Abstract
In the present study, the combination of the micronucleus test with analysis of the activity of the rRNA genes in mutagen-treated Hordeum vulgare (barley) by maleic hydrazide (MH) cells was performed. Simultaneously fluorescence in situ hybridization (FISH) with 25S rDNA as probes and an analysis of the transcriptional activity of 35S rRNA genes with silver staining were performed. The results showed that transcriptional activity is always maintained in the micronuclei although they are eliminated during the next cell cycle. The analysis of the transcriptional activity was extended to barley nuclei. MH influenced the fusion of the nucleoli in barley nuclei. The silver staining enabled detection of the nuclear bodies which arose after MH treatment. The results confirmed the usefulness of cytogenetic techniques in the characterization of micronuclei. Similar analyses can be now extended to other abiotic stresses to study the response of plant cells to the environment.
Highlights
Plant bioassays are commonly used to detect chromosome rearrangements induced by environmental, possibly mutagenic, factors
A micronucleus (MN) test combined with fluorescent in situ hybridization (FISH) allowed a quantitative analysis of the involvement of specific chromosome fragments in micronuclei formation and permitted the possible origin of mutagen-induced micronuclei to be explained. 35S rDNA are genes that belong to the category of “housekeeping genes”, which are present in every living cell and they have to have at least basal activity
H. vulgare is a very good model for such investigations due to its convenient karyotype features— large chromosomes, two of which display the presence of 35S rRNA genes, together with activity of all of them are convenient karyotype features that made barley a suitable experimental model for this study (Fig 1)
Summary
The combination of the micronucleus test with analysis of the activity of the rRNA genes in mutagen-treated Hordeum vulgare (barley) by maleic hydrazide (MH) cells was performed. Fluorescence in situ hybridization (FISH) with 25S rDNA as probes and an analysis of the transcriptional activity of 35S rRNA genes with silver staining were performed. The results showed that transcriptional activity is always maintained in the micronuclei they are eliminated during the cell cycle. The analysis of the transcriptional activity was extended to barley nuclei. MH influenced the fusion of the nucleoli in barley nuclei. The results confirmed the usefulness of cytogenetic techniques in the characterization of micronuclei. Similar analyses can be extended to other abiotic stresses to study the response of plant cells to the environment
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