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Immunolocalization of AGPs and Pectins in Quercus suber Gametophytic Structures

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Plant Germline Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1669))

Abstract

The arabinogalactan proteins (AGPs) are highly glycosylated proteins, ubiquitous in plants that have been linked to numerous aspects of sexual reproduction in several plant species, including the monoecious tree species Quercus suber. AGPs are found in cell membranes and cell walls of all types of tissues, including reproductive cells and organs. Pectins are cell wall components that also have been shown to change in composition and quantity during the maturations of the male and female gametophyte in cork oak. These findings were only possible to reveal, due to the histological study of AGP and pectins epitopes by immunolabeling. The immunofluorescence microscopy technique uses antibodies linked to fluorophores and relies on the specificity of the antibody binding to its antigen, labeling the epitope with a fluorescent dye.

In the method presented here, we explore the immunolocalization technique performed in male and female flowers of Quercus suber, using London Resin (LR-White) as the embedding medium, after vacuum fixation with formaldehyde/glutaraldehyde. An extensive description of all the aspects of this technique is provided, from the plant material developmental stages selection to the critical analysis of results performed, continuously supported by troubleshooting recommendations.

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Authors and Affiliations

  1. Departamento de Biologia, Faculdade de Ciências, Universidade do Porto, Edifício FC4, Rua do Campo Alegre, 4169-007, Porto, Portugal

    Mário Luís da Costa, Ana Lúcia Lopes, Maria Isabel Amorim & Sílvia Coimbra

  2. Biosystems and Integrative Sciences Institute (BioISI), Porto, Portugal

    Mário Luís da Costa, Ana Lúcia Lopes, Maria Isabel Amorim & Sílvia Coimbra

Authors
  1. Mário Luís da Costa
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  2. Ana Lúcia Lopes
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  3. Maria Isabel Amorim
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  4. Sílvia Coimbra
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Corresponding author

Correspondence to Sílvia Coimbra .

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Editors and Affiliations

  1. Centre for Organismal Studies, University of Heidelberg, Heidelberg, Germany

    Anja Schmidt

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da Costa, M.L., Lopes, A.L., Amorim, M.I., Coimbra, S. (2017). Immunolocalization of AGPs and Pectins in Quercus suber Gametophytic Structures. In: Schmidt, A. (eds) Plant Germline Development. Methods in Molecular Biology, vol 1669. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7286-9_11

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  • DOI: https://doi.org/10.1007/978-1-4939-7286-9_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7285-2

  • Online ISBN: 978-1-4939-7286-9

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