Comet-FISH for the evaluation of plant DNA damage after mutagenic treatmentsReport as inadecuate




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Journal of Applied Genetics

, Volume 54, Issue 4, pp 407–415

First Online: 01 October 2013Received: 20 May 2013Revised: 28 August 2013Accepted: 29 August 2013DOI: 10.1007-s13353-013-0169-6

Cite this article as: Kwasniewska, J. & Kwasniewski, M. J Appl Genetics 2013 54: 407. doi:10.1007-s13353-013-0169-6

Abstract

The aim of this study was to perform a comparative investigation of the actions of three mutagens that are widely used in plant mutagenesis using the comet-FISH technique. The comet-FISH technique was used for the analysis of DNA damage and the kinetics of repair within specific DNA sequences. FISH with rDNA and telomeric-centromeric DNA probes was applied to comets that were obtained from an alkaline-neutral comet assay. Migration within specific DNA sequences was analysed after treatment with two chemical mutagens-maleic hydrazide MH and N-nitroso-N-methylurea MNU, and γ-rays. Barley was used as a model plant in this study. The possible utility of specific DNA sequences in a comparative assessment of the distribution of DNA damage within a plant genome was evaluated. This study proved that the comet-FISH technique is suitable for a detailed quantification of DNA damage and repair within specific DNA sequences in plant mutagenesis. The analysis of FISH signals demonstrated that the involvement of specific DNA sequences in DNA damage was different and was dependent on the mutagen used. We showed that 5S rDNA and telomeric DNA sequences are more sensitive to mutagenic treatment, which was expressed by a stronger fragmentation and migration in comparison to the other probes used in the study. We found that 5S rDNA and telomeric DNA probes are more suitable for testing the genotoxicity of environmental factors. A comparison of the involvement of specific chromosome domains in direct DNA breakage-repair and in chromosome aberration formation after mutagen treatment indicates the compatibility of the results.

KeywordsBarley Centromere DNA fragmentation rDNA Telomere  Download fulltext PDF



Author: Jolanta Kwasniewska - Miroslaw Kwasniewski

Source: https://link.springer.com/







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