Molecular cytogenetics and characterization of a ZZ/ZW sex chromosome system in Triportheus nematurus (Characiformes, Characidae)

Chromosomes of Triportheus nematurus, a fish species from family Characidae, were analyzed in order to establish the conventional karyotype, location of C-band positive heterochromatin, Ag-NORs, GC- and AT-rich sites, and mapping of 18S and 5S rDNA with fluorescence in situ hybridization (FISH). The diploid number found was 2n = 52 chromosomes in both males and females. However, the females presented a pair of differentiated heteromorphic chromosomes, characterizing a ZZ/ZW sex chromosome system. The Z chromosome was metacentric and the largest one in the karyotype, bearing C-positive heterochromatin at pericentromeric and telomeric regions. The W chromosome was middle-sized submetacentric, appearing mostly heterochromatic after C-banding and presenting heterogeneous heterochromatin composed of GC- and AT-rich regions revealed by fluorochrome staining. Ag-NORs were also GC-rich and surrounded by heterochromatic regions, being located at the secondary constriction on the short arms of the second chromosome pair, in agreement with 18S rDNA sites detected with FISH. The 18S and 5S rDNA were aligned in tandem, representing an uncommon situation in fishes. The results obtained reinforce the basal condition of the ZZ/ZW sex system in the genus Triportheus, probably arisen prior to speciation in the group.     

Karyotype differentiation between two stickleback species (Gasterosteidae)

The stickleback family (Gasterosteidae) of fish is less than 40 million years old, yet stickleback species have diverged in both diploid chromosome number (2n) and morphology. We used comparative fluorescence in situ hybridization (FISH) on 2 stickleback species, Gasterosteus aculeatus (2n = 42) and Apeltes quadracus (2n = 46), to ascertain the types of chromosome rearrangements that differentiate these species. The A. quadracus karyotype contains more acrocentric and telocentric chromosomes than the G. aculeatus karyotype. By using bacterial artificial chromosome probes from G. aculeatus in our FISH screen, we found that 6 pericentric inversions and 2 chromosome fusions/fissions are responsible for the greater number of acrocentric and telocentric chromosomes in A. quadracus. While most populations of G. aculeatus have an XX/XY sex chromosome system, A. quadracus has a ZZ/ZW sex chromosome system, as previously reported. However, we discovered that a population of A. quadracus from Connecticut lacks heteromorphic sex chromosomes, providing evidence for unexpected sex chromosome diversity in this species.     

Differentiated ZZ/ZW sex chromosomes in Apareiodon ibitiensis (Teleostei,Parodontidae): cytotaxonomy and biogeography

Conventional and molecular chromosomal analyses were carried out on three populations of Apareiodon ibitiensis sampled from the hydrographic basins of the São Francisco River and Upper Paraná River (Brazil). The results reveal a conserved diploid number (2n = 54 chromosomes), a karyotype formula consisting of 50 m‐sm + 4st and a ZZ/ZW sex chromosome system that has not been previously identified for the species. C‐banding analysis with propidium iodide staining revealed centromeric and terminal bands located in the chromosomes of the specimens from the three populations and allowed the identification of heteromorphism of heterochromatin regions in the Z and W chromosomes. The number of 18S sites located through fluorescent in situ hybridization (FISH) varied between the populations of the São Francisco and Upper Paraná Rivers. The location of 5S rDNA sites proved comparable in one pair of metacentric chromosomes. Thus, the present study proposes a ZZ/ZW sex chromosome system for A. ibitiensis among the Parodontidae, and a hypothesis is presented regarding possible W chromosome differentiation stages in this species through DNA accumulation, showing geographical variations for this characteristic, possibly as a consequence of geographical reproductive isolation.     

A Novel ZZ/ZW Sex Chromosome System for the Genus Leporinus (Pisces, Anostomidae, Characiformes)

A wide range of sex chromosome mechanisms, including simple and multiple chromosome systems is characteristic of fishes. The Leporinus genus represent a good model to study sex chromosome mechanisms, because an unambiguous ZZ/ZW sex chromosome system was previously described for seven species, while the remaining studied species of the genus do not show differentiated sex chromosomes. The occurrence of sex chromosomes in Leporinus trifasciatus and Leporinus sp2 from the Araguaia river, Amazon basin, Brazil, was here investigated. ZZ/ZW sex chromosomes were detected for both species. The Z and W chromosome morphology of L. trifasciatus is the same as described for other species of the genus Leporinus. However, the Z and W chromosomes of L. sp2 were quite different in their morphology and banding pattern suggesting that the ZW system of this species have originated independently from the ZW system previously described for other Leporinus.     

Chromosome evolution within theOrnithogalum tenuifolium complex (Hyacinthaceae), with special emphasis on the evolution of bimodal karyotypes

Hypotheses on the evolution of the karyotypes of 8 chromosome races (2n = 4, 6, 8, 10, 12, 16-two forms, 26) within theOrnithogalum tenuifolium complex are discussed. Four of the karyotypes are strictly bimodal: 2n = 8 (6 long and two short chromosomes), 2n = 10 (6 long and 4 short chromosomes), 2n = 12 (6 long and 6 short chromosomes) and 2n = 16 (12 long and 4 short chromosomes). The hypotheses are tested by means of measurements of nuclear DNA content, studies of meiosis and pollen fertility of hybrids, and comparisons of karyotype morphology. The results indicate that the E. African 2n = 12 chromosome race is the most primitive and has given rise to the other chromosome races. The 2n = 6 race is found to have a significantly higher fitness than the 2n = 12 race.     

ZZ/ZW sex chromosome system in an undescribed species of the genus Apareiodon (Characiformes, Parodontidae)

The chromosomes of an undescribed species of the genus Apareiodon (Characiformes, Parodontidae) from the Verde River, a headwater affluent of the Tibagi River (Paraná State, Brazil), were investigated using conventional Giemsa and Ag stainings, C-banding, CMA(3) fluorescence and fluorescent in situ hybridization (FISH) using 18S and 5S rDNA probes. The diploid chromosome number was 2n = 54, with the karyotype composed of 48 meta/submetacentric and six subtelocentric chromosomes in males, and 47 meta/submetacentric + seven subtelocentric chromosomes in females. The difference is hypothesized to be due to a ZZ/ZW heteromorphic sex chromosome system, a cytotaxonomic characteristic previously observed only in some species of the genus Parodon (family Parodontidae). The presence of similar and/or identical heteromorphic sex chromosome systems might suggest that species of the genera Parodon and Apareiodon bearing ZZ/ZW heteromorphic sex chromosomes likely constitute a monophyletic group, a hypothesis to be tested by a robust phylogeny of the family.     

AMPHIPLOID ORIGIN OF TWO ENDEMIC RACES OF ASTER (ASTERACEAE) IN SOUTHERN CALIFORNIA

Two cytological races of Aster which morphologically resemble A. ascendens but differ from it in chromosome number are found in the Transverse and Peninsular ranges of southern California. One of these races is relatively common, can usually be separated morphologically from A. ascendens, and has a somatic chromosome number of 36; it corresponds to the endemic species A. bernardinus. The other race is rare and localized, is morphologically almost indistinguishable from A. ascendens, and has a somatic chromosome number of 42. The chromosome numbers and karyotypes of the two races suggest that they originated through backcrosses of A. ascendens (x = 13) to its respective parent taxa, A. falcatus (x = 5) and A. occidentalis (x = 8).     

