Nuclear DNA content of thirty species of neotropical fishes

The present paper reports nuclear DNA content in 30 Neotropical freshwater fish species and summarizes the data on other Neotropical species presented in the literature. Among Neotropical fishes, the nuclear DNA content ranges from 1.04 ± 0.09 pg/nucleus in Corydoras cf. simulatus (2n = 62) to 248.0 pg/nucleus in Lepidosiren paradoxa (2n = 38). A general analysis of the data obtained in the present study for each species showed that DNA measurements were practically constant at the individual level, while significant differences were observed among individuals of the same population. This observation was valid for all species analyzed and was more evident in those species that presented other karyotypic particularities such as sex chromosomes or supernumerary chromosomes. The importance of changes in nuclear DNA content in the evolutionary process of Neotropical fishes is discussed.

Methodology for kariological study of Brazilian Palms

In the laboratory of cytogenetics of the DBAA-UNESP we are studying the karyotipe of some Brazilian Palms. To determine the best protocol, methods of seed germination, inhibition of mitosis, time to pick up the roots and staining were analyzed. The results shown that the seed germination in sphagnum is effective to achieve good roots. The best time to collect the root tips is between 11 to 12 AM., when there are more cell metaphases. The inhibition of mitosis cycle at metaphases may be effective both with 8-hydroxiquinoleine (0,03% -5 hours) or with cold water (0°C - 18-20 hours). The staining with Giemsa 2% showed the best chromosome figures in the metaphases. Now, to get good metaphases slides the method in use in the lab is: 1) seed germination in sphagnum at room temperature and high humidity; 2) The major roots are cut maintaining at least 5 cm, because this technic allows new emergence of roots, increasing the number of roots collected per germinated seed, that is very important in some species with poor germination rates; 3) To get the mitosis inhibition we are using cold water (0°C) treatment for 18-20 hours, following the standard protocols for conservation and hydrolysis; or enzyme digestion with pectinasecellulase 4) the staining procedures are made using Giemsa 2%. The Brazilian palms species studied and their respective chromosome number were: Aiphanes acanthophylla (2n=30), A. caryotaefolia (2n=30), Syagrus quinquifaria (2n=32), S. coronata (2n=32), S. romanzoffiana (2n=32), Euterpe edulis (2n=36), E. oleracea (2n=36), Copernicia prunifera (2n=36), Scheelea lauromuelleriana (2n=32) and Bactris gasipaes (2n=30).

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 seven species of Eucalyptus L'Hér. (Myrtaceae)

Seven species of the genus Eucalyptus were studied cytogenetically (E. deanei, E. dunni, E. grandis, E. maculata E. propinqua, E. saligna and E. tereticornis). The species showed a symmetrical karyotype with 2n=22 chromosomes, with chromosome length ranging from 0.58 μm to 1.39 μm. Karyotypic analysis indicated homogeneity of morphology and of chromosome number for most of the species of this genus studied here, although casual disploid species with 2n=24 have been found in previous studies. According to these data, a basic number of x=11 was established for this genus. The evolutionary tendency probably occurred by structural alterations (deletions, duplications, additions and translocations) and in some cases by aneuploid chromosome alterations.

Intrachromosomal distribution of telomeric repeats in Eumops glaucinus and eumops perotis (Molossidae, Chiroptera)

The location of chromosomal telomeric repeats (TTAGGG)(n) was investigated in two species of the Molossidae family, Eumops glaucinus and Eumops perotis. The diploid chromosome number (2n) is 40 in E. glaucinus and 48 in E. perotis and the fundamental numbers (FN) are 64 and 58, respectively. It has been suggested that the E. glaucinus karyotype has evolved from the E. perotis karyotype through Robertsonian fusion events. In the present study, the telomeric sequences were detected at the termini of chromosomes in both species. In addition, E. glaucinus also displayed telomeric repeats in centromeric and pericentromeric regions in almost all biarmed chromosomes. Conversely, in E. perotis pericentromeric signals were only observed in two biarmed chromosomes. In both E. glaucinus and E. perotis, such telomeric sequences were observed as part of the heterochromatin. The interstitial sites of telomeric sequences suggest that they are remnants of telomeres of ancestral chromosomes that participated in the fusion event.

