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Plant Meiosis pp 117-129 | Cite as

Cytological Techniques to Study Cytomixis in Plant Male Meiosis

  • Sergey R. Mursalimov
  • Yuriy V. Sidorchuk
  • Elena V. Deineko
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2061)

Abstract

In this chapter we describe cytological techniques to study cytomixis, a process of nuclear migration between plant cells, in squashed plant male meiocytes of Nicotiana tabacum and Secale cereale. To perform immunostaining or fluorescence in situ hybridization (FISH) on meiotic cells involved in cytomixis common protocols are modified. During preparation of specimens for subsequent cytological analysis, it is necessary not only to make DNA and proteins accessible to DNA probes and antibodies, but also to preserve cell cytoplasm. There are also some important modifications in the protocols applied for meiocytes of different plant species. Here we describe protocols for immunostaining and FISH in rigid tobacco male meiocytes with dense cytoplasm and thick callose wall, that tolerate hard squashing, and in soft rye male meiocytes, that are easily damaged upon squashing, both to study cytomixis.

Key words

Cytomixis Tobacco Rye Meiosis Immunolocalization 

Notes

Acknowledgments

Rye seeds were kindly provided by Silkova O.G. (Institute of Cytology and Genetics, Russia). The work was supported by the Russian Foundation for Basic Research [16-34-60007 mol_a_dk] and Siberian Branch of the Russian Academy of Science under the program “Molecular genetic bases of regulation of genes expression, morphology, differentiation and cell reprogramming” [0324-2019-0042].

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Sergey R. Mursalimov
    • 1
  • Yuriy V. Sidorchuk
    • 1
  • Elena V. Deineko
    • 1
  1. 1.Institute of Cytology and GeneticsRussian Academy of Sciences, Siberian BranchNovosibirskRussian Federation

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