Chromosome evolution in Australian rodents

The chromosome complements of 188 specimens of 29 species of Australian murid rodents belonging to the subfamilies Pseudomyinae and Hydromyinae and the Uromys/Melomys group have been compared. At least one specimen of 18 different species was successfully C-banded. — The autosomal complements of many (9) diverse Pseudomyinae, one species of Melomys and one Hydromyinae proved to be identical, comprising 48 elements in the diploid set, the two smallest autosomal pairs of which are metacentric. No other karyotype is common to more than one species. From this we conclude that these three groups have been derived from a common ancestor which also possessed such a karyotype. The genus Zyzomys is exceptional since it possesses only 44 elements and lacks the two smallest metacentrics. — Karyotypic evolution within this apparently single phyletic line has been remarkably conservative, only three rearrangements being required to derive the most divergent karyotype. Moreover most of the observed rearrangements involve pericentric inversions and only one example of a fusion was found. Considerable differences in heterochromatin content, as determined by C-banding, occur between species however. Two species proved exceptional in this respect, namely Notomys cervinus and Uromys caudimaculatus. N. cervinus possesses numerous heterochromatic short arms. In U. caudimaculatus, there is a striking difference between northern and southern populations; in the former heterochromatin is present principally in the telomeric areas of the conventional A-chromosomes whereas in the latter it is found as separate supernumerary chromosomes. — In contrast to the autosomes, the X and Y chromosomes show high inter- and intra-specific variability in both size and morphology. All of this variability can be explained in terms of variation in heterochromatin content. Moreover the amount of heterochromatin in the X and Y chromosomes is highly correlated both within and between species. The Y chromosome of Uromys caudimaculatus is, however, distinctive in that it lacks C-banding.     

SUPPRESSION OF SEX-RATIO MEIOTIC DRIVE AND THE MAINTENANCE OF Y-CHROMOSOME POLYMORPHISM IN DROSOPHILA

Like several other species of Drosophila, D. quinaria is polymorphic for X-chromosome meiotic drive; matings involving males that carry a “sex-ratio” X chromosome (X^SR) result in the production of strongly female-biased offspring sex ratios (Jaenike 1996). A survey of isofemale lines of D. quinaria from several populations reveals that there is genetic variation for partial suppression of this meiotic drive. Crossing experiments show that there is Y-linked, and probably autosomal, variation for suppression of drive. Y-linked suppressors of X-chromosome drive have now been described in several species of Diptera. I develop a simple model for the maintenance of Y-chromosome polymorphism in species polymorphic for X-linked meiotic drive. One interesting feature of this model is that, if there is a stable Y-chromosome polymorphism, then the equilibrium frequency of the standard and sex-ratio X chromosomes is determined solely by Y-chromosome parameters, not by the fitness effects of the different X chromosomes on their carriers. This model suggests that Y-chromosome polymorphism may be easier to maintain than previously thought, and I hypothesize that karyotypic variation in Y chromosomes will be found to be associated with suppression of sex-ratio meiotic drive in other species of Drosophila.     

Fluorescence pattern of the sex-chromosomes of Melandrium dioicum,stained with quinacrine-mustard

Sex-chromosomes in a male and a female clone of M. dioicum were studied by fluorescent staining with quinacrine-mustard. A dark band was found in the middle of the long arm of one X-chromosome in the female. In the other X-chromosome no band was visible, but a slightly enhanced fluorescence was shown in the same region. In the Y-chromosome a more intense fluorescence was found on both ends of the chromosome and a slight overall difference between the arms.The relation between fluorescence pattern and heterochromatin is discussed.     

