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Multiplication and cryopreservation of vanilla (Vanilla planifolia ‘Andrews’)

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Abstract

A simple and efficient method for multiplication of vanilla (Vanilla planifolia) was developed using in vitro fragmented explants (IFEs) as propagules. IFEs were obtained after dissecting apices from in vitro propagated clusters of plantlets, by cutting the remaining base of these plant clusters into segments of about 1 cm in length. After 4 months of culture on multiplication medium, 100% of IFEs produced up to 15 new shoots per explant, providing an efficient additional method for in vitro propagation of vanilla that maximizes the use of available material. Cryopreservation of apices from in vitro grown plants was achieved using the droplet vitrification protocol. Maximum survival (30%) and further regeneration (10%) of new shoots were obtained for apices derived from clusters of in vitro plantlets produced from microcuttings through a three-step droplet vitrification protocol: 1-d preculture of apices on solid MS medium with 0.3 M sucrose; loading with a 0.4 M sucrose + 2 M glycerol solution for 20–30 min; and exposure to plant vitrification solution PVS3 for 30 min at room temperature. Even though the cryogenic protocol needs to be optimized to improve results, this work represents the first successful report of cryopreservation of vanilla apices.

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Acknowledgments

Financial support of FOMIX project 37551 sponsored by CONACyT- Gobierno del Estado de Veracruz, Mexico; and CONACyT project of Complementary support are gratefully acknowledged by the first author.

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Authors and Affiliations

  1. Facultad de Ciencias Químicas, Universidad Veracruzana, Prolongación Oriente. 6, No. 1009, 94340, Orizaba, Veracruz, México

    Maria Teresa Gonzalez-Arnao, Claudia Esther Lazaro-Vallejo, Roberto Gamez-Pastrana, Yolanda Maria Martinez-Ocampo, Miriam Cristina Pastelin-Solano & Carlos Diaz-Ramos

  2. Institut de recherche pour le développement (IRD), UMR DIAPC, 911 avenue Agropolis, BP 64501, 34394, Montpellier Cedex 5, France

    Florent Engelmann

  3. Bioversity International, Via dei Tre Denari 472/a, 00057 Maccarese (Fiumicino), Rome, Italy

    Florent Engelmann

Authors
  1. Maria Teresa Gonzalez-Arnao
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  2. Claudia Esther Lazaro-Vallejo
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  3. Florent Engelmann
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  4. Roberto Gamez-Pastrana
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  5. Yolanda Maria Martinez-Ocampo
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  6. Miriam Cristina Pastelin-Solano
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  7. Carlos Diaz-Ramos
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Correspondence to Maria Teresa Gonzalez-Arnao.

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Editor: P. Druart

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Gonzalez-Arnao, M.T., Lazaro-Vallejo, C.E., Engelmann, F. et al. Multiplication and cryopreservation of vanilla (Vanilla planifolia ‘Andrews’). In Vitro Cell.Dev.Biol.-Plant 45, 574–582 (2009). https://doi.org/10.1007/s11627-009-9242-6

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  • DOI: https://doi.org/10.1007/s11627-009-9242-6

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