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Improved somatic embryogenesis of grapevine (Vitis vinifera) with focus on induction parameters and efficient plant regeneration

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Abstract

A study of four parameters (induction medium, floral explant, developmental stage and year) was carried out to determine the best combination for the embryogenesis induction of eight grapevine (Vitis vinifera L.) cultivars. Anthers and ovaries were extracted from flower buds at three developmental phases and incubated in two induction media over two consecutive years. As average, the percentage of embryogenesis on Nitsch and Nitsch-derived medium (9.1%) was higher than in Murashige and Skoog-derived medium (5.9%) and embryogenesis from ovaries (10.1%) was 2-fold higher than from anthers (4.9%). Earlier flower developmental stages (II–III) favored embryogenic induction from anthers, while later stages (III–V) did it from ovaries. Induction of embryogenic cultures was genotype dependent. Two years after the establishment of the embryogenic lines, an average of 48.0% of the pro-embryogenic masses were viable and suitable to initiate cell suspensions. Embryogenic cultures of four genotypes showed a high percentage of conversion from embryos to plants: Albariño (61.8%), Garnacha (48.8%), Tempanillo (71.0%) and Sultanina (69.0%). Moreover, cell suspensions were competent for transient transformation based on β-glucuronidase assay, as up to 6,387 blue spots per Petri plate after Biolistic bombardment were obtained. Here, we present the advantage of ovaries over anthers for the embryogenesis induction of several grapevine cultivars. This is the first report of embryogenesis from the cultivars Albariño, Verdejo and Muscat Hamburg as well as transient transformation of Albariño and Tempranillo.

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Abbreviations

2, 4-D:

2,4-Dichlorophenoxyacetic acid

4-CPPU:

N-(2-chloro-4-pyridyl)-N′-phenylurea

BAP:

6-Benzylaminopurine

NOA:

β-Naphthoxyacetic acid

MS:

Murashige and Skoog medium

NN:

Nitsch and Nitsch medium

PEM:

Pro-embryogenic masses

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Acknowledgments

We gratefully acknowledge financial support from a Marie Curie International Re-integration Grant (MIRG-2005-28341) of the European Commission and an I + D + i Grant (AGL-2006-5856) of the Spanish Ministry of Education and Science (MEC). JR Vidal was fully supported by a Ramón y Cajal contract from MEC. Thanks are also extended to F. Cabello for allowing us to collect flower buds from vineyards at IMIDRA (Comunidad de Madrid) and to MJ Carmona for the use of her laboratory facilities.

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Authors and Affiliations

  1. Department of Plant Physiology, Universidad de Santiago de Compostela, Campus Sur, 15782, Santiago de Compostela, Spain

    J. R. Vidal, J. Rama, L. Taboada & A. Segura

  2. Department of Biotechnology, Universidad Politécnica de Madrid, Ciudad Universitaria, 28040, Madrid, Spain

    J. R. Vidal

  3. Department of Plant Biology, Universidad Politécnica de Madrid, Ciudad Universitaria, 28040, Madrid, Spain

    C. Martin & M. E. González-Benito

  4. Department of Statistics, Universidad Politécnica de Madrid, Ciudad Universitaria, 28040, Madrid, Spain

    M. Ibañez

Authors
  1. J. R. Vidal
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  2. J. Rama
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  3. L. Taboada
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  4. C. Martin
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  6. A. Segura
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  7. M. E. González-Benito
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Correspondence to J. R. Vidal.

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Vidal, J.R., Rama, J., Taboada, L. et al. Improved somatic embryogenesis of grapevine (Vitis vinifera) with focus on induction parameters and efficient plant regeneration. Plant Cell Tiss Organ Cult 96, 85–94 (2009). https://doi.org/10.1007/s11240-008-9464-z

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  • DOI: https://doi.org/10.1007/s11240-008-9464-z

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