Differentiation of repetitive DNA sites and sex chromosome systems reveal closely related group in Parodontidae (Actinopterygii: Characiformes)

Parodon and Apareiodon lack sufficiently consistent morphological traits to be considered a monophyletic group in Parodontidae. Species within this family are either sex-homomorphic or sex-heteromorphic (i.e., lacking a differentiated sex chromosome system, ZZ/ZW or ZZ/ZW(1)W(2)). In this study, a DNA fragment from the heterochromatin segment of the W chromosome of Apareiodon ibitiensis (named WAp) was microdissected and used for in situ mapping of nine Parodontidae species. The species were also characterized using a satellite DNA probe (pPh2004). The species were phylogenetically clustered according to 17 characters, which were examined by both classical and molecular cytogenetic techniques. Given the present results, the single ZZ/ZW sex chromosome system seems to have been derived from a paracentric inversion of a terminal WAp site onto the proximal regions of the short arms of a metacentric chromosome pair, followed by WAp site amplification. We reason that these events restrained recombination and favored differentiation of the W chromosome in some species. Moreover, co-hybridization experiments targeting the WAp and pPh2004 repetitive DNA sites of A. affinis suggest that the ZZ/ZW(1)W(2) sex chromosomes of this species may have arisen from a translocation between the proto-sex chromosome and an autosome. Our phylogenetic analysis corroborates the hypothesis of sex chromosome differentiation and establishes groups of closely related species. The phylogenetic reorganization in response to these new data supports the presence of internal monophyletic groups within Parodontidae.     

A cytological study ofPelargonium sect.Eumorpha (Geraniaceae)

The chromosome numbers of seven species ofPelargonium sect.Eumorpha have been determined from material of known wild origin, and karyotypic comparisons have been made. Within the section there is variation in basic chromosome number (x = 4, 8, 9, 11), variation in chromosome size, and two species have polyploid races. The three species with chromosome numbers based on x = 11 have the smallest chromosomes (1.0–1.5 µm); chromosomes are larger (1.0–3.0 µm) in the other species.P. elongatum has the lowest chromosome number in the genus (2n = 8).P. alchemilloides is exceptional in that it has four cytotypes, 2n = 16, 18, 34 and 36, and the form with 2n = 36 has large chromosomes (2.0–5.0 µm). Evidence from a synthesized hybrid suggests thatP. alchemilloides with 2n = 16 may be of polyploid origin. The three species based on x = 11 appear to be more closely related to species from other sections ofPelargonium that have the same basic chromosome number and small chromosome size, rather than to other species of sect.Eumorpha.     

Population cytogenetics of the genus Caledia (Orthoptera: Acridinae)

The genus Caledia contains two species. C. species nova 1 is restricted to the Oriomo Plateau of S.W. Papua and has a complement of twelve telocentric chromosomes. The second species C. captiva has a much wider distribution pattern—from S.W. Papua in the North, down the entire Eastern seaboard of Australia to Southern Victoria. It is also found in the Northern Territory. Although the chromosome number is the same as C. species nova 1, four major and distinct chromosomal races can be distinguished in C. captiva. — The basic ancestral race is found in Tropical North Queensland at the base of the Cape York Peninsula. All twelve chromosomes are telocentric and the karyotypic organization is similar to that found in C. species nova 1 and in other Acridines. A second, general purpose karyotypic race has a wide distribution between S.W. Papua, Arnhem Land and the East Australian coast as far South as Brisbane. It is considered a derivative form of the ancestral type and is fixed for small pericentric inversions on seven pairs of chromosomes. In the South-Eastern Queensland region there exists a further race which carries large pericentric inversions on all the autosomes and the X chromosome. The situation here is confounded since the basic chromosomes can be represented as either acro or telocentrics. Various levels of polymorphism for the inversions exist between different chromosomes in different populations indicating considerable differentiation within this zone. This race is almost completely surrounded by the general purpose karyotype where the races are contiguous in certain parts of the range. — The South-Eastern corner of Australia is characterised by a chromosome race quite different from those found further North. Here a complex pericentric inversion system exists involving a series of seven small inversions and larger inversions on chromosomes 1, 2, 4 and 10. Chromosomes 2 and 4, in particular, are highly polymorphic. — The presence and persistence of these 4 chromosomal races can be accounted for in terms of the known climatic changes which have occurred in this region in the recent past.     

A new chromosomal race (2n=44) of Nannospalax xanthodon from Turkey (Mammalia: Rodentia)

A new karyotype for blind mole rats was recorded in Tunceli province in Eastern Turkey. The karyotype contained 44 chromosomes, including 13 biarmed pairs, 7 acrocentric pairs, and one heteromorphic pair with a submetacentric and an acrocentric homologue in the autosomal complement (FNa=69). The X chromosome was submetacentric and the Y chromosome medium-sized subtelocentric (FN=73). Distinct dark centromeric C-bands were observed on most of the biarmed and three pairs of the acrocentric autosomes. The NORs were detected on short arms of three subtelocentric pairs and one acrocentric pair of autosomes. The diploid number of chromosomes and the karyotype characteristics observed are obviously unique among hitherto studied populations of blind mole rats and the complement can be evaluated as a new chromosome race of Nannospalax xanthodon. The distribution ranges of individual chromosome races of the species recorded in Eastern Anatolia are revised and possible interracial hybridization is discussed in respect of the finding of a new race.     

Sex chromosome system ZZ/ZW in Apareiodon hasemani Eigenmann, 1916 (Characiformes,Parodontidae) and a derived chromosomal region

Parodontidae fish show few morphological characteristics for the identification of their representatives and chromosomal analyses have provided reliable features for determining the interrelationships in this family. In this study, the chromosomes of Apareiodon hasemani from the São Francisco River basin, Brazil, were analyzed and showed a karyotype with 2n = 54 meta/submetacentric chromosomes, and a ZZ/ZW sex chromosome system. The study revealed active NORs located on pair 11 and additional 18S rDNA sites on pairs 7 and 22. The 5S rDNA locus was found in pair 14. It showed a pericentric inversion regarding the ancestral condition. The satellite DNA pPh2004 was absent in the chromosomes of A. hasemani, a shared condition with most members of Apareiodon. The WAp probe was able to detect the amplification region of the W chromosome, corroborating the common origin of the system within Parodontidae. These chromosomal data corroborate an origin for the ZW system of Parodontidae and aid in the understanding of the differentiation of sex chromosome systems in Neotropical fishes.     

Chromosomal evolution in the lizard genus Varanus (reptilia).

The karyotypes have been determined of 16 of the 32 species of the genus Varanus, including animals from Africa, Israel, Malaya and Australia. A constant chromosome number of 2n = 40 was observed. The karyotype is divided into eight pairs of large chromosomes and 12 paris of microchromosomes. A series of chromosomal rearrangements have become established in both size groups of the karyotype and are restricted to centromers shifts, probably caused by pericentric inversion. Species could be placed in one of six distinct karyotype groups which are differentiated by these rearrangements and whose grouping does not always correspond with the current taxonomy. An unusual sex chromosome system of the ZZ/ZW type was present in a number of the species examined. The evolutionary significance of these chromosomal rearrangements, their origin and their mode of establishment are discussed and related to the current taxonomic groupings. The most likely phylogenetic model based on chromosome morphology, fossil evidence and the current distribution of the genus Varanus is presented.     

Karyotypic diversification due to Robertsonian rearrangements in Phyllodactylus lanei Smith, 1935 (Squamata, Gekkonidae) from Mexico

We analyzed chromosomes of male and female individuals of Phyllodactylus lanei Smith, 1935 (Squamata, Gekkonidae) from Chamela-Cuixmala Biosphere Reserve, Jalisco state, Mexico. The karyotype constructed for these specimens is composed of 19 pairs of telocentric chromosomes (2n = 38, FN = 38). This karyotype, due to Robertsonian fusions/fissions, differs from the one previously reported in samples from the State of Guerrero, which probably belonged to a different subspecies (2n = 33−34, FN = 40−41). Moreover, a presumed ZW sex chromosome system was not confirmed in the presently studied individuals.      

