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The chromosomes of Brassica species are small and poorly differentiated, and their identification is extremely difficult using conventional cytogenetic methods. Progress in molecular analysis of Brassica species requires cytogenetic maps of their chromosomes. Chromosome-specific markers are needed to distinguish particular pairs of homologous chromosomes and for karyotyping. In this study, three morphological groups of chromosomes in B. campestris (genome A) and B. oleracea (genome C) and two in B. nigra (genome B) were distinguished by the morphometric features of the chromosomes, based upon arm ratio and absolute length. Using fluorescence in situ hybridization and differential stainings it was possible to establish further markers for five pairs of chromosomes in genome A, three in genome B and two in genome C, and to present idiograms of the chromosomes for three diploid Brassica species. However, for clear identification of all chromosome pairs more cytogenetic markers are needed.
Different explants from diploid and tetraploid plants of Arabidopsis thaliana ecotype Wilna were cultured in vitro on the same medium in the same conditions. The mode and capacity for regeneration and the ploidy level of regenerants were examined in eight callus lines. The explant responses to in vitro conditions depended on the ploidy level of donor plants and on the type of explant. Callus derived from leaves of tetraploid plants showed the highest ability to regenerate. Histological investigation showed that plants regenerated mostly through organogenesis and occasionally through embryogenesis. Somatic embryos were observed more frequently in callus derived from tetraploid plants. Regenerated plants were diploids, triploids and tetraploids. The majority of regenerated plants from callus of diploid origin were diploid, but diploids were also observed quite frequently among regenerants from callus of tetraploid origin.
The present study is a rare example of a detailed characterization of chromosomal aberrations by identification of individual chromosomes (or chromosome arms) involved in their formation in plant cells by using fluorescent in situ hybridization (FISH). In addition, the first application of more than 2 DNA probes in FISH experiments in order to analyse chromosomal aberrations in plant cells is presented. Simultaneous FISH with 5S and 25 S rDNA and, after reprobing of preparations, telomeric and centromeric DNA sequences as probes, were used to compare the cytogenetic effects of 2 chemical mutagens: N-nitroso-N-methylurea (MNU) and maleic hydrazide (MH) on root tip meristem cells of Hordeum vulgare (2n =14). The micronucleus (MN) test combined with FISH allowed the quantitative analysis of the involvement of specific chromosome fragments in micronuclei formation and thus enabled the possible origin of mutagen-induced micronuclei to be explained. Terminal deletions were most frequently caused by MH and MNU. The analysis of the frequency of micronuclei with signals of the investigated DNA probes showed differences between the frequency of MH- and MNU-induced micronuclei with specific signals. The micronuclei with 2 signals, telomeric DNA and rDNA (5S and/or 25S rDNA), were the most frequently observed in the case of both mutagens, but with a higher frequency after treatment with MH (46%) than MNU (37%). Also, 10% of MH-induced micronuclei were characterized by the presence of only telomere DNA sequences, whereas there were almost 3-fold more in the case of MNU-induced micronuclei (28%). Additionally, by using FISH with the same probes, an attempt was made to identify the origin of chromosome fragments in mitotic anaphase.
In this study, a novel repetitive sequence pTaq10 was isolated from the Taq I digest of the genomic DNA of the pseudocereal Chenopodium quinoa. Sequence analysis indicated that this 286-bp monomer is not homologous to any known retroelement sequence. FISH and Southern blot analysis showed that this sequence is characterized by an interspersed genomic organization. After FISH, hybridization signals were observed as small dots spread throughout all of the chromosomes. pTaq hybridization signals were excluded from 45S rRNA gene loci, but they partly overlapped with 5S rDNA loci. pTaq10 is not a species-specific sequence, as it was also detected in C. berlandieri.
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