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Using Flow Cytometry Analysis in Plant Tissue Culture Derived Plants

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Plant Cell Culture Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1815))

Abstract

Somaclonal variation (SC) in plants regenerated from tissue culture, via organogenesis or somatic embryogenesis, is frequently associated with abnormalities in the content of deoxyribonucleic acid (DNA), viz., aneuploidy and polyploidy. Flow cytometry (FCM) using the nucleic acid-specific fluorochrome propidium iodide has proven to be a rapid, simple, and reproducible technique for assessment of DNA content and ploidy variation occurring in plant tissue cultures. Here an outline of the sample preparation of suspension with intact nuclei by the two-step standard method, and FCM analysis of DNA ploidy stability in plants regenerated from embryogenic cell suspension (ECS) of banana Musa acuminata, AAA, cv. Grand Naine (Cavendish subgroup) using an internal standard is described.

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Acknowledgments

The authors would like to express their gratitude to the government of México through SAGARPA-CONACYT Research Project No. 0048160 (RME) and studentship project (# 0048160) to MJBT.

Author information

Authors and Affiliations

  1. Unidad de Bioquímica y Biología Molecular de Plantas, Centro de Investigación Científica de Yucatán A.C., Mérida, Yucatán, Mexico

    Rosa María Escobedo-Gracia-Medrano, Martha Josefa Burgos-Tan & José Roberto Ku-Cauich

  2. Unidad de Biotecnología, Centro de Investigación Científica de Yucatán A.C., Mérida, Yucatán, Mexico

    Adriana Quiroz-Moreno

Authors
  1. Rosa María Escobedo-Gracia-Medrano
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  2. Martha Josefa Burgos-Tan
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  3. José Roberto Ku-Cauich
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  4. Adriana Quiroz-Moreno
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Corresponding author

Correspondence to Rosa María Escobedo-Gracia-Medrano .

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Editors and Affiliations

  1. Centro de Investigación Científica de Yucatán, Unidad de Bioquímica y Biología, Molecular de Plantas, Mérida, Yucatán, Mexico

    Víctor M. Loyola-Vargas

  2. Departamento de Ingeniería Genética, Unidad Irapuato, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Irapuato, Guanajuato, Mexico

    Neftalí Ochoa-Alejo

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Escobedo-Gracia-Medrano, R.M., Burgos-Tan, M.J., Ku-Cauich, J.R., Quiroz-Moreno, A. (2018). Using Flow Cytometry Analysis in Plant Tissue Culture Derived Plants. In: Loyola-Vargas, V., Ochoa-Alejo, N. (eds) Plant Cell Culture Protocols. Methods in Molecular Biology, vol 1815. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8594-4_22

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  • DOI: https://doi.org/10.1007/978-1-4939-8594-4_22

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