Roberto Laridondo Lui

First description of B chromosomes in the family Auchenipteridae, Parauchenipterus galeatus (Siluriformes) of the São Francisco River basin (MG, Brazil)

B chromosomes are considered additional and non-essential; they likely originate from A chromosomes and follow a distinct evolution. In fish, approximately half of the Neotropical species with B chromosomes are Characiformes and 35% are Siluriformes. There has been no report of B chromosomes in Auchenipteridae until this moment. B chromosomes found in a population of Parauchenipterus galeatus from the São Francisco River basin in the state of Minas Gerais (Brazil) were small, metacentric, totally heterochromatic and exhibited intra-individual and inter-individual variation. The diploid number was 58 chromosomes (22 metacentric, 16 submetacentric, 12 subtelocentric and 8 acrocentric). The nucleolar organizing regions were simple and the heterochromatin intercalated in the ribosomal sites, characterized by CMA(3) and DAPI fluorochromes, was of a GC-rich constitution. The 5S rDNA genes were located in an intercalary position in only one chromosome pair. An hypothesis about the origin of the B chromosomes in P. galeatus and a review on B chromosomes in catfish are also presented in this study.

Characterization of invasive fish species in a river transposition region: evolutionary chromosome studies in the genus Hoplias (Characiformes, Erythrinidae)

Karyotypic analyses were performed in fishes from the genus Hoplias (H.malabaricus and H. lacerdae groups) from the São Francisco River basin (Brazil), in an impacted region by a river transposition which altered the local ecology and fish fauna. The karyotypes were investigated using chromosomal markers obtained from classic and molecular cytogenetics (Giemsa, CMA3 and DAPI staining, C-banding, Ag-NORs, and FISH with 18S rDNA, 5S rDNA and 5SHindIII satellite DNA probes). Two karyotypic forms were found for the H. malabaricus group—karyomorph F, corresponding to the native form from the São Francisco River basin, and karyomorph A, corresponding to the invading form from the Upper Paraná River basin. Specimens from the H. lacerdae group exhibited striking chromosome differences in relation to the H. malabaricus group, thereby enabling good cytotaxonomic characterization and inferences regarding the karyotype evolution of these groups.

Propidium iodide for making heterochromatin more evident in the C-banding technique

Abstract The detection of regions of heterochromatin has been the subject of intense investigation. We investigated an adaptation of the commonly used technique by replacing the nonfluorescent dye, Giemsa, by a fluorescent one, propidium iodide. This adaptation produces greater contrast of the heterochromatic bands in metaphase chromosomes and can be especially valuable when the organisms studied possess heterochromatin that is pale and difficult to visualize. We discuss the interactions of these two dyes with DNA and the excitation of the fluorescent dye when irradiated with ultraviolet light.

Karyotypic diversity between allopatric populations of the group Hoplias malabaricus (Characiformes: Erythrinidae): evolutionary and biogeographic considerations

Three populations of the group Hoplias malabaricus from the hydrographic basins of the Sao Francisco, Araguaia/Tocantins and Xingu Rivers in Brazil were analyzed using classic cytogenetic methods (Giemsa staining, C-banding and Ag-NORs) and molecular methods (fluorescent in situ hybridization with 18S rDNA, 5S rDNA and 5SHindIII satellite DNA probes). The chromosome markers allowed the characterization of these populations as belonging to karyomorph A and the detection of inter-population divergences. These differences likely stem from different evolutionary histories resulting from geographic isolation between populations associated to the dispersive mode of these organisms, reinforcing genetic diversity in the group Hoplias malabaricus.

Comparative Cytogenetics of Giant Trahiras Hoplias aimara and H. intermedius (Characiformes, Erythrinidae): Chromosomal Characteristics of Minor and Major Ribosomal DNA and Cross-Species Repetitive Centromeric Sequences Mapping Differ among Morphologically Identical Karyotypes

Karyotype and cytogenetic characteristics of 2 species of giant trahiras, Hopliasintermedius, São Francisco river basin, and Hopliasaimara, Arinos river (Amazon basin), were examined by conventional (C-banding, Ag-NOR, DAPI/CMA3 double-staining) and fluorescent in situ hybridization (FISH) with 5S, 18S rDNA probes and cross-species Cot-1 DNA probing. Both species invariably had diploid chromosome number 2n = 50 and identical karyotypes composed of 10 pairs of metacentric and 15 pairs of submetacentric chromosomes. On the other hand, staining with base-specific fluorochromes (CMA3, DAPI) and FISH mapping of repetitive DNA sequences showed extensive interspecific differences: while the genome of H. aimara had one submetacentric pair bearing CMA3-positive (DAPI-negative) sites, that of H. intermedius had 4 such pairs; while FISH with a 5S rDNA probe showed one (likely homologous) signal-bearing pair, that with 18S rDNA displayed one signal-bearing pair in H. intermedius and 2 such pairs in H. aimara. Cross-species FISH probing with Cot-1 DNA prepared from total DNA of both species showed no signals of Cot-1 DNA from H. aimara on chromosomes of H. intermedius but reciprocally (Cot-1 DNA from H. intermedius on chromosomes of H. aimara) displayed signals on at least 4 chromosome pairs. Present findings indicate (i) different composition of repetitive sequences around centromeres, (ii) different NOR phenotypes and (iii) distinct taxonomic status of both giant trahira species.

