Molecular cytogenetic characterization of parental genomes in the partial amphidiploid Triticum aestivum x Thinopyrum ponticum

The wheat line PF 839197 and six hybrid derivatives from a cross between PF 839197 and Thinopyrum ponticum were cytologically characterized by fluorescent in situ hybridization (FISH). Probes for the 5S and 45S rDNA genes (pTa794 and pTa71, respectively), a highly repetitive rye sequence (pSc119.2), the synthetic oligonucleotide (AAG)5, and total genomic DNA from Th. ponticum and rye were used. In the wheat line, a 1RS.1BL translocation was revealed by the labeling patterns produced with pSc119.2 and (AAG)5, and confirmed by genomic in situ hybridization (GISH) using rye genomic DNA as a probe. Analyses of partial amphiploids confirmed previous results indicating mitotic instability, with a tendency to stabilize at 2n = 42 or 56. GISH with Th. ponticum genomic DNA showed that in one hybrid derivative, with lower chromosome numbers (2n = 42-45), chromosomes were not labeled, whereas in the hybrids with 2n = 48-56 up to 14 chromosomes were labeled. These data suggest that the original chromosome set of these hybrids was 2n = 56, and that chromosomes from both genomes were lost by mitotic instability. FISH using the rDNA probes and GISH with Thinopyrum genomic DNA suggested that cells with 2n = 56 contained an entire wheat genome plus two monoploid chromosome sets of Th. ponticum

Karyotype heterogeneity in Philodendron s.l. (Araceae) revealed by chromosome mapping of rDNA loci.

Philodendron s.l. (Araceae) has been recently focus of taxonomic and phylogenetic studies, but karyotypic data are limited to chromosome numbers and a few published genome sizes. In this work, karyotypes of 34 species of Philodendron s.l. (29 species of Philodendron and five of Thaumatophyllum), ranging from 2n = 28 to 36 chromosomes, were analyzed by fluorescence in situ hybridization (FISH) with rDNA and telomeric probes, aiming to understand the evolution of the karyotype diversity of the group. Philodendron presented a high number variation of 35S rDNA, ranging from two to 16 sites, which were mostly in the terminal region of the short arms, with nine species presenting heteromorphisms. In the case of Thaumatophyllum species, we observed a considerably lower variation, which ranged from two to four terminal sites. The distribution of the 5S rDNA clusters was more conserved, with two sites for most species, being preferably located interstitially in the long chromosome arms. For the telomeric probe, while exclusively terminal sites were observed for P. giganteum (2n = 30) chromosomes, P. callosum (2n = 28) presented an interstitial distribution associated with satellite DNA. rDNA sites of the analyzed species of Philodendron s.l. species were randomly distributed considering the phylogenetic context, probably due to rapid evolution and great diversity of these genomes. The observed heteromorphisms suggest the accumulation of repetitive DNA in the genomes of some species and the occurrence of chromosomal rearrangements along the karyotype evolution of the group

Chromosome characterization in Thinopyrum ponticum (Triticeae, Poaceae) using in situ hybridization with different DNA sequences

Thinopyrum ponticum (2n = 10x = 70, JJJJsJs) belongs to the Triticeae tribe, and is currently used as a source of pathogen resistance genes in wheat breeding. In order to characterize its chromosomes, the number and position of 45S and 5S rDNA sites, as well as the distribution of the repetitive DNA sequences pAs1 and pSc119.2, were identified by fluorescent in situ hybridization. The number of nucleoli and NORs was also recorded after silver nitrate staining. Seventeen 45S and twenty 5S rDNA sites were observed on the short arms of 17 chromosomes, the 45S rDNA was always located terminally. On three other chromosomes, only the 5S rDNA site was observed. Silver staining revealed a high number of Ag-NORs (14 to 17) on metaphase chromosomes, whereas on interphase nuclei there was a large variation in number of nucleoli (one to 15), most of them (82.8%) ranging between four and nine. The pAs1 probe hybridized to the terminal region of both arms of all 70 chromosomes. In addition, a disperse labeling was observed throughout the chromosomes, except in centromeric and most pericentromeric regions. When the pSc119.2 sequence was used as a probe, terminal labeling was observed on the short arms of 17 chromosomes and on the long arms of five others. The relative position of 45S and 5S rDNA sites, together with the hybridization pattern of pAs1 and pSc119.2 probes, should allow whole chromosomes or chromosome segments of Th. ponticum to be identified in inbred lines of wheat x Th. ponticum

Karyotype and genome size comparative analyses among six species of the oilseed-bearing genus Jatropha (Euphorbiaceae)

Abstract Jatropha is an important genus of Euphorbiaceae, with species largely used for various purposes, including the manufacturing of soaps and pharmaceutical products and applications in the bioenergetic industry. Although there have been several studies focusing J. curcas in various aspects, the karyotype features of Jatropha species are poorly known. Therefore, we analyzed six Jatropha species through fluorochrome staining (CMA/DAPI), fluorescent in situ hybridization (FISH) with 5S and 45S rDNA probes and genome size estimation by flow cytometry. Our results revealed several chromosome markers by both CMA/DAPI and FISH for the analyzed species. Five Jatropha species (J. curcas, J. gossypiifolia, J. integerrima, J. multifida and J. podagrica) showed four CMA-positive (CMA+) bands associated with the 5S and 45S rDNA sites (one and two pairs, respectively). However, J. mollissima displayed six CMA+/DAPI- bands co-localized with both 5S and 45S rDNA, which showed a FISH superposition. A gradual variation in the genome sizes was observed (2C = 0.64 to 0.86 pg), although an association between evidenced heterochromatin and genome sizes was not found among species. Except for the unique banding pattern of J. mollissima and the pericentromeric heterochromatin of J. curcas and J. podagrica, our data evidenced relatively conserved karyotypes

Genotoxic potential of the latex from cotton-leaf physicnut (Jatropha gossypiifolia L.)

Jatropha gossypiifolia L. (Euphorbiaceae), popularly known as cotton-leaf physicnut, is a milky shrub notable for its medicinal properties. The present study aimed to evaluate the toxic, cytotoxic and genotoxic effects of the latex of J. gossypiifolia, using Allium cepa L. as test system. Seeds of A. cepa were exposed to five concentrations of the latex (1.25; 2.5; 5; 10 and 20 mL/L) in order to evaluate parameters of toxicity (evaluation of root growth), cytotoxicity (mitotic index frequency) and genotoxicity (frequency of chromosome alterations). The latex showed a significant decrease in root mean growth value as well as mitotic index for the tested concentrations, except for 1.25 mL/L, when compared to results from the negative control. The 1.25, 2.5 and 5 mL/L concentrations induced significant chromosome adherences, C-metaphases and/or chromosome bridges, as genotoxic effects. The significant frequency of chromosome bridges also indicated mutagenic potential for chromosomes of J. gossypiifolia as discussed in the paper. Considering that the latex is used in popular therapies, and that the test system A. cepa presents good correlation with tests carried out in mammals, it can be pointed out that its use for medicinal purposes may be harmful to human health especially if ingested.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq