Application of C-banding in two Arachis wild species, Arachis pintoi Krapov. and W.C. Gregory and A-villosulicarpa Hoehne to mitotic chromosome analyses

Two wild diploid (2n = 20 chromosomes) and self-pollinating Arachis species, Arachis Pintoi Krapov and W.C.Gregory and A. villosulicarpa Hoehne were submmited to C-band technique to karyotype analyses. Root tips were employed in the analyses. Morphometric data chose that chromosome lengths varied from 3.12 in A. villosulicarpa to 1.45 in A. Pintoi. Karyotype formula obtained was 10sm to A. Pintoi and 9sm + 1m to A. villosulicarpa. There was a predominance of pericentromeric C-band in all mitotic metaphasic chromosomes in both species. Besides C-band values, both species still did not differ in respect to chromosome absolute and relative lengths, centromeric index, symmetry index and total karyotype haploid length. C-band and morphometric data did not show strong or significant differences which could separate these two species of peanut which belong to evolutive different sections.

Molecular cytogenetic analysis of heterochromatin in the chromosomes of tilapia, Oreochromis niloticus (Teleostei : Cichlidae)

The structure of the heterochromatic bands in mitotic chromosomes of the important tropical aquaculture species of tilapia, Oreochromis niloticus, was investigated by the combination of the C-banding technique, chromosomal digestion with two restriction endonucleases and fluorescence in situ hybridization (FISH) of two satellite DNAs (SATA and SATB). The tilapia chromosomes presented heterochromatic bands in the centromeres and in the short arms of almost all chromosomes that were differentially digested by the restriction endonucleases HaeIII and EcoRI. FISH of SATA showed that the satellite sequence is distributed in the centromeric region of all chromosomes of tilapia. FISH also revealed an intense hybridization signal for SATB in only one chromosome pair, but less intense signals were also present in several other pairs. The digestion of tilapia chromosomes by HaeIII and EcoRI was positively correlated with the position of SATA and SATB in chromosomes as revealed by FISH. The results obtained may be useful in future molecular and genetic studies of tilapias.

Phosphorylation of Histone H3S10 in Animal Chromosomes: Is There a Uniform Pattern?

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Application of C-banding in two Arachis wild species, Arachis Pintoi Krapov. and W.C. Gregory and A. villosulicarpa Hoehne to mitotic chromosome analyses

Two wild diploid (2n = 20 chromosomes) and self-pollinating Arachis species, Arachis Pintoi Krapov and W.C. Gregory and A. villosulicarpa Hoehne were submmited to C-band technique to karyotype analyses. Root tips were employed in the analyses. Morphometric data chose that chromosome lengths varied from 3.12 in A. villosulicarpa to 1.45 in A. Pintoi. Karyotype formula obtained was 10sm to A. Pintoi and 9sm + 1m to A. villosulicarpa. There was a predominance of pericentromeric C-band in all mitotic metaphasic chromosomes in both species. Besides C-band values, both species still did not differ in respect to chromosome absolute and relative lengths, centromeric index, symmetry index and total karyotype haploid length. C-band and morphometric data did not show strong or significant differences which could separate these two species of peanut which belong to evolutive different sections.

Cytogenetic studies of Mastacembelus armatus (Pisces, Mastacembelidae)

The mitotic chromosomes, nucleolus organizer regions (NORs), C-banding pattern and nuclear DNA content of Mastacembelus armatus were studied. The karyotype (2n = 48; 10m + 6sm + 4st + 28a) was characterized by the presence of one chromosome pair with NORs and a small quantity of heterochromatin. The DNA content observed in erythrocyte nuclei of M. armatus was 1.39 +/- 0.08 pg. Comparison of this karyotype with those of other Synbranchiformes revealed a strong similarity, suggesting that small chromosome rearrangements may have been maintained during its evolutionary history.

Variability of 18S rDNA locus among Symphysodon fishes: chromosomal rearrangements

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Chromosomal polymorphism in 12 populations of Mikania micrantha (Compositae)

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

HETEROCHROMATIN AND KARYOTYPE REORGANIZATION IN FISH OF THE FAMILY ANOSTOMIDAE (CHARACIFORMES)

Mitotic chromosomes of four fish species of the family Anostomidae, belonging to the genera Leporinus, Leporellus, and Schizodon, were studied. With 2n = 54 meta- and submetacentric chromosomes, this family appears to be characterized by marked karyotypic stability. Although perceptible differences exist, mainly in the amount of constitutive heterochromatin present in the chromosomes of these species, these differences do not affect the structure and/or size of these chromosomes. Chromatin substitutions and/or modifications may have led, in one direction, to an increase in heterochromatin in some species and, in the opposite direction, to heterochromatin reduction in others. Whether these changes are accompanied by changes in the amount of euchromatin in the chromosomes is an open question. The nucleolar organizer regions, which may be located on different chromosomes in the various species, may also be indicators of reorganization of these karyotypes.

