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Cryopreservation of in vitro-grown shoot tips of Clinopodium odorum using aluminium cryo-plates

  • Plant Tissue Culture
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Abstract

In this study, we applied the vitrification (V) cryo-plate protocol for cryopreservation of Clinopodium odorum in vitro-derived shoot tips. We studied the recovery of shoot tips after modification of various parameters of the V cryo-plate protocol, including sucrose concentration in the preconditioning medium, composition of the loading solution (LS) and vitrification solution (VS) and duration of treatment with PVS2 VS, which contained (w/v) 30% glycerol, 15% dimethylsulphoxide, 15% ethylene glycol and 13.7% sucrose. We also compared the efficiency of the V cryo-plate protocol with the dehydration (D) cryo-plate protocol. The optimal conditions determined for the V cryo-plate protocol included a 24-h preconditioning treatment on medium with 0.3 M sucrose; treatment for 20 min at room temperature with LS containing 2.0 M glycerol and 0.4 M sucrose; and dehydration with PVS2 for 60 min at 0°C. Under these conditions, 71.0% recovery of cryopreserved shoot tips was achieved. Only 29.2% regeneration was noted with the D cryo-plate protocol.

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References

  • Barboza GE, Cantero JJ, Nuñez PA, Ariza Espinar L (2009) Medicinal plants: review and a phytochemical and ethnopharmacological screening of the native Argentine flora, 34th edn. Univ Kurtziana, Nacional de Cordoba, Argentina

    Google Scholar 

  • Barraco G, Sylvestre I, Engelmann F (2011a) Cryopreservation of sugarcane (Saccharum spp.) shoot tips using encapsulation and droplet-vitrification. Sci Hortic 130:320–324

    Article  CAS  Google Scholar 

  • Barraco G, Sylvestre I, Iapichino G, Engelmann F (2011b) Cryopreservation of Limonium serotinum apical shoots from in vitro plantlets using droplet-vitrification. Sci Hortic 130:309–313

    Article  CAS  Google Scholar 

  • Bouman H, Tiekstra A, Petutschnig E, Homan M, Schreurs R (2003) Cryopreservation of Lilium species and cultivars. Acta Hortic 612:147–154

    Google Scholar 

  • Cantero JJ, Bianco CA (1986) Las Plantas Vasculares del Suroeste de la Provincia de Cordoba. Catalogo Preliminar de las Especies. Rev Univ Nacional De Rıo Cuarto 6:65–75 (in Spanish)

    Google Scholar 

  • Diaz MS, Palacio L, Figueroa AC, Goleniowski ME (2012) In vitro propagation of Muña-Muña (Clinopodium odorum (Griseb.) Harley). Biotech Res Intl. doi:10.1155/2012/196583

    Google Scholar 

  • Engelmann F (2011) Use of biotechnologies for the conservation of plant biodiversity. In Vitro Cell Dev Biol – Plant 47:5–16

    Article  Google Scholar 

  • Engelmann F (2014) Cryopreservation of clonal crops: a review of key parameters. Acta Hortic 1039:31–39

    Google Scholar 

  • Goleniowski ME, Bongiovanni GA, Palacio L, Nuñez CO, Cantero JJ (2006) Medicinal plants from the “Sierra de Comechingones”, Argentina. J Ethnopharmacol 107:324–341

    Article  PubMed  Google Scholar 

  • Harley RM, Paucar AG (2000) List of species of tropical American Clinopodium (Labiatae), with new combinations. Kew Bull 55:917–927

    Article  Google Scholar 

  • Kim HH, Lee YG, Ko HC, Park SU, Gwag JG, Cho EG, Engelmann F (2009a) Development of alternative loading solutions in droplet-vitrification procedures. CryoLetters 30:291–299

    CAS  PubMed  Google Scholar 

  • Kim HH, Lee YG, Shin DJ, Kim T, Cho EG, Engelmann F (2009b) Development of alternative plant vitrification solutions in droplet-vitrification procedures. CryoLetters 30:320–334

    CAS  PubMed  Google Scholar 

  • Mahady GB (2005) Medicinal plants for the prevention and treatment of bacterial infections. Curr Pharma Design 11:2405–2427

    Article  CAS  Google Scholar 

  • Mari S, Engelmann F, Chabrillange N, Huet C, Michaux-Ferrière N (1995) Histo-cytological study of coffee (Coffea racemosa and C. sessiliflora) apices of in vitro plantlets during their cryopreservation using the encapsulation-dehydration technique. CryoLetters 16:289–298

    Google Scholar 

  • Martinez GJ, Planchuelo AM, Fuentes E, Ojeda M (2006) A numeric index to establish conservation priorities for medicinal plants in the Paravachasca Valley, Cordoba, Argentina. Biodiv Conserv 15:2457–2475

    Article  Google Scholar 

  • 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 13:442–446

    Article  CAS  PubMed  Google Scholar 

  • Matsumoto T, Yamamoto S, Fukui K, Niino T (2013) Cryopreservation of persimmon shoot tips using D cryo-plate procedure. In Abstract: 2nd Intl. Symp. on Plant Cryopreservation. Fort Collins, USA, 11–14 August 2013, p 96