A new technique for obtaining mitotic chromosome spreads from fishes in the field

This study presents an adaptation of current methodologies for preparing mitotic chromosomes from fishes, optimized for use in the field. The high-quality preparations obtained using this modified methodology is suitable for subsequent chromosomal analysis. Importantly, this method is particularly useful when specimen collection sites are far from research laboratories or when researchers are working with highly sensitive species that do not survive long outside of their natural habitats.

Comparative cytogenetics of three populations from the Rhamdia quelen species complex (Siluriformes, Heptapteridae) in two Brazilian hydrographic basins

Abstract Among the more than 30 families of the order Siluriformes, Heptapteridae is composed of 189 species distributed into 24 genera. Rhamdia, which has wide distribution throughout the Neotropical region, presents only 11 valid species, with 8 being in the Brazilian territory. Rhamdia quelen is the only species considered as widely distributed in almost all Brazilian hydrographic basins. It is a Neotropical fish species with a confusing taxonomical history. Classic and molecular cytogenetics data for three populations of this species from two large Brazilian hydrographic basins (Paraná and Araguaia) are presented here. The diploid number found for the three analyzed populations was 58 chromosomes, but with distinct karyotypic formulae. The presence of B chromosomes was detected in the two Araguaia River populations with intra- and interindividual variation. C-banding evidenced little heterochromatin in the three analyzed populations. FISH with 18S rDNA probes evidenced a single chromosome pair bearing this site, confirming the presence of simple NORs, as visualized through silver nitrate staining. The site of 5S rDNA was observed in only one pair of chromosomes, but differing in the marked pair and their location. Based in the differences of the karyotypic formulae and rDNA 5S found between populations on this study and many others available in the literature, it is suggested that this group represents a species complex, and that a new and detailed taxonomical review is necessary.

The role of the Robertsonian rearrangements in the origin of the XX/XY1Y2 sex chromosome system and in the chromosomal differentiation in Harttia species (Siluriformes, Loricariidae)

Harttia is a genus of the subfamily Loricariinae that posses a broad chromosomal variation. In addition to interspecific karyotype diversity within this group, a multiple sex chromosome system, XX/XY1Y2, has been described for Harttia carvalhoi. Thus, this study aimed to determine the role of chromosomal rearrangements in karyotype differentiation in Harttia by classical and molecular cytogenetic procedures. The results show that Robertsonian rearrangements have a prominent role in the chromosomal diversification of the species analysed, which initially leads to hypothesize a diploid number reduction in Harttia torrenticola and H. carvalhoi. The metacentric chromosome 1, shared between H. torrenticola and H. carvalhoi, could have originated from centric fusions from the ancestral karyotype. A centric fission event associated with the first metacentric pair allowed for the origination of a multiple sex chromosome system XX/XY1Y2, specific to H. carvalhoi. This study highlights the relevance of Robertsonian rearrangements in karyotypic differentiation of the species studied and demonstrates that the occurrence of a centric fission, as opposed to a previously hypothesised chromosome fusion, is directly implicated in the origin of the sex chromosome system of H. carvalhoi.

Physical mapping of (GATA)n and (TTAGGG)n sequences in species of Hypostomus (Siluriformes, Loricariidae).

Hypostomus, a genus of Loricariidae, is one of the most diversified and widely distributed genera among fishes of South America (Weber 2003). It is the dominant genus of armoured catfish in Brazilian rivers (Weber 2003). Chromosomally, this group is highly diversified and studies of repetitive DNAs in these species are still in preliminary stages. Hypostomus karyotypes range from 54 chromosomes in H. plecostomus (Muramoto et al. 1968) to 84 chromosomes in Hypostomus sp. 2 (Cereali et al. 2008). Artoni and Bertollo (2001) consider the diploid number of 54 chromosomes as basal for Hypostominae, using Trichomycteridae as outgroup, suggesting that the chromosome evolution of Hypostomus occurred through centric fissions. Recently, Bueno et al. (2012) showed that other chromosomal rearrangements, such as inversions, deletions, duplications and heterochromatinization, could contribute to the chromosomal differentiation within the genus. Repetitive DNAs arranged in tandem are important tools in studies of taxonomic and evolutionary problems. These sequences could be associated with chromosomal rearrangement events (Wichman et al. 1991; Rosa et al. 2012; and others). According to Wichman et al. (1991), satellite DNAs rapidly diverge during evolution. Thereby, they are important for solving the taxonomic and evolutionary problems among related species.

The role of chromosomal fusion in the karyotypic evolution of the genus Ageneiosus (Siluriformes: Auchenipteridae)

Ageneiosus is the most widely distributed genus of the family Auchenipteridae among South American river basins. Although chromosome studies in the family are scarce, this genus has the largest number of analyzed species, with 2n = 54 to 56 chromosomes, differing from the rest of the family (2n = 58). This study aimed to analyze Ageneiosus inermis from the Araguaia River basin. The diploid number found was of 56 chromosomes. Heterochromatin was allocated in terminal region of most chromosomes, plus a pericentromeric heterochromatic block in pair 1, a pair distinguished by size in relation to other chromosomes pairs. AgNORs were detected in only one submetacentric chromosome pair, which was confirmed by FISH. 5S rDNA was present in only one metacentric chromosome pair. Hybridization with [TTAGGG]n sequence marked the telomeres of all chromosomes, in addition to an ITS in the proximal region of the short arm of pair 1. The repetitive [GATA]n sequence was dispersed, with preferential location in terminal region of the chromosomes. Ageneiosus has a genomic organization somewhat different when compared to other Auchenipteridae species. Evidences indicate that a chromosomal fusion originated the first metacentric chromosome pair in A. inermis, rearrangement which may be a basal event for the genus