Cytogenetics of three Brazilian Goniosoma species: A new record for diploid number in Laniatores (Opiliones, Gonyleptidae, Goniosomatinae)

Currently, 60 species of harvestmen have been karyotyped and all of these are from the Nearctic and Palearctic regions. This work is the first cytogenetic report of three gonyleptid species of the suborder Laniatores: Goniosoina aff. badiuln, G. proxiinuni and G. spelaeum of the Neotropical region, from the southeastern region of Brazil. Conventional Giemsa stain chromosome preparations were obtained from embryonic cells and adult male testes. Embryo mitotic plates of G. aff. badium and G. proximum indicated 88 chromosomes, and mitotic spermatogonial plates of G. spelaeum males revealed intra- and interindividual variation of chromosome number, ranging from 92-109 chromosomes. In the three analyzed species, the mitotic chromosomes were meta- or subinetacentric with no obvious sex chromosomes being identified during mitosis. Prophase I spermatocytes of G. spelaeum also revealed intra- and interindividual bivalent number variation and furthermore indicated the presence of multivalence. The karyotypes of these three Goniosoina species exhibited the largest chromosome pair with a negative heteropycnosis in the distal region of the shortest arrn chromosomes of G. spelaeum submitted to silver impregnation evidenced this negative heteropycnotic region as nucleolus organizer region (NOR). These results, when compared with cytogenetic data of other Laniatores species from the Palearctic region, indicated that a new record for diploid chromosome number probably characterize the genus Goniosoma in the Neotropical region.

KARYOTYPE AND NUCLEOLUS ORGANIZER REGIONS OF PYRRHULINA CF AUSTRALIS (PISCES, CHARACIFORMES, LEBIASINIDAE)

The mitotic chromosomes and nucleolus organizer regions (NORs) of the Upper Parana Basin fish Pyrrhulina cf. australis were studied. The karyotype (2n = 40; 6st + 34a) is characterized by the presence of four chromosome pairs with NORs, one of them with NORs in both terminal regions. Comparison of this karyotype with those of other Characiformes, revealed a strong similarity to the family Erythrinidae.

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 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.

Simple method for culture of peripheral blood lymphocytes of Testudinidae.

We developed and optimized a simple, efficient and inexpensive method for in vitro culture of peripheral blood lymphocytes from the Brazilian tortoise Chelonoidis carbonaria (Testudinidae), testing various parameters, including culture medium, mitogen concentration, mitotic index, culture volume, incubation time, and mitotic arrest. Peripheral blood samples were obtained from the costal vein of four couples. The conditions that gave a good mitotic index were lymphocytes cultured at 37°C in minimum essential medium (7.5 mL), with phytohemagglutinin as a mitogen (0.375 mL), plus streptomycin/penicillin (0.1 mL), and an incubation period of 72 h. Mitotic arrest was induced by 2-h exposure to colchicine (0.1 mL), 70 h after establishing the culture. After mitotic arrest, the cells were hypotonized with 0.075 M KCl for 2 h and fixed with methanol/acetic acid (3:1). The non-banded mitotic chromosomes were visualized by Giemsa staining. The diploid chromosome number of C. carbonaria was found to be 52 in females and males, and sex chromosomes were not observed. We were able to culture peripheral blood lymphocytes of a Brazilian tortoise in vitro, for the preparation of mitotic chromosomes.

A method for chromosome preparations from large fish specimens using in vitro short-term treatment with colchicine

This paper describes a new technique for preparing mitotic fish chromosomes using short-term in vitro treatment with colchicine. The results show that a large number of good quality metaphases (many suitable for chromosome banding) can be obtained by this technique, which requires an average of 1 h and 30 min for all steps. The procedure considerably reduces the time normally required for chromosome preparations in fish.

Complex sex chromosome system in Eigenmannia sp. (Pisces, Gymnotiformes)

Chromosomes of Eigenmannia sp. (7 males and 15 females) collected from the Tietê River in Botucatu (SP, Brazil) were examined from gill, kidney and testicular cells. The diploid chromosome number in males was 2n=31 and in females, 2n=32. In both sexes the number of chromosomal arms was 40. The difference in diploid number was due to the fusion of two acrocentrics. Mitotic and meiotic studies suggested that one of the fused acrocentrics was the Y chromosome. The sex-determining mechanism in Eigenmannia sp. could therefore be XX, AA in the female and X, \-YA A in the males. One of the males presented 2n=30 chromosomes due to the occurrence of another fusion of acrocentrics. C-banding analysis of the mitotic chromosomes revealed constitutive heterochromatin in the centromeric regions of all acrocentrics. However, small metacentrics were C-band negative. The YA chromosome is C-band negative except for a small amount of heterochromatin in the centromeric region. The nucleolar organizer region as identified by Ag-staining is present in the interstitial region of chromosome pair No. 10. © 1984 Dr W. Junk Publishers.