Chromosomal Study of Colostethus brunneus from the Type Locality and Two Related Species (Anura – Dendrobatidae)

In this paper, we report a chromosomal study of three Brazilian species of Colostethus, C. brunneus from the type locality, Colostethus sp. (aff. trilineatus), and Colostethus sp., which is morphologically similar to C. brunneus. The diploid number for C. brunneus was 2n = 24 chromosomes, in agreement with that previously described for specimens from Peru. Colostethus sp. (aff. trilineatus) and Colostethus sp. showed a very similar karyotype with 22 chromosomes. The NOR was located on pair 3 in C. brunneus, on pair 4 in Colostethus sp. (aff. trilineatus), and on pair 2 in Colostethus sp. In one specimen of Colostethus sp., an additional NOR site was located on pair 7 in only one of the homologs. This extra Ag-NOR site was confirmed by FISH using an rDNA probe. In addition to the NOR location, the C-banding pattern was also species-specific, despite the similar chromosomal morphology of the species. These results indicate that although these species may be closely related, there is a clear dichotomy in their chromosome number.     

Karyotypes of two American field crickets: Gryllus rubens and Gryllus sp. (Orthoptera: Gryllidae)

The karyotypes of Gryllus rubens (Scudder) and Gryllus sp. collected in the US were studied using conventional Giemsa staining and two differential staining methods. Both species had a chromosome complement of 2n = 28 + XX/XO, and the X chromosome was large and metacentric. In addition, nucleolus organizer regions (NOR) were detected in the short arm of one pair in the two species, and the NOR showed variation in size. The two species had species‐specific chromosome configuration and C‐banding patterns. In Gryllus sp., the chromosome configuration showed polymorphism in size and type among individuals, and the distal C‐bands of Gryllus sp. were larger than those of G. rubens.     

Chromosome Differentiated Populations of Cruzii: Evidence for a Third Sibling Species

Anopheles cruziiis the most common species of mosquito in Southeast Brazil and a vector of human and monkey malaria. The banding pattern of the ovarian polytene chromosomes and the frequencies of paracentric inversions of individuals from two populations were studied. A new sequence of bands on the sex chromosome, defined as form C, was disclosed. In both populations where forms A (considered as standard) and C are sympatric no heterozygotes were detected. A sequence of events that could account for the observed changes in the banding sequences of the X chromosome forms was proposed. The frequencies of 22 paracentric inversions were used to assess panmixia and the results indicated the presence of two distinct genetic pools in each population. We consider these results as evidence of another sibling species in the taxon cruzii, characterized by a distinctive form of the X chromosome and provisionally designated Anopheles cruziispecies C.     

Karyotypic study of titi monkeys,Callicebus moloch brunneus

A karyotypic study on a subspecies of the dusky titi,Callicebus moloch brunneus, was carried out and a third karyotype ofC. moloch was discovered. The chromosome number of this subspecies is 48. The autosomes consist of 5 subtelocentric, 5 submeta- or metacentric, and 13 acrocentric chromosome pairs. The X chromosome and the Y chromosome are submetacentric and metacentric, respectively. A comparative study with other subspecies of theC. moloch group (i.e.,C. m. cupreus andC. m. ornatus with 2n=46 andC. m. donacophilus with 2n=50) suggests that the karyotype ofbrunneus occupies a position intermediate between the two other karyotypes ofC. moloch, but nearer to that of 2n=50. The presumed total differences betweenbrunneus andcupreus comprise one Robertsonian rearrangement, one centromeric transposition and four pericentric inversions, and those betweenbrunneus anddonacophilus involve one translocation or breakage (possibly corresponding to two events, that is, one Robertsonian rearrangement and one centromeric transposition).     

Chromosomes of thecercopithecus aethiops species group:C. aethiops (Linnaeus, 1758),C. cynosurus (Scopoli, 1786),C. pygerythrus (Cuvier, 1821), andC. sabaeus (Linnaeus, 1766)

The banded karyotypes of 34 monkeys of known geographic origin and belonging to the Cercopithecus aethiops group of species (C. aethiops, C. pygerythrus, C. cynosurus, C. sabaeus) show that chromosome evolution in this group is highly conservative. All species have 2n =60 chromosomes with very similar chromosome banding. However, differences were found both within and between species. A polymorphism of the NOR area of the “marked” chromosome pairs was found in all taxa (9 of 34 animals). All individuals referred to C. sabaeus,from both West Africa and the Barbados, are characterized by having highly positive G- and C- banded terminal sequences on chromosomes 7,10,12, and 14. Outgroup comparisons with other primates and a parsimony analysis suggest that these terminal bands are derived and are probably good taxonomic and phylogenetic indicators. Moreover, chromosome 18 is variable both between and within species in G banding and in short-arm length. The existence of within-species variation in karyotypes suggests that karyological comparisons must be based on adequate samples that include specimens coming from all the major geographic populations of the species concerned.     