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Niino T, Yamamoto S, Fukui K, Castillo Martinez CR, Valle Arizaga M, Matsumoto T, Engelmann F (2013) Dehydration improves cryopreservation of mat rush (Juncus decipiens Nakai) basal stem buds on cryo-plates. CryoLetters 34:549–560

    CAS  PubMed  Google Scholar 

  • Niino T, Wunna WK, Nohara N, Rafique T, Yamamoto S, Fukui K, Valle Arizaga M, Castillo Martinez CR, Matsumoto T, Engelmann F (2014) Cryopreservation of mat rush lateral buds by air dehydration using aluminum cryo-plate. Plant Biotech Rep 31:281–287

    Article  CAS  Google Scholar 

  • 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 91:67–73

    Article  CAS  Google Scholar 

  • Panis B, Piette B, Swennen R (2005) Droplet vitrification of apical meristems: a cryopreservation protocol applicable to all Musaceae. Plant Sci 168:45–55

    Article  CAS  Google Scholar 

  • Panis B, Piette B, André E, Van den Houwe I, Swennen R (2011) Droplet vitrification: the first generic cryopreservation protocol for organized plant tissues? Acta Hortic 908:157–163

    Google Scholar 

  • R Foundation for Statistical Computing (2013) R: a Language and Environment for Statistical Computing. Version 3.0.1 (2013-05-16). http://www.r-project.org. Accessed 24 Oct 2014

  • Sakai A, Engelmann F (2007) Vitrification, encapsulation–vitrification and droplet–vitrification: a review. CryoLetters 28:151–172

    CAS  PubMed  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  CAS  PubMed  Google Scholar 

  • Salma M, Fki L, Engelmann-Sylvestre I, Niino T, Engelmann F (2014) Comparison of droplet-vitrification and D-cryoplate for cryopreservation of date palm (Phoenix dactylifera L.) polyembryonic masses. Sci Hortic 179:91–97

    Article  CAS  Google Scholar 

  • Sekizawa K, Yamamoto S, Rafique T, Fukui K, Niino T (2011) Cryopreservation of in vitro-grown shoot tips of carnation (Dianthus caryophyllus L.) by vitrification method using aluminium cryo-plates. Plant Biotech 28:401–405

    Article  Google Scholar 

  • Vujovic T, Sylvestre I, Ruzic D, Engelmann F (2011) Droplet-vitrification of apical shoot tips of Rubus fruticosus L. and Prunus cerasifera Ehrh. Sci Hortic 130:222–228

    Article  Google Scholar 

  • Yamamoto S, Rafique T, Priyantha WS, Fukui K, Matsumoto T, Niino T (2011) Development of a cryopreservation procedure using aluminium cryo-plates. CryoLetters 32:256–265

    CAS  PubMed  Google Scholar 

  • Yamamoto S, Fukui K, Rafique T, Khan NI, Castillo Martinez CR, Sekizawa K, Matsumoto T, Niino T (2012a) Cryopreservation of in vitro-grown shoot tips of strawberry by the vitrification method using aluminium cryo-plates. Plant Genet Res: Charact Util 10:14–19

    Article  CAS  Google Scholar 

  • Yamamoto S, Rafique T, Fukui K, Sekizawa K, Koyama A, Ichihashi T, Niino T (2012b) Development of an effective cryopreservation protocol using aluminium cryo-plates for in vitro-grown shoot tips of mulberries (Morus spp.) originated from the tropics and subtropics. Sanshi-Konchu Biotechnol 81:57–62 (in Japanese)

    Google Scholar 

  • Yamamoto S, Rafique T, Fukui K, Sekizawa K, Niino T (2012c) V-Cryo-plate procedure as an effective protocol for cryobanks. Case study of mint cryopreservation. CryoLetters 33:12–23

    CAS  PubMed  Google Scholar 

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Acknowledgements

This work has been performed in the framework of the AMCRYO project, funded by Agropolis Fondation (project no. 1202–007). Dr. Maria Ester Goleniowski (CEPROCOR, Cordoba, Argentina) introduced the plant material in vitro and performed the initial multiplication of cultures. We thank Lionel Feuillassier for his assistance in performing the statistical analysis of results.

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  1. IRD, UMR DIADE, 911 avenue Agropolis, 34394, Montpellier cedex 5, France

    Isabelle Engelmann-Sylvestre & Florent Engelmann

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  1. Isabelle Engelmann-Sylvestre
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  2. Florent Engelmann
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Correspondence to Florent Engelmann.

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Editor: John Forster

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Engelmann-Sylvestre, I., Engelmann, F. Cryopreservation of in vitro-grown shoot tips of Clinopodium odorum using aluminium cryo-plates. In Vitro Cell.Dev.Biol.-Plant 51, 185–191 (2015). https://doi.org/10.1007/s11627-015-9668-y

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  • DOI: https://doi.org/10.1007/s11627-015-9668-y

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