Chromosome-banding study in Gerbillinae (Rodentia). I. Comparative analysis of Gerbillus poecilops, G. henleyi and G. nanus

An analysis was made of the R- and C-banding of chromosomes of the following Gerbillus species (Rodentia, Gerbillidae): G. poecilops (Taif, Saudi Arabia), G. henleyi (Oursi, Burkina Faso; Kairouan, Tunisia), and G. nanus (Taif and Hofuf, Saudi Arabia; Sind desert, Pakistan). Their karyotypes show the same chromosome number (2n = 52), but a different N.F.a. (number of autosomal arms), ranging from 58 to 62 in the species overall, and from 58 to 60 among G. nanus forms, due to pericentric inversions involving chromosomes 1 and 4. Moreover, large inter-individual variations of heterochromatin segments of chromosomes 25, X, and Y were found. Considering morphology and karyotype, G. poecilops and G. henleyi appear to be rather monomorphous species, whereas G. nanus produces a series of forms as a consequence of speciation, in which both chromosomal and morphological differentiations occur that do not correlate.     

Satellite DNA probes of Alstroemeria longistaminea (Alstroemeriaceae) paint the heterochromatin and the B chromosome,reveal a G-like banding pattern,and point to a strong structural karyotype conservation

Alstroemeria species present a well-conserved and asymmetric karyotype. The genus is divided into a Chilean clade, rich in heterochromatin, and a Brazilian clade, poor in heterochromatin. We investigated the distribution of the main repetitive sequences in the chromosomes of the Brazilian species A. longistaminea (2n = 16 + 0-6B) aiming to evaluate the role played by these sequences on the structural organization of the karyotype. In situ hybridization of the three most abundant retrotransposons, corresponding to ~ 45% of the genome, was uniformly distributed. Three satellite DNA sequences, representing near half of the whole satellite fraction (1.93% of the genome), were mainly concentrated on the heterochromatin and one of them painted the whole B chromosome. Noteworthy, some satellites were located on euchromatin, either dispersed or concentrated in clusters along the chromosomes, revealing a G-band-like pattern. The two satellites that presented more C-band- and G-band-like labeling were also hybridized in situ in two other Alstroemeria species. They revealed astonishing similar patterns of distribution, indicating an unusually structural karyotype conservation among Brazilian species.

     

Cytogenetic characterization of the ant Trachymyrmex fuscus Emery, 1934 (Formicidae: Myrmicinae: Attini) with the description of a chromosomal polymorphism

The genus Trachymyrmex is a key group in the tribe Attini because of its close phylogenetic relationship to leaf-cutter ants, Acromyrmex and Atta. Cytogenetic data are only available for five taxa of Trachymyrmex, with chromosome numbers of 2n = 12, 18, 20 and 22, and morphology with predominantly metacentric chromosomes. The aim of the present study was to characterize the karyotype of the ant Trachymyrmex fuscus Emery, 1934, by means of the number and morphology of its chromosomes, heterochromatin pattern, CMA₃ and DAPI fluorochromes in the population of two nests collected at Paraopeba, state of Minas Gerais, Brazil. Nineteen females presented 2n = 18 chromosomes (16m + 2sm) and a single male presented n = 9 (8m + 1sm). A size chromosomal polymorphism involving the short arm of the submetacentric pair was confirmed by statistical analysis, with three character conditions: heterozygous SB (with a difference in size between the short arms), standard SS (smaller short arms) and homozygote BB (bigger short arms). In the first nest, both SB and SS workers were observed. The other nest contained heterozygous (SB), homozygous (BB), and a male carrying the B chromosome (larger size). The presence of heterochromatin on all centromeric and pericentromeric chromosomes of T. fuscus suggests that the size difference observed in the submetacentric pair in the SB and BB workers is not related to the heterochromatin but to a duplication of euchromatic regions through intra- or inter- chromosomal rearrangements. The fluorochrome CMA₃ matched the C-banding markings, indicating that the heterochromatin is rich in GC base pairs. As far as we know, this is the first chromosomal polymorphism reported in the tribe Attini.     

A study of karyotypes and heterochromatic associations in Artemia,with special reference to two N. Greek populations

The chromosomes of three stains of Artemia were studied, a bisexual diploid (A. franciscana, strain GSL), and two parthenogenetic tetraploids from Northern Greece. Routine karyotypes were constructed for the diploid and tetraploid forms, as well as a Giemsa C-banding karyotype of the bisexual form; some data are also presented on the chromosomal C-banding patterns of the parthenogenetic populations. The possible significance of various chromosomal configurations that have been observed, and their relationships to heterochromatin, are discussed.This paper is dedicated to Professor C. Barigozzi for introducing us to Artemia research.Supported by a grant from the Greek Ministry of Agriculture, Request offprints from: Th. Abatzopoulos, Lab. of General Biology, School of Sciences, Aristotelian University, Thessaloniki, 54006, Greece.     