A population genomic scan in Chorthippus grasshoppers unveils previously unknown phenotypic divergence

Understanding the genetics of speciation and the processes that drive it is a central goal of evolutionary biology. Grasshoppers of the Chorthippus species group differ strongly in calling song (and corresponding female preferences) but are exceedingly similar in other characteristics such as morphology. Here, we performed a population genomic scan on three Chorthippus species (Chorthippus biguttulus, C. mollis and C. brunneus) to gain insight into the genes and processes involved in divergence and speciation in this group. Using an RNA‐seq approach, we examined functional variation between the species by calling SNPs for each of the three species pairs and using F_ST‐based approaches to identify outliers. We found approximately 1% of SNPs in each comparison to be outliers. Between 37% and 40% of these outliers were nonsynonymous SNPs (as opposed to a global level of 17%) indicating that we recovered loci under selection. Among the outliers were several genes that may be involved in song production and hearing as well as genes involved in other traits such as food preferences and metabolism. Differences in food preferences between species were confirmed with a behavioural experiment. This indicates that multiple phenotypic differences implicating multiple evolutionary processes (sexual selection and natural selection) are present between the species.     

Population differences in the expression of nucleolus organizer regions in the grasshopperEyprepocnemis plorans

Summary Fluorescence in situ hybridization revealed the presence of ribosomal RNA genes in paracentromeric regions of all A chromosomes and in the distal half of B chromosomes in embryonic cells from Moroccan specimens of the grasshopperEyprepocnemis plorans. The expression of these genes was monitored by the presence of nucleoli attached to each chromosome bivalent in diplotene cells from males collected from two different Moroccan populations and was compared to previous data of Spanish populations. Whereas only the nucleolus organizer regions (NORs) on S₉–S₁₁ and X chromosomes were active in the Spanish specimens. Moroccan individuals showed NOR activity in all chromosomes. The rRNA genes on the B chromosome were inactive in both populations. The S₉ and S₁₀ NORs were less active in Moroccan specimens than in Spanish specimen, which might be partly explained by the negative interdependence for expression of the S₁₀ NOR with respect to those on L₂ and X chromosomes. On the other hand, the X NOR was more active in Moroccan specimens than in Spanish specimens, and this might be partly due to the positive effect that the presence of B chromosomes has on the expression of this NOR. The implications of these observations on current models of NOR activity regulation are discussed.Abbreviation NOR nucleolus organizer region     

NOR Sites Detected by Ag-DAPI Staining of an Unusual Autosome Chromosome of Bradysia Hygida (Diptera:Sciaridae) Colocalize with C-banded Heterochromatic Region

The study of chromosomes in insects is a good tool in mitotic process analysis, zoographic localization and evolution investigation. Among them, the Sciaridae offers a karyotype with a small number of chromosomes, where the heterochromatin and nucleolar organizer region, NOR, are easily analyzed in metaphase chromosomes obtained from cerebral ganglia squashes. In this work, the heterochromatic regions on Bradysia hygida mitotic chromosomes, revealed by C-banding, were identified as centromeric blocks on A and C chromosomes and as dark interstitial region in B and X chromosomes. By Ag-DAPI staining, active nucleolus organizer region, NOR, was revealed associated to the constitutive heterochromatin in the end of the C autosome chromosome. The C-band regions and the unusual ribosomal site localization are discussed.     

