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Three-Dimensional Acrylamide Fluorescence In Situ Hybridization for Plant Cells

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Plant Meiosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 990))

Abstract

Plant meiosis involves complex and dynamic processes that occur within the space inside the nucleus. Direct inspection of meiotic chromosomes by fluorescence microscopy has been used to investigate many of these processes. In particular, optical sectioning microscopy of fluorescence in situ hybridization (FISH)-stained nuclei provides three-dimensional spatial information about the organization and distribution of specific sequences and chromosomal loci within the nucleus. Here we provide a fully detailed three-dimensional (3D) acrylamide FISH method for the analysis of plant meiotic nuclei. Several examples illustrate the versatility of this technique for the investigation of meiotic telomere dynamics in maize, Arabidopsis, and oat. Additional examples of 3D FISH include chromosome painting in a maize chromosome-addition line of oat and telomere FISH with maize nuclei from plants expressing a fluorescently tagged fusion protein, histone H2B-mCherry.

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Acknowledgments

We would like to thank A.B. Thistle and D.L. Vera for critical reading of the manuscript and insightful comments and T.E. Clemente for providing the histone H2B-mCherry line of maize. This work was supported by a Women in Science Math and Engineering Fellowship to ESH, an American Heart Association predoctoral fellowship to SPM (AHA, Greater Southeast Affiliate, number 0715487B), a CRC-planning grant to HWB (2008), and a National Science Foundation grant to HWB (NSF IOS-1025954).

Author information

Authors and Affiliations

  1. Department of Biological Science, The Florida State University, Tallahassee, FL, USA

    Elizabeth S. Howe & Hank W. Bass

  2. Institute of Molecular Biophysics, The Florida State University, Tallahassee, FL, USA

    Shaun P. Murphy & Hank W. Bass

Authors
  1. Elizabeth S. Howe
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  2. Shaun P. Murphy
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  3. Hank W. Bass
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Editor information

Editors and Affiliations

  1. , Dept. of Plant Breeding & Genetics, Cornell University, Bradfield Hall 401, Ithaca, 14853, New York, USA

    Wojciech P. Pawlowski

  2. , Centre de Versailles-Grignon, Inst National de la Recherche Agronomiqu, Route de St-Cyr (RD10), Versailles, F-78000, France

    Mathilde Grelon

  3. School of Biosciences, The University of Birmingham, Edgbaston, Birmingham, B15 2TT, United Kingdom

    Susan Armstrong

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Howe, E.S., Murphy, S.P., Bass, H.W. (2013). Three-Dimensional Acrylamide Fluorescence In Situ Hybridization for Plant Cells. In: Pawlowski, W., Grelon, M., Armstrong, S. (eds) Plant Meiosis. Methods in Molecular Biology, vol 990. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-333-6_6

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  • DOI: https://doi.org/10.1007/978-1-62703-333-6_6

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-332-9

  • Online ISBN: 978-1-62703-333-6

  • eBook Packages: Springer Protocols

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