Multiple nucleolus organizer regions in Leptodactylus mystacinus (Amphibia, Anura) and comments on its systematic position in the L. fuscus group based on cytogenetic and molecular analyses

Specimens of Leptodactylus mystacinus from Brazil were karyotyped with conventional and differential staining. The 2n = 22 karyotype is similar to that found for the majority of the Leptodactylus, the karyotypic conservatism also confirmed by the similarity of the replication banding patterns with those previously described. L. mystacinus has a small amount of C-banded heterochromatin, located mainly at the centromeres, although telomeric or interstitial bands have also been noticed. With DA/CMA₃ some chromosome regions showed slightly bright fluorescence, and with DA/DAPI, no particular AT-rich repetitive region was observed. Silver staining showed an extensive inter- and intraindividual variation in the number and position of Ag-positive regions, in 1p, 4p, 8p, 8q, and 11p. Nevertheless, FISH using rDNA probes confirmed only the signals on the short arms of chromosomes 4 and 8 as true NORs. The remaining silver stained regions are probably due to the heterochromatin with some affinity to the Ag-staining. Phylogenetic analysis based on partial cytochrome b sequence revealed that L. mystacinus forms a basal branch, so that the presence of multiple NORs in pairs 4 and 8 in this species indicates an autapomorphy. Supported by FAPESP and CNPq.     

Cytophotometric DNA determinations and autoradiographic studies in salivary gland nuclei from larvae with different karyotypes in Drosophila melanogaster

Cytophotometric DNA determinations in Feulgen stained mitotic diploid chromosome sets of neuroblasts from larvae of Drosophila melanogaster stocks, which possess different karyotypes, show significant differences between the 4C values, caused by an additional or deficient X- and Y-chromosome depending on the karyotype. The ranges of polytenic DNA size classes are theoretically expected to be doublings of the corresponding 4C mean value of each karyotype. The extinction integral data of nuclei with completely duplicated 4C quantities exclusively fall into the range of the expected size classes. Not all data falling into the range of a size class necessarily originate from duplicated nuclei, because the limits of the DNA size classes cannot be determined by measurements, but must be estimated from the confidence limits of the corresponding 4C mean value. The validity of the mitotic 4C values of the karyotypes X/X and X/Y is tested using data from non-labeled interphase nuclei, where extinction integral data accumulate in two groups. The larger values (= G₂-nuclei) confirm the 4C values of mitotic chromosome sets, and the lower values (= G₁-nuclei) are just half of these. Extinction integrals from individual, ³H-thymidine non-incorporating polytene salivary gland nuclei accumulate in distinct, non-overlapping groups which are always complete doublings of the preceding smaller group. In each karyotype, the most frequent data of each group are in accord with the 4C doublings. The data from labeled nuclei alternate with those from unlabeled nuclei. The measured DNA values of individual polytene nuclei that did not incorporate any ³H-thymidine, demonstrate that all chromosomal DNA replicates completely during polytenization of the chromosomes in the larval salivary gland nuclei of Drosophila melanogaster. Specifically, this would mean that the heterochromatic Y-chromosome replicates as well as the partially heterochromatic X-chromosome along with the autosomes. There is no indication of underreplicating heterochromatin.     

Cytogenetics Status of Four Ctenomys Species in the South of Brazil

This report is a review of 20 years of cytogenetics studies in tuco-tucos from the south of Brazil and the implications for the evolution of these species as well as the relation of these data with the geological history of the Coastal Plain of southern Brazil. Two forms of Ctenomys torquatus, 2n = 44 and 46, are separated by a geographic barrier. Ctenomys flamarioni shows a constant karyotype (2n = 48) and presents high variability in FN due to constitutive heterochromatin variation. Ctenomys minutus presents the highest chromosomic variation among the species in the south of Brazil (2n = 42–50) with three chromosomic hybrid zones. Ctenomys lami, like C. minutus, presents a high chromosomic variation due to fusions and fissions of chromosome pairs 1 and 2. Both species present a close evolutionary relationship, including a chromosomic hybrid zone.     

Evolution of human Y-chromosome DNA

We have used human male-specific 3.4 kb Hae III restriction endonuclease fragments to explore the evolutionary history of man's Y-chromosome. We have identified four sets of reiterated, sequences on the basis of their relative sequence homology with autosomal DNA. The sequences account for approximately 40% of the human Y-chromosome, are interspersed within the same 3.4 kb Hae III fragments, are heterogeneous and contain all reiterated DNA previously demonstrated to be specific for the Y-chromosome (it-Y DNA). Y-specific 3.4 kb Hae III sequences do not reassociate with either human female or ape DNA at standard reassociation criteria. However, approximately half of it-Y DNA (cross reacting it-Y) reassociates with both human female and ape DNA at reduced reassociation criteria. The remaining half (Y-specific it-Y) retains its specificity for the human Y-chromosome. These two sets of it-Y DNA have distinct reiteration frequencies and thermal stabilities with their Y-chromosome homologs. Non-Y-specific 3.4 kb Hae III sequences reassociate with both human female and ape DNA at standard reassociation criteria. The abundance of these non-Y-specific sequences decreases as a function of their evolutionary distance from man. One subset of non-Y-specific 3.4 kb Hae III sequences forms stable duplexes with human Y-chromosome DNA and with human and ape autosomal DNA. No detectable base-mismatch occurs among these homologs suggesting complete conservation of these sequences during primate evolution. The second subset of Non-Y-specific Hae III sequences form stable duplexes with human Y-chromosome DNA but highly mismatched duplexes with human and ape autosomal DNA.The finding that homologs of 3.4 kb Hae III sequences are not found within the Y-chromosome of apes but are only present in autosomes suggests that 3.4 kb Hae III sequences are largely autosomal in origin. Since autosomal homologs of most 3.4 kb Hae III-sequences exhibit a greater degree of divergence than those localized to the Y-chromosome, their evolutionary history seems to be chromosome-dependent.Our findings are not easily correlated with the comparative morphology of primate Y-chromosomes and suggest that sequence rearrangement has been a major event in the evolution of the human Y-chromosome. The significance of the specific interspersion of four sets of reiterated sequences, with distinct evolutionary histories, within a repeating unit specific to the human Y-chromosome is not clear. The apparent conservation of at least some of these reiterated sequences suggests they may be of functional importance.     

Typification and redelimitation of Nepenthes alata with notes on the N. alata group,and N. negros sp. nov. from the Philippines

In the course of studies to typify what has been regarded as the most widespread and common of the endemic Philippines species of Nepenthes, N. alata Blanco, we were able to review the morphological variation in what we previously regarded as a polymorphic species. This led us to redelimit that species in a narrower sense, to resurrect N. graciliflora Elmer, and to recognise N. negros sp. nov., here assessed as ‘Critically Endangered’ (CR) using the IUCN standard. The Nepenthes alata group is characterised and a key to its species is provided.     

A new karyotype for <Emphasis Type="Italic">Cavia magna</Emphasis> (Rodentia: Caviidae) from an estuarine island and <Emphasis Type="Italic">C. aperea</Emphasis> from adjacent mainland

We describe a new karyotype for Cavia magna Ximenez, 1980 from an estuarine island and the karyotype of Cavia aperea Erxleben, 1777 from an adjacent mainland. The species have differences in diploid number (2n), autosomal fundamental number, quantity, and distribution of heterochromatin as dissimilar distributions of the nucleolus-organizing regions (Ag-NORs). The C. aperea karyotype has a diploid number of 64 as previously reported for C. aperea and most other Cavia species. In contrast, this new C. magna karyotype exhibits a variant diploid number of 2n = 62, considering that previous work reported a karyotype of 2n = 64 for C. magna. The discovery of a distinct diploid number within C. magna represents the first record of intra-specific chromosomal variation in a species of Caviidae. The diploid number of 2n = 62, heterochromatin quantity, Ag-NOR distributions, and inversed X chromosome from this population of C. magna are as seen in the geographically proximate (Cavia intermedia Cherem Olimpio and Ximenez; intermediate Cavy). These data provide further evidence supporting C. magna as the sister species of C. intermedia.     

Banded karyotypes of mole rats (Spalax, Spalacidae, Rodentia) from Turkey: a comparative analysis

Chromosome banding (G-, C- and Ag-NOR) analysis was carried out on 27 specimens of Sphalax ehrenbergi from seven localities and two specimens of S. leucodon from one locality, all from Turkey. No chromosomal variation was detected in S. ehrenbergi populations from Elazig, Siverek, Diyarbakir and Birecik having the same diploid numbers (2n = 52) and morphology of chromosomes (NFa = 72). The karyotypes of mole rats from Tarsus and Gaziantep possessed the identical diploid number (2n = 56) but different numbers of autosomal arms: NFa = 68 in the Tarsus and NFa = 78 in the Gaziantep populations. Chromosomes of S. leucodon from Malaty (2n = 60, NFa = 74) differed distinctly in the C-banding pattern from all S. ehrenbergi cytotypes by the almost entire absence of heterochromatin in acrocentric autosomes and the presence of heterochromatin arms iin subtelocentric autosomes. Nucleolar organizing regions were found mainly on three pairs of chromosomes, but some differences in their localization were revealed. Comparison of G-banded chromosomes showed, that most chromosomes have a similar pattern. The types of chromosomal rearrangemetns were revealed due to the banding methods.     

Cytogenetic evolution in the plethodontid salamander genus Aneides

Chromosomal variation in the plethodontid salamander genus Aneides was analyzed using C-banding, to visualize heterochromatin distribution, as well as silver and fluorochrome banding, to reveal nucleolus organizer regions. Evidence is presented that Aneides has undergone pronounced karyological divergence from a Plethodon-like ancestor. It is concluded that this divergence reflects accelerated rates of chromosomal rearrangements in Aneides relative to Plethodon. These rearrangements have involved mainly pericentric inversions that have occurred in one or more of the three smallest pairs of chromosomes in the karyotype. Karyological variation within and between species of Aneides was used to analyze phylogenetic relationships within the genus. This analysis revealed at least two karyologically distinct species that had not been previously recognized by morphological criteria.     

XX:XY sex chromosome system with X heterochromatinization: an early stage of sex chromosome differentiation in the Neotropic electric eel Eigenmannia virescens

An early stage of sex chromosome differentiation is reported to occur in the electric eel Eigenmannia virescens (Pisces, Sternopygidae) from populations of two tributaries of the Paraná river system (Brazil). Cytogenetic studies carried out in the two populations showed that the Mogi-Gua?u population is characterized by 2n = 38 chromosomes and undifferentiated sex chromosomes and the Tietê population presents 2n = 38 both for males and females and an XX:XY sex chromosome system. The X-chromosome is acrocentric, easily recognized by the presence of a conspicuous heterochromatin block in its distal portion; the Y-chromosome is probably one of the medium sized acrocentrics present in the male karyotype. BrdU induced R-bands of the two populations did not reveal any difference in the euchromatic regions of the chromosomes. AluI and HaeIII restriction enzyme digestion patterns and chromomycin A3 staining of the X-chromosome are presented. The possible role of heterochromatinization in the evolution of sex chromosomes in fish is discussed.     

XY<Subscript>1</Subscript>Y<Subscript>2</Subscript> chromosome system in <Emphasis Type="Italic">Salinomys delicatus</Emphasis> (Rodentia,Cricetidae)