X-linked heterochromatin distribution in the holocentric chromosomes of the green apple aphid <Emphasis Type="Italic">Aphis pomi</Emphasis>

Chromatin organization in the holocentric chromosomes of the green apple aphid Aphis pomi has been investigated at a cytological level after C-banding, NOR, Giemsa, fluorochrome staining and fluorescent in situ hybridization (FISH). C-banding technique showed that heterochromatic bands are exclusively located on X chromosomes. This data represents a peculiar feature that clearly contradicts the equilocal distribution of heterochromatin typical of monocentric chromosomes. Moreover, silver staining and FISH carried out with a 28S rDNA probe localized rDNA genes on one telomere of each X chromosome; CMA₃ staining reveals that these silver positive telomeres are the only GC-rich regions among A. pomi heterochromatin, whereas all other C-positive bands are DAPI positive thus containing AT-rich DNA.     

Localization of 5S rRNA Loci in Three Coregonid Species (Salmonidae)

In the present study the chromosome distribution of the 5S rDNA loci and its relation to the major rDNA genes were investigated in three Coregonid species (Salmonidae): Coregonus lavaretus, Coregonus peled and Coregonus albula, a family which has experienced large karyotype rearrangements along its evolution starting from a tetraploid ancestor. 5S PRINS/CMA₃ sequential staining together with previous data enabled us to locate 5S rRNA genes and nucleolar organizer regions (NORs) in the three species analyzed. PRINS revealed the 5S rDNA cluster at the distal part of the long arm of a similar submetacentric chromosome pair in the three species. Our data indicate that 5S rDNA clusters have probably conserved chromosomal location in the genus Coregonus, whereas 45S rDNA (NOR) sites are clearly differentiated, from a single locus in C. peled, to multiple loci in C. lavaretus and highly polymorphic multichromosomal location in C. albula.     

Accumulation of chloroplast DNA sequences on the Y chromosome of Silene latifolia

Silene latifolia is a model dioecious plant with heteromorphic sex chromosomes. The Y chromosome is the largest in this species. Theoretical models propose an accumulation of repetitive DNA sequences in non-recombining parts of the Y chromosome. In this study, we isolated a BAC7H5 clone preferentially hybridizing to the Y chromosome of S. latifolia. Sequence analysis revealed that this BAC7H5 contains part of the chloroplast genome, indicating that these chloroplast sequences have accumulated on the Y chromosome and also may contribute to its large size. We constructed Y chromosome- and X chromosome-specific libraries and screened them to find Y- and/or X-linked copies of chloroplast sequences. Sequence analysis revealed higher divergence of a non-genic region of the chloroplast sequences located on the Y chromosome while genic regions tested showed only very low (max 0.9%) divergence from their chloroplast homologues.     

Comparative analysis of rDNA location in five Neotropical gomphocerine grasshopper species

We report here, for the first time, the chromosome complement, number and location of the nucleolar organizer regions (NORs) revealed by silver staining (AgNO₃) and fluorescent in situ hybridization (FISH) in five Neotropical gomphocerine species: Rhammatocerus brasiliensis, R. brunneri, R. palustris, R. pictus and Amblytropidia sp. The objective of this study was to summarize available data and propose a model of chromosome evolution in Neotropical gomphocerines. All five species studied showed chromosome numbers consisting of 2n = 23,X0 in males and 2n = 24,XX in females. Amblytropidia sp. was the only species showing a bivalent (M₈) with megameric behavior during meiosis. The rDNA sites were restricted to autosomal pairs, i.e. the pericentromeric region of the S₉ chromosome, the consensus NOR location in all five species. R. brasiliensis was the only species showing additional NORs on M₄ and M₆ pairs which, likewise the S₉ NOR, were active in all cells analyzed. Comparison of these results with those reported previously in Palearctic gomphocerine species suggests higher resemblance of Neotropical species with the Old World species also possessing 23/24 chromosomes. Evolutionary mechanisms responsible for the observed interspecific variation in NOR location in this group are discussed.     

Chromosome banding in Amphibia

The mitotic and meiotic chromosomes of the marsupial frog Gastrotheca riobambae were analysed with various banding techniques. The karyotype of this species is distinguished by considerable amounts of constitutive heterochromatin and unusual, heteromorphic XY sex chromosomes. The Y chromosome is considerably larger than the X chromosome and almost completely heterochromatic. The analysis of the banding patterns obtained with GC- and AT-base-pair-specific fluorochromes shows that the constitutive heterochromatin in the Y chromosome consists of at least three different structural categories. The only nucleolus organizer region (NOR) of the karyotype is localized in the short arm of the X chromosome. This causes a sex-specific difference in the number of NOR: female animals have two NORs in diploid cells, male animals one. No cytological indications were found for the inactivation of one of the two X chromosomes in the female cells. In male meiosis, the heteromorphic sex chromosomes form a characteristic sex-bivalent by pairing their telomeres in an end-to-end arrangement. The significance of the XY/XX sex chromosomes of G. riobambae for the study of X-linked genes in Amphibia, the evolution of sex chromosomes and their specific DNA sequences, and the significance of the meiotic process of sex chromosomes are discussed.     

