Estudo citogenético em equinos para detecção de portadores de monossomia do cromossomo X

A cytogenetic study of 22 mares with fertility has showed that four of them had 63,X/64,XX mosaicism. The X-chromosome has presented the expected interstitial heterochromatic C - banding located in the long arm, besides of the usual centromeric band. The great variation of clinic signs observed in mares with mosaicism, could be due to the type of zygote or the time the mosaicism occured.

A LINE2 repetitive DNA sequence from the cichlid fish, Oreochromis niloticus: sequence analysis and chromosomal distribution

We report the cloning and characterization of a long interspersed nucleotide element (LINE) fi-om a cichlid fish, Oreochromis niloticus, and show the distribution of this element, called CiLINE2 for cichlid LINE2, in the chromosomes of this species. The identification of an open reading frame in CiLINE2 with amino acid sequence similarity to reverse transcriptases encoded by LINE-like elements in Caenorhabditis elegans, Platemys spixii, Schistosoma mansoni, Gallus gallus (CRI), Drosophila melanogaster (I factor), and Homo sapiens (LINE2), as well as the structure of the element, suggest it is a member of this family of non-long terminal repeat-containing retrotransposons. Search of a DNA sequence database identified sequences similar to CiLINE2 in four other fish species (Haplotaxodon microlepis, Oreochromis mossambicus, Pseudotropheus zebra, and Fugu rubripes). Southern blot hybridization experiments revealed the presence of sequences similar to CiLINE2 in all Tilapiini species analyzed from the genera Oreochromis, Tilapia, and Sarotherodon, and gave an estimated copy number of about 5500 for the haploid genome of O. niloticus. Fluorescent in situ hybridization showed that CiLINE2 sequences were organized in small clusters dispersed over all chromosomes of O. niloticus, with a higher concentration near chromosome ends. Furthermore the long arm of chromosome 1 was strikingly enriched with this sequence. The distribution of LINE2-related elements might underlie the difference in chromosome banding patterns observed between cold-blooded vertebrates and mammals.

Intrageneric Karyotypic Variation in Pseudopaludicola (Anura: Leiuperidae) and Its Taxonomic Relatedness

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Chromosome analysis in Pseudopaludicola (Anura, Leiuperidae), with description of sex chromosomes XX/XY in P-saltica

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Constitutive heterochromatin, G-bands and nucleolus-organizer regions in four species of Didelphidae (Marsupialia)

Chromosomes of Didelphis albiventris, D. marsupialis, Philander opossum and Lutreolina crassicaudata, four species of marsupials with very similar karyotypes and 2n=22 were studied. All the chromosomes were acrocentrics except the X in L. crassicaudata, which is a metacentric. The G-band patterns of these species are similar but the distribution of constitutive heterochromatin differs among them as shown by C-banding. The hypothesis that the X in L. crassicaudata might be an isochromosome derived from the acrocentric X in the other species is discarded since G-and C-banding patterns differ in the two arms. In D. marsupialis the Ag-NORs are terminal and located in both arms of one pair and in the long arms of two pairs of medium-sized autosomes. In P. opossum the NOR-bearing chromosomes could be precisely identified through simultaneous silver staining and G-banding. The Ag-NORs are terminal and located at the short arm of pair 5 and the long arm of pair 7. © 1982 Dr W. Junk Publishers.

Characteristics of NOR-banded chromosomes of Mangalarga horses

The nucleolar organizer regions (NORs) of Mangalarga horses were characterized by analysis of NOR-banded metaphase chromosomes according to the technique of Goodpasture and Bloom (Chromosoma 53: 37-50, 1975). NOR banding was detected by silver staining in the telomeric region of the short arm of pair no. 1, in the region adjacent to the centromere of the long arm of pair no. 25 and in the proximal region of the long arm of pair no. 31. Associations of NOR-bearing chromosomal regions occurred in 12% of all metaphases and were frequent between the chromosomes of pair no. 1. Most (52.15%) of the NOR bands appeared on four chromosomes in both males and females. The maximum number of NOR-banded chromosomes was six, though only 11.34% of the cells examined showed this characteristic.