Morphologically differentiated sex chromosomes in neotropical freshwater fish

A general survey of the occurrence of morphologically differentiated sex chromosomes in the neotropical freshwater fishes is presented. The total number of 32 occurrences involving simple XX-XY and ZZ-ZW, and multiple X1X2Y, XY1Y2 and ZW1W2 sex chromosome systems is described, with comments on the aspects of sex chromosome evolution in this fish fauna. The occurrence of different sex chromosome systems in related species of the same genus, or in different populations of the same nominal species, involving male and sometimes female heterogamety, and differences in the molecular composition of sex-linked heterochromatin, are considered as indicative of the early stage of sex chromosomes evolution in fish.

Paternity study of an interspecific natural hybrid of the genus Eucalyptus L'Hér (Myrtaceae) based on cytogenetic data

Three species of the genus Eucalyptus (E. dunni, E. grandis, E. saligna) and interspecific hybrid were studied cytogenetically. The Eucalyptus species and the hybrid showed a symmetrical karyotype with 2n=22 chromosomes, with chromosome length ranging from 0.67 to 1.39 μm. Karyotypic analysis indicated a homogenous morphology and chromosome number for the species and the hybrid studied here. Based on the karyotype asymmetry data, together with the chromosome morphology results, the hybrid presented close similarity to E. saligna, suggesting that the latter is one of the parental species involved in the production of the hybrid.

Molecular and Cytogenetic Analyses on Brazilian Youths with Pervasive Developmental Disorders

The Pervasive Developmental Disorders (PDDs) constitute a group of behavioral and neurobiological impairment conditions whose main features are delayed communicative and cognitive development. Genetic factors are reportedly associated with PDDs and particular genetic abnormalities are frequently found in specific diagnostic subgroups such as the autism spectrum disorders. This study evaluated cytogenetic and molecular parameters in 30 youths with autism or other PDDs. The fragile X syndrome was the most common genetic abnormality detected, presented by 1 patient with autism and 1 patient with PPD not-otherwise specified (PPD-NOS). One girl with PDD-NOS was found to have tetrasomy for the 15q11-q13 region, and one patient with autism exhibited in 2/100 metaphases an inv(7)(p15q36), thus suggesting a mosaicism 46,XX/46,XX,inv(7)(p15q36) or representing a coincidental finding. The high frequency of chromosomopathies support the hypothesis that PDDs may develop as a consequence to chromosomal abnormalities and justify the cytogenetic and molecular assessment in all patients with PDDs for establishment of diagnosis.

A radiation hybrid map of bovine X Chromosome (BTAX)

We present a comprehensive radiation hybrid map of the bovine X chromosome (Chr) containing 20 new markers, including both microsatellites and expressed genes. This study was conducted with a 5000-rad whole genome RH cell panel consisting of 90 hybrid cell lines. Retention frequencies of individual markers range from 7.8% for XIST to 31.1% for TGLA325. Statistical analysis with RHMAPPER placed all the loci into five linkage groups under a LOD score criterion of 6.0. These groups could be oriented relative to each other because they included multiple microsatellite loci from the consensus linkage map of the X Chr. Markers included in both this RH map and the bovine cytogenetic map were in a consistent order. The comparative bovine-human map thus generated consists of five blocks of genes, the order of which is conserved, although in the opposite direction when presented as ideograms with p and q arms. Inversions of three blocks account for the difference in gene order across the entirety of the two X Chrs.

Cytogenetic studies in Diplopoda

The Diplopoda have received little attention from cytogeneticists owing mostly to technical difficulties in obtaining mitotic chromosomes, restricting the studies to meiosis and eventual spermatogonial metaphases, which limits the use of modern cytogenetical techniques. A literature search shows that only about 0.1% of all known species have been cytogenetically studied. There are 80,000 species estimated for this group, making it the 3rd. larger class in Arthropoda, after Insecta and Arachnida. The diploid chromosomal number in diplopods varies from 2n=8 to 2n=30 and the sex determination mechanism commonly found is XY/XX. In meiotic prophase, the bouquet formation and the diffuse state in pachytene are typical events. The few works performed on Brazilian fauna add up to 16 species, out of an estimated number of 2000 to 3000 species. The present review reports all the species of diplopods that have been cytogenetically studied so far, each with its chromosome number and sex determination system.

Chromosome analysis of two species of sugar cane pests of the genus Mahanarva (Homoptera, Cercopidae)

Studies on the cytogenetics of Homoptera are scarce. Some references in the literature have reported a chromosome number for the genus ranging from n=5 to 19 and 2n=10 to 2n=39 chromosomes. The genus Mahanarva includes two species of agricultural importance as pests of sugar cane culture in Brazil. We report here the first data concerning the chromosome number and morphology of the species Mahanarva fimbriolata and M. posticata. The chromosome number observed for the two species was 2n=19 for males and 2n=20 for females. The sex determining mechanism of these species was of the XX/X0 type (for males and females, respectively), with chromosome X being the smallest in the complement.