Cytogenetic study of F1 hybrids betweenCrepis dinarica andCrepis froelichiana (Asteraceae)

Crepis dinarica andC. froelichiana are two closely related species of theC. praemorsa complex. Even though they exhibit the same chromosome number (2n = 8) and similar idiogram shape, they differ widely in quantity and distribution of heterochromatin bands. The hybrids between these two species comprise three morphological types. Parental genomes were distinguished in hybrids by Giemsa differential staining (C-banding). Although meiosis presents only a few abnormalities (about 2.4%), the percentage of aborted pollen grains is very high (90%).     

<Emphasis Type="Italic">Drosophila simulans Lethal hybrid rescue</Emphasis> mutation (<Emphasis Type="Italic">Lhr</Emphasis>) rescues inviable hybrids by restoring X chromosomal dosage compensation and causes fluctuating asymmetry of development

The Drosophila simulans Lhr rescues lethal hybrids from the cross of D. melanogaster and D. simulans. We describe here, the phenotypes of Lhr dependent rescue hybrids and demonstrate the effects of Lhr on functional morphology of the salivary chromosomes in the hybrids. Our results reveal that the phenotypes of the ‘Lhr dependent rescued’ hybrids were largely dependent on the genetic background and the dominance in species and hybrids, and not on Lhr. Cytological examination reveal that while the salivary chromosome of ‘larval lethal’ male carrying melanogaster X chromosome was unusually thin and contracted, in ‘rescued’ hybrid males (C _mel X _mel Y _sim ; A _mel A _sim ) the X chromosome showed typical pale staining, enlarged diameter and incorporated higher rate of ³H-uridine in presence of one dose Lhr in the genome. In hybrid males carrying simulans X chromosome (C _mel X _sim Y _mel ; A _mel A _sim ), enlarged width of the polytene X chromosome was noted in most of the nuclei, in Lhr background, and transcribed at higher rate than that of the single X chromosome of male. In hybrid females (both viable, e.g., C _mel X _mel X _sim ; A _mel A _sim and rescued, e.g., C _mel X _mel X _mel ; A _mel A _sim ), the functional morphology of the X chromosomes were comparable to that of diploid autosomes in presence of one dose of Lhr. In hybrid metafemales, (C _mel X _mel X _mel X _sim ; A _mel A _sim ), two dose of melanogaster X chromosomes and one dose of simulans X chromosome were transcribed almost at ‘female’ rate in hybrid genetic background in presence of one dose of Lhr. In rescued hybrid males, the melanogaster-derived X chromosome appeared to complete its replication faster than autosomes. These results together have been interpreted to have suggested that Lhr suppresses the lethality of hybrids by regulating functional activities of the X chromosome(s) for dosage compensation.     

Chromosome analysis in coregonid individuals in the interspecific hybridization zone

Artificial spawning (reproduction), a popular method in fisheries, has caused the unintentional hybridization between native European whitefish (Coregonus lavaretus) and introduced peled (Coregonus peled). The resultant hybrids are fertile and their morphological identification is impossible. The study shows the genetic characteristics of the hybrid coregonid fish specimens. Analyses of the number and morphology of chromosomes were based on conventional Giemsa staining. Nucleolar Organiser Regions (NOR) were analysed using CMA₃‐ and Ag‐NOR staining. Metaphases were also investigated by C‐banding methods. Hybrid specimens had a diploid chromosome number (2n) ranging from 76 to 80 and an arm chromosome number ranging from 96 to 100. NORs were observed on the subtelo‐acrocentric and submeta‐metacentric chromosomes. Results of the present study indicate a high level of chromosomal differentiation due to rearrangement taking place in the gene pool of whitefish/peled hybrids.     