Fish cytogenetic research: Advances, applications and perspectives

Methods developed since 1976 for harvesting, preparing and banding fish chromosomes are now commonly used for taxonomic and phylogenetic studies, genetic control and chromosome manipulations in fish breeding and in monitoring aquatic pollutants by examining chromosomal aberrations. These studies have chiefly concerned common temperate freshwater species; the same procedures, when applied to marine and coldwater fish, often provide unsatisfactory results, especially in cell culture. A concerted effort should be made in marine fish, and to develop molecular cytogenetic methods to provide a more powerful tool to study chromosomal evolution. © 1991 BRILL.

Fluorescence in situ hybridization with rDNA probes on chromosomes of two nucleolus organizer region phenotypes of a species of Eigenmannia (Pisces, Gymnotoidei, Sternopygidae)

Nucleolus organizer regions (NORs) were analysed in two related and geographically close populations of Eigenmannia sp.1 (Pisces, Gymnotoidei, Sternopygidae) using silver staining and fluorescence in situ hybridization (FISH). The two populations differed in their AS-NOR phenotypes, displaying fixed differences in the NOR-bearing chromosome pairs. FISH with rDNA probes showed that these differences were due to the location of rDNA cistrons. This finding, showing fixed NOR differences between two populations belonging to the same species in a connected river system, is highly significant in terms of evolutionary change, possibly indicating an initial step of genetic differentiation. This result also has important implications from the cytosystematic point of view, as NORs usually have a very constant karyotypic location in fish species and have been used as species-specific chromosome markers.

Use of lipopolysaccharides to characterize Bradyrhizobium spp.

Three methods of extraction of lipopolysaccharide (LPS) were compared-the conventional hot phenol-water method with 40% phenol, a modified form of this method using 10% phenol, and the hot saline method. Good recovery of LPS was achieved by each of the three methods, with the LPS found in the aqueous phase with the two phenol-based procedures. The application of SDS-PAGE to the LPS extracts, followed by silver staining, showed similar banding with all three methods of extraction. When the hot saline extraction LPS fraction from eight strains of Bradyrhizobium spp. and eight strains of Bradyrhizobium japonicum was compared with SDS-PAGE, characteristic profiles were achieved. Serological analysis of eight strains of Bradyrhizobium spp., using antisera prepared against whole cells in agglutination reactions, showed extensive sharing of antigens. When antisera was prepared using outer membrane LPS, extracted by the hot saline method, the amount of cross-reaction was reduced greatly. The results indicated that LPS provide an efficient means of obtaining monospecific antisera to be used for serological identification of strains of Bradyrhizobium spp. and that the hot saline extraction method is recommended for a fast, simple and efficient way to obtain LPS and characterize this bacterium.

A cytogenetic study of Diplomystes mesembrinus (Teleostei, Siluriformes, Diplomystidae) with a discussion of chromosome evolution in siluriforms

The mitotic chromosomes, nucleolus organizer regions (NORs), C-banding pattern and nuclear DNA content of Diplomystes mesembrinus were studied. The karyotype, with 2n=56 chromosomes (22m+24sm+6st+4a), has a high chromosome arm number (NF = 102), one chromosome pair with NORs, and a very small amount of heterochromatin. The NOR-bearing arm is entirely heterochromatic and exhibits a marked size polymorphism. The diploid DNA content detected in erythrocyte nuclei of D. mesembrinus was 2.57 ± 0.15 pg/nucleus. The chromosome evolution in Siluriformes is discussed on the basis of available cytogenetic data and it is proposed that 2n=56 is synapomorphic for the order.

