Scholar 

  • Ketchie DO, Beeman CH & Ballard AL (1972) Relationship of electrolytic conductance to cold injury and acclimation in fruit trees. J. Am. Soc. Hort. Sci. 97: 403–406

    Google Scholar 

  • Lloyd G & McCown B (1980) Use of microculture for production and improvement of Rhododendron spp. HortScience 15: 416

    Google Scholar 

  • Lunec J & Dormandy TL (1979) Fluorescent lipid-peroxydation products in synovial fluid. Clin. Sci. 56: 53–59

    CAS  PubMed  Google Scholar 

  • Mandal BB (2000) Cryopreservation of yam apices: a comparative study with three different techniques. In: Engelmann F & Takagi H (eds) Cryopreservation of Tropical Germplasm. Current Research Progress and Application (pp. 233–237). JIRCAS, Rome

    Google Scholar 

  • Matsumoto T, Mochida K, Itamura H & Sakai A (2001) Cryopreservation of persimmon (Diospyros kaki Thunb.) by vitrification of dormant shoot tips. Plant Cell Rep. 20: 398–402

    CAS  Google Scholar 

  • Niino T & Sakai A (1992). Cryopreservation of alginate-coated in vitro-grown shoot tips of apple, pear and mulberry. Plant Sci. 87: 199–206

    Article  CAS  Google Scholar 

  • Niino T, Tahiro 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. Sci. Hortic. 70: 155–163

    Article  Google Scholar 

  • Ögren E, Nilsson T & Sundblad LG (1997) Relationship between respiratory depletion of sugars and loss of cold hardiness in coniferous seedlings over-wintering at raised temperatures: indications of different sensitivities of spruce and pine. Plant Cell Environ. 20: 247–253

    Google Scholar 

  • Paques M, Poisonnier M, Dumas E & Monod V (1997) Cryopreservation of dormant and non dormant broad-leaved trees. Acta Hortic. 447: 491–497

    Google Scholar 

  • Pellett NE & Heleba DA (1998) Comparing callus growth with discoloration and electrical conductivity as measures of stem injury after freezing woody plants. J. Am. Soc. Hort. Sci. 123: 826–831

    Google Scholar 

  • Sakai A, Kobayashi S & Oiyama I (1991) Survival by virification of nucellar cells of navel orange (Citrus sinensis var. Brasiliensis Tanaka) cooled to-196ºC. Plant Physiol. 9: 30–33

    Google Scholar 

  • Steponkus PL (1984) Role of the plasma membrane in freezing injury and cold acclimation. Annu. Rev. Plant Physiol. 35: 543–584

    Article  CAS  Google Scholar 

  • Steponkus PL & Lamphear FO (1967) Refinement of the triphenyl tetrazolium chloride method of determining cold injury. Plant Physiol. 42: 1423–1426

    CAS  Google Scholar 

  • Stuart NW (1939) Comparative cold hardiness of scion roots from fifty apple varieties. Proc. Am. Soc. Hort. Sci. 37: 330–334

    Google Scholar 

  • Takagi H (2000) Recent developments in cryopreservation of shoot apices of tropical species. In: Engelmann F & Takagi H (eds) Cryopreservation of Tropical Germplasm. Current Research Progress and Application (pp. 178–193). JIRCAS, Rome

    Google Scholar 

  • Touchell DH, Dixon KW & Tan B (1992) Cryopreservation of shoot-tips of Grevillea scapigera (Proteaceae): a rare and endangered plant from western Australia. Aust. J. Bot. 40: 305–310

    Article  Google Scholar 

  • Towill LE (1995) Cryopreservation by vitrification. In: Grout BWW (ed) Genetic Preservation of Plant Cells in vitro (pp. 99–111). Springer, Berlin

    Google Scholar 

  • Van Waes JM & Debergh PC (1986) In vitro germination of some West-European orchids. Physiol. Plant. 67: 253–261

    CAS  Google Scholar 

  • Wang QC, Batuman Ö, Li P, Bar-Joseph M & Gafny R (2002) Cryopreservation of in vitro-grown shoot tips of ‘Troyer’ citrange [poncirus trifoliate (L.) Raf. × Citrus sinensis (L.) Osbeck.] by encapsulation-dehydratation. Plant. Cell Rep. 20: 901–906

    CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Plant Production, Faculty of Agricultural and Applied Biological Sciences, Gent, Coupure Links 653, 9000, Gent, Belgium

    H. Verleysen, E. Van Bockstaele & P. Debergh

  2. Department of Plant Genetics and Breeding, CLO Gent, Caritasstraat 21, 9090, Melle, Belgium

    H. Verleysen, G. Samyn & E. Van Bockstaele

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  1. H. Verleysen
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  2. G. Samyn
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  3. E. Van Bockstaele
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  4. P. Debergh
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Correspondence to H. Verleysen.

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Verleysen, H., Samyn, G., Van Bockstaele, E. et al. Evaluation of Analytical Techniques to Predict Viability after Cryopreservation. Plant Cell, Tissue and Organ Culture 77, 11–21 (2004). https://doi.org/10.1023/B:TICU.0000016483.00158.a9

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  • DOI: https://doi.org/10.1023/B:TICU.0000016483.00158.a9

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