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Effect of antioxidants on the genetic stability of cryopreserved mint shoot tips by encapsulation–dehydration

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Abstract

The effect of antioxidants applied in one step of a cryopreservation protocol by encapsulation–dehydration on recovery and genetic stability of mint shoot tips has been studied. Glutathione (0.16 or 0.24 mM), ascorbic acid (0.28 or 0.43 mM) and α-tocopherol (vitamin E) were added to the preculture medium (0.3 M sucrose). DNA was extracted from three different types of samples: leaves from shoots, callus at the base of shoots and callus. RAPD and AFLP markers were used to assess the genetic stability. The use of antioxidants did not improve recovery after cryopreservation. One of the genotypes, ‘MEN 198’, showed higher percentage of stable samples than the other one, ‘MEN 186’ (56 vs. 37 %; considering all treatments and types of explant). The use of vitamin E improved the percentage of stable samples with respect to control treatment (no antioxidant) in ‘MEN 186’. No differences in the percentages of stable samples were observed among cryopreserved and non-cryopreserved (treated similarly without immersion in liquid nitrogen) plant material. Recovered shoots of both genotypes showed higher stability (76–80 % stable samples) than callus samples (14–22 %).

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Acknowledgments

This research was supported by the Spanish Government project AGL2010-21989-C02-01. C.K. is supported by a grant from Universidad Politécnica de Madrid. Technical support from C. Sansegundo, M. Huertas and C. Ruiz is appreciated.

Author Contributions

MEG-B: experiment design with special emphasis in the cryopreservation studies, manuscript writing. CK: performance of cryopreservation experiments and part of genetic analysis, partially manuscript writing. MAI: statistical analysis. CM: experiment design with special emphasis in the genetic studies, part of the genetic analysis, manuscript writing.

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Authors and Affiliations

  1. Departamento de Biotecnología-Biología Vegetal, Escuela Técnica Superior de Ingeniería Agronómica, Alimentaria y de Biosistemas, Universidad Politécnica de Madrid, Ciudad Universitaria, 28040, Madrid, Spain

    María Elena González-Benito, Carolina Kremer & Carmen Martín

  2. Dpto. Economía Agraria, Estadística y Gestión de Empresas, Escuela Técnica Superior de Ingeniería Agronómica, Alimentaria y de Biosistemas, Universidad Politécnica de Madrid, Ciudad Universitaria, 28040, Madrid, Spain

    Miguel A. Ibáñez

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  1. María Elena González-Benito
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  2. Carolina Kremer
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  3. Miguel A. Ibáñez
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  4. Carmen Martín
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Correspondence to María Elena González-Benito.

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González-Benito, M.E., Kremer, C., Ibáñez, M.A. et al. Effect of antioxidants on the genetic stability of cryopreserved mint shoot tips by encapsulation–dehydration. Plant Cell Tiss Organ Cult 127, 359–368 (2016). https://doi.org/10.1007/s11240-016-1056-8

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  • DOI: https://doi.org/10.1007/s11240-016-1056-8

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