First cytogenetic description of the species Rhamdella microcephala (Pisces, Hepapteridae)

Cytogenetic studies were carried out on Rhamdella microcephala collected from the headwaters of the Machado river, a tributary of the Sapucaí river, Minas Gerais, Brazil. Sixteen specimens exhibited 2n=56 chromosomes, with 18 metacentrics, 30 submetacentrics and 8 subtelocentric-acrocentrics (FN=84). The nucleolar organizing regions (NORs) were identified in the interstitial position on the long arm of a submetacentric pair (pair 12). Chromomycin A3 staining evidenced only the NOR-bearing segments. Positive C-band segments were identified in a pericentromeric position in most of the chromosomes and the NOR-bearing segments were also C-band positive. Some aspects related to the chromosomal characteristics of Rhamdella microcephala are discussed.

Morphologic and Cytogenetic Description of the Small Red Brocket (Mazama bororo Duarte, 1996) in Brazil

We evaluated morphological and cytogenetic data for three animals similar to the species of deer previously described as the Small Red Brocket (Mazama bororo Duarte, 1996). We compared these animals with five M. americana, five M. nana and three hybrids between M. americana and M. nana. The M. bororo chromosomes can be standardized as follows : 8 group A chromosomes (large bi-armed) ; 2 group C chromosomes (small bi-armed) ; 4 group D chromosomes (large acrocentric) ; and 18 group E chromosomes (small acrocentric). There were great differences between this karyotype and those of M. nana and M. americana. With respect to external morphology, the animals in the present study had some similarities to M. americana and M. nana and great similarities with their hybrids. Most of the body measurements of M. bororo were significantly different from those of M. americana and M. nana, but similar to those of the hybrids. Mazama bororo is distributed in the last remnants of the Atlantic forest, extending from the southeastern part of the State of São Paulo to the northeastern part of the State of Paraná, Brazil. The rapid destruction of the Atlantic Forest requires urgent conservation measures for the species.

Chromosome localization of the ribosomal genes 18S and 5S in four stocks of rainbow trout (Oncorhynchus mykiss) from cultivated and naturalized stocks in Brazil

In the present study, fluorescence in situ hybridization (FISH) was employed to determine the chromosomal location of genes 18S rDNA and 5S rDNA in four rainbow trout stocks. In specimens from the stocks of Núcleo Experimental de Salmonicultura de Campos do Jordão and Gavião river, 18S genes were located at a subterminal position in the long arms of two submetacentric chromosomes, whereas in specimens from stocks of Mount Shasta and Teresópolis they were found in the short arms. In all analyzed stocks, 5S genes were located in two chromosome pairs. In a subtelocentric pair, 5S genes were present in the short arms and, in the other submetacentric pair, 5S genes were at an interstitial position. In the latter, 18S and 5S genes were contiguous. Taking into account that both 18S and 5S rDNA genes have been localized in the short arm of a submetacentric chromosome in almost all rainbow trout samples so far studied, the presence of such genes in the long arm, as seen in the samples from Núcleo Experimental de Salmonicultura de Campos do Jordão and Gavião river, supports the hypothesis of a pericentric inversion involving this chromosome segment in the ancestor line of these stocks. The observed polymorphism allowed the identification of a very useful genomic marker, and may therefore constitute an important tool in the genetic management of rainbow trout stocks.

Cytogenetic analysis of A-, B-chromosomes and ZZ/ZW sex chromosomes of Characidium gomesi (Teleostei, Characiformes, Crenuchidae)

Different cytogenetic techniques were used to analyze the chromosomes of Characidium gomesi with the main objective of comparing the base composition of ZZ/ZW sex-chromosomes, B-chromosomes and the heterochromatin of A-chromosomes. The results of digestion of chromosomes with AluI restriction endonuclease (RE), silver and CMA3 staining, C-banding and fluorescence in situ hybridization (FISH) with the 18S rDNA probe suggested the existence of compositional differences between the heterochromatin of ZZ/ZW sex-chromosomes, A- and B-chromosomes, and indicated the presence of different numbers and morphology of B-chromosomes in the samples of this population.