Rapid detection of aneuploidy in Musa using flow cytometry (original) (raw)

Abstract

We report a procedure for the rapid and convenient detection of aneuploidy in triploid Musa using DNA flow cytometry. From a population of plants derived from gamma-irradiated shoot tips, plants were selected based on aberrant morphology and their chromosome numbers were counted. Aneuploids plants with chromosome numbers 2n=31 or 32 were found as well as the expected triploid plants (2n=3x=33). At the same time, the nuclear DNA content of all plants was measured using flow cytometry. The flow cytometric assay involved the use of nuclei isolated from chicken red blood cells (CRBC), which served as an internal reference standard. The relative DNA content of individual plants was expressed as a ratio of DNA content of CRBC and Musa (DNA index). In order to estimate the chromosome number using flow cytometry, the relative DNA content of plants with unknown ploidy was expressed as a percentage of the DNA content of triploid plants. The classification based on flow cytometry fully agreed with the results obtained by chromosome counting. The results indicated that flow cytometry is a convenient and rapid method for the detection of aneuploidy in Musa.

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Acknowledgements.

We thank Ir. I. van den Houwe, Musa curator of the INIBAP Transit Centre at the Katholieke Universiteit, Leuven, Belgium, for supplying the plant material. We are grateful to Mr. Chris Rigney and Mr. Mohan Jain for their critical reading of the manuscript. This study was undertaken as a part of the Global Programme for Musa Improvement (PROMUSA) and was supported by the International Atomic Energy Agency and the Directorate General for International Cooperation (DGIC), Belgium.

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Authors and Affiliations

1. Plant Breeding Unit, FAO/IAEA Agriculture and Biotechnology Laboratory, International Atomic Energy Agency Laboratories, 2444, Seibersdorf, Austria
   N. Roux, A. Toloza & F. J. Zapata-Arias
2. Chemistry Unit, Physics-Chemistry-Instrumentation Laboratory, International Atomic Energy Agency Laboratories, 2444 , Seibersdorf, Austria
   Z. Radecki
3. Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovska 6, 77200 , Olomouc, Czech Republic
   J. Dolezel

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1. N. Roux
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2. A. Toloza
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3. Z. Radecki
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4. F. J. Zapata-Arias
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5. J. Dolezel
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Correspondence toN. Roux.

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Communicated by A. Altman

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Roux, N., Toloza, A., Radecki, Z. et al. Rapid detection of aneuploidy in Musa using flow cytometry.Plant Cell Rep 21, 483–490 (2003). https://doi.org/10.1007/s00299-002-0512-6

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• Received: 25 January 2002
• Revised: 01 August 2002
• Accepted: 28 October 2002
• Published: 10 December 2002
• Issue Date: January 2003
• DOI: https://doi.org/10.1007/s00299-002-0512-6

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