Cytogenetic analysis of Epicauta atomaria (Meloidae) and Palembus dermestoides (Tenebrionidae) with Xyp sex determination system using standard staining, C-bands, NOR and synaptonemal complex microspreading techniques

The mitotic and meiotic chromosomes of the beetles Epicauta atomaria (Meloidae) and Palembus dermestoides (Tenebrionidae) were analysed using standard staining, C-banding and silver impregnation techniques. We determine the diploid and haploid chromosome numbers, the sex determination system and describe the chromosomal morphology, the C-banding pattern and the chromosome(s) bearing NORs (nucleolar organizer regions). Both species shown 2n = 20 chromosomes, the chromosomal meioformula 9 + Xyp, and regular chromosome segregation during anaphases I and II. The chromosomes of E. atomaria are basically metacentric or submetacentric and P. dermestoides chromosomes are submetacentric or subtelocentric. In both beetles the constitutive heterochromatin is located in the pericentromeric region in all autosomes and in the Xp chromosome; additional C-bands were observed in telomeric region of the short arm in some autosomes in P. dermestoides. The yp chromosome did not show typical C-bands in these species. As for the synaptonemal complex, the nucleolar material is associated to the 7th bivalent in E. atomaria and 3rd and 7th bivalents in P. dermestoides. Strong silver impregnated material was observed in association with Xyp in light and electron microscopy preparations in these species and this material was interpreted to be related to nucleolar material.

Cytogenetics of two species of Paratelmatobius (Anura: Leptodactylidae), with phylogenetic comments

In this paper we provide a cytogenetic analysis of Paratelmatobius cardosoi and Paratelmatobius poecilogaster. The karyotypes of both species showed a diploid number of 24 chromosomes and shared some similarity in the morphology of some pairs. On the other hand, pairs 4 and 6 widely differed between these complements. These karyotypes also differed in their NOR number and location. Size heteromorphism was seen in all NOR-bearing chromosomes of the two karyotypes. In addition, both karyotypes showed small centromeric C-bands and a conspicuous heterochromatic band in the short arm of chromosome 1, although with a different size in each species. The P. cardosoi complement also showed other strongly stained non-centromeric C-bands, with no counterparts in the P. cardosoi karyotype. Chromosome staining with fluorochromes revealed heterogeneity in the base composition of two of the non-centromeric C-bands of P. cardosoi. Comparison of the chromosomal morphology of these Paratelmatobius karyotypes with that of P. lutzii showed that the P. poecilogaster karyotype is more similar to that of P. lutzii than P. cardosoi. These cytogenetic results agree with the proposed species arrangements in the P. cardosoi and P. lutzii groups based on morphological and ecological data.

Evaluation of naturally and artificially aged seeds of Phaseolus vulgaris L.

Seed ageing is natural phenomenon that occurs in all seeds, including those stored in dry and low temperature rooms. Phaseolus vulgaris L. cv. 'Carioca' seeds harvested in five different years were stored in at 15°C. Seed samples were germinated and evaluated according to The Brazilian Rules for Testing Seeds. The newest seed sample was submitted to artificial ageing. Small (∼1g) samples of all materials (naturally and artificially aged) were ground and proteins extracted. Equal quantities of protein were loaded onto the gels and electrophoresis carried out. Although seeds submitted to 24, 48, 72 and 96 hours of artificial ageing did not show any statistical difference (5%) in relation to unaged seeds in physiological parameters related to emergence, some statistically different parameters related to dry matter and length of shoots and roots occurred. A 24 h of artificial ageing at 41°C improved these factors. Protein patterns were changed after 72 h of artificial ageing and naturally aged seeds showed alterations after two years storage at 15°C. Aged seeds, naturally and artificially, had decreasing germination, vigour, and changes in the characteristic banding pattern of proteins. Physiological parameters and electrophoresis examination showed that for naturally aged seeds physiological parameters were more sensitive while artificially aged seeds, (41°C/100% RH), had electrophoretic profiles that were more efficient for seed lot discrimination.