Salinomys delicatus is considered a rare species due to its restricted and patchy distribution, poor records and low abundances. It is also the phyllotine with the lowest known diploid chromosome number (2n = 18), however its sex chromosome system has never been described. Here, we studied the chromosomes of six females and three males with bands G, C, DAPI/CMA₃ and meiosis. In males, the chromosome number was 2n = 19, with one large metacentric X-chromosome and two medium-sized acrocentrics absent in females. The karyotype of females was the same as previously described (2n = 18, FN = 32), with X-chromosomes being metacentric and the largest elements of the complement. In males, the two acrocentrics and the large metacentric form a trivalent in meiotic prophase. This indicates that S. delicatus has XY₁Y₂ sex chromosomes, which is confirmed by G and DAPI bands. Constitutive heterochromatin (CH) is restricted to small pericentromeric blocks in all chromosomes. The X-chromosome shows the largest block of centromeric CH, which could favor the establishment of this X-autosome translocation. This sex chromosome system is rare in mammals and, compared with other phyllotine rodents, S. delicatus seems to have undergone a major chromosome restructuring during its karyotypic evolution.     

Comparative Analysis (Hippotragini versus Caprini, Bovidae) of X-Chromosome's Constitutive Heterochromatin by in situ Restriction Endonuclease Digestion: X-Chromosome Constitutive Heterochromatin Evolution

The Bovidae X-chromosome shows a considerable variation, in contrast to the preservative autosomal conservatism. The X-chromosome variation is mostly a consequence of the constitutive heterochromatin (CH) variation; in what respect to its amount and position. This is especially common among the non-Bovinae subfamilies and tribes. In order to characterize the X-chromosome CH in non-Bovinae species--Hippotragini and Caprini tribes--we have used restriction endonuclease digestion on fixed chromosomes and sequential C-banding. With these techniques we were able to distinguish between the two X-chromosome types (Hippotragini and Caprini) CH, in what respect to its position and molecular nature. Moreover, we define at least, six subclasses of CH in both X-chromosome types analyzed. Evolutionary considerations were draw based on the results obtained. The technology used here for the analysis of the Bovidae X-chromosome CH showed to be more evolutionary informative than the classical approaches.     

Chromosomal evolution and phylogenetic analyses in <Emphasis Type="Italic">Tayassu pecari</Emphasis> and <Emphasis Type="Italic">Pecari tajacu</Emphasis> (Tayassuidae): tales from constitutive heterochromatin

The mammalian family Tayassuidae (peccaries) is confined to the New World and comprises three recognized extant species, white-lipped (Tayassu pecari), collared (Pecari tajacu) and chacoan (Catagonus wagneri) peccaries, which exhibit distinct morphological and chromosomal features. The phylogenetic relationships among the tayassuids are unclear and have instigated debate over the palaeontological, cytogenetic and molecular aspects. Constitutive heterochromatin analysis can be used in understanding the phylogenetic relationships between related species. Here we describe, for the first time, the constitutive heterochromatin (C-positive heterochromatin) of two tayassuid species, Tayassu pecari and Pecari tajacu. We demonstrate that in situ restriction endonuclease digestion with sequential C-banding could be a complementary tool in the study of constitutive heterochromatin heterogeneity in chromosomes of the Tayassuidae. Our characterization of peccary chromosomes suggests that the Pecari tajacu autosomal karyotype is more primitive and has accumulated great diversity in its constitutive heterochromatin. This idea is supported by several other studies that analysed nuclear and mitochondrial sequences of the living peccary species. Finally, the tayassuid X chromosome primitive form seems to be the one of Tayassu pecari.     

Heterochromatin variation and spermatogenesis in Nesokia

A probable role of heterochromatin variation in male meiosis has been evaluated using fertile and infertile Indian mole rat males (Nesokia) with polymorphic X and/or Y chromosomes. A comprehensive study of tubular histology, meiotic progression, and X-Y chromosome pairing was undertaken. Despite heterochromatin variation, spermatogenesis was found to be complete in all individuals. Patterns of X-Y synaptonemal complex pairing varied considerably from extensive synapsis in individuals with a normal heterochromatin complement, through end-to-end synapsis, to X and Y univalents in those with different degrees of loss of heterochromatin. Changes in the gonadal histology corresponding to heterochromatin variation were also observed. Loss of some coding DNA sequences in polymorphic X-chromosomes otherwise located at specific sites in the X-chromosome heterochromatin have been linked directly to modifications of the reproductive process. This is thought to be mediated by an altered X-chromosome activity during spermatogenesis or regulation of other locus/loci involved in fertility or reproduction.