A three-locus system of interspecific incompatibility underlies male inviability in hybrids between Drosophila buzzatii and D. koepferae

In hybrids between the sibling species D. buzzatii and D. koepferae, both sexes are more or less equally viable in the F₁: However, backcross males to D. buzzatii are frequently inviable, apparently because of interspecific genetic incompatibilities that are cryptic in the F₁. We have performed a genetic dissection of the effects of the X chromosome from D. koepferae. We found only two cytological regions, termed hmi-1 and hmi-2, altogether representing 9% of the whole chromosome, which when introgressed into D. buzzatii cause inviability of hybrid males. Observation of the pattern of asynapsis of polytene chromosomes (incomplete pairing, marking introgressed material) in females and segregation analyses were the technique used to infer the X chromosome regions responsible for this hybrid male inviability. The comparison of these results with those previously obtained with the same technique for hybrid male sterility in this same species pair indicate that in the X chromosome of D. koepferae there are at least seven times more regions that produce hybrid male sterility than hybrid male inviability. We have also found that the inviability brought about by the introgression of hmi-1 is suppressed by the cointrogression of two autosomal sections from D. koepferae. Apparently, these three regions conform to a system of species-specific complementary factors involved in an X-autosome interaction that, when disrupted in backcross hybrids by recombination with the genome of its sibling D. buzzatii, brings about hybrid male inviability.     

Banding patterns of the chromosomes of Erythrocebus patas (Schreber 1774) compared to other primate species

The karyotype of Erythrocebus patas was studied using G, Q and C bands and NOR staining and compared to those of Miopithecus talapoin, Macaca mulatta and Cercopithecus aethiops. The comparison shows a large number of homoeologies, but several chromosome pairs cannot be matched. The mechanisms that may have originated the karyotypes of these species are discussed.     

Characterization of the satellite DNA Msat-160 from the species <Emphasis Type="Italic">Chionomys</Emphasis><Emphasis Type="Italic">nivalis</Emphasis> (Rodentia,Arvicolinae)

The satellite DNA Msat-160 has been previously characterized in several species of the genus Microtus. Here we present the characterization of Msat-160 from Chionomys nivalis, a species with a very primitive karyotype. As in other Microtus species analyzed, C. nivalis Msat-160 is AT rich, has a monomer length of 160 bp, is undermethylated and is mainly located in all the pericentromeric heterochromatin of all autosomes and the X chromosome, but is completely absent from the Y chromosome. Hence, our results support the hypothesis that Msat-160 was initially distributed in the pericentromeric heterochromatin of all autosomes and the X chromosome. The taxonomic status of the genus Chionomys in relation to the genus Microtus is a very interesting issue, so we constructed phylogenetic dendrograms using Msat-160 sequences from several Microtus species. Although the results were not informative about this issue, the presence of Msat-160 in C. nivalis and Microtus species suggested that both genera are closely related and that this satellite DNA was present in the common ancestor. Studies of Msat-160 in different arvicoline species could help to determine the origin of this satellite and, perhaps, to establish the phylogenetic relationships of some arvicoline groups.     

Chromosomal relationships among cultivars of Citrus reticulata Blanco, its hybrids and related species

Karyotypes of 93 individuals belonging to 18 accessions of mandarins, mandarin hybrids and two related species were analysed with the fluorochromes CMA and DAPI, to identify marker chromosomes. The karyotypes revealed highly differentiated banding patterns and could be classified in four groups (I–IV) according to the presence/absence of chromosomes with three bands (type A) or with two bands (one proximal and one terminal, type B, or both terminal, type C). The accessions of group I exhibited the simplest and homozygous karyotypes (lacking chromosome types A, B and C), represented by `Sunki' and `Cleopatra'. Group II (lacking chromosome types A and B) included three accessions of Mediterranean mandarins and `Cravo' mandarin, all of them with very similar and almost completely homozygous karyotypes. All other karyotypes of groups II and III (lacking chromosome type A) were heterozygous for one or more chromosome pairs and most of them seemed to be hybrid derivatives from non-mandarin accessions. Group IV (with chromosome types A and B) was represented only by two heterozygous hybrids (`Murcott' and `King'). The karyotype of most hybrids agrees with one of the possible combinations resulting from chromosome types segregation from their putative ancestor karyotypes, but at least `Orlando' seemed to be a more complex hybrid. Comparing with banding patterns of other Citrus species, those of group I and the Mediterranean mandarins (group II) are the best candidates to represent C. reticulata (sensu Swingle) as a true species.