Chromosome polymorphism and C-banding variation of the brachypterous grasshopper Podisma sapporensis Shir. (Orthoptera, Acrididae) in Hokkaido, northern Japan

The grasshopper Podisma sapporensis consists of two main chromosome races in Hokkaido. The western group of populations of P. sapporensis, belonging to the XO race, has a diploid number of chromosomes 2n = 23 in the male and 2n = 24 in the female (sex determination XO male/XX female). The eastern group of populations of this species, belonging to the XY race, differs from the western one as a result of Robertsonian translocation between the originally acrocentric X chromosome and M5 autosome in homozygous state, having resulted in the forming of chromosome sex determination neo-XY male/neo-XX female (2n = 22). These races are geographically isolated by the mountainous system consisting of the Mts Daisetsu and Hidaka range, occupying the central part of the island. The hybrid zones between the races have not so far been discovered. Various levels of polymorphism for the pericentric inversions and C-banding variation exist in different chromosomes throughout populations in both chromosome races. In some solitary populations (the population at the summit of Mt Yotei, populations in the vicinity of Naganuma, Oketo, and Tanno) pericentric inversions are fixed in some pairs of chromosomes, which enables marking of the discrete karyomorphes. In the Mt Daisengen population all chromosomes are two-armed as a result of fixing the pericentric inversions. These facts contradict karyotypical conservatism of the tribe Podismini. The level of diversity of P. sapporensis karyotypes could provide a new perspective on the evolutionary process of different karyotype in Orthoptera. The considerable occurrence of polymorphism in chromosomes suggests that karyotypic diversification is undergoing in P. sapporensis. The authors also proposed that P. sapporensis would be divided into four chromosome subraces in the XO chromosome race and two chromosome subraces in the XY race, on the basis of karyotypic features. These races may have been established by fundamental climatic changes during the glacial epoch.     

Sex chromosome differences among the three races (alpha, beta, gamma) of the snail intermediate host of Schistosoma mekongi, Neotricula aperta

Chromosome analyses were performed on three races (alpha, beta, gamma) of Neotricula aperta which were previously reported to show variation in the chromosome number and pairing in meiosis. We used an air-drying method, which was more reliable for chromosome preparations from fresh animal tissues than a squash method. Each of the races had the same number of chromosomes, 2n=33 for males, and 2n=34 for females. The sex determination system was XO type (male: 32+X, female: 32+XX). The sex chromosome of each race was distinct in its morphology, but not in its length. Karyotyping revealed that the X-chromosome of the alpha race was metacentric (M), whereas it was acrocentric (A) in the beta and the gamma races. An association between the X-chromosome and a small autosome in meiosis suggested that a neo-Y chromosome probably lies in the terminal region of the small autosome.     

The cytology of Brachycome lineariloba

A comparison of the karyotypes of races D (2n=8), E (2n=10), B (2n=12) and C (2n=16) of B. lineariloba suggests that these races have in common a basic set of four chromosome pairs, and that the higher chromosome number races are related to race D by successive chromosome addition. — A study of meiosis in B × C and A₁ × B hybrids supports this contention and elucidates the homologies of the additional chromosomes. — Meiotic pairing in hybrids between A and C is very complex. At present it can only be stated that there are extensive interchromosomal homologies between the two races. — Two phyletic schemes of the relationships of the races are considered. The second, which is favoured, involves successive chromosome addition, with the quasidiploid race E (2n = 10) giving rise to race B by diploidisation of the univalent chromosomes. This scheme is supported by features of univalent behaviour in the various races and their hybrids. — The ecogeographic distribution pattern of the races shows replacement of D by E by B by C as the species extends into more arid and more harsh environments. This replacement is also associated with increasing vigour. — It seems most likely that the addition chromosomes are derived from a race A (2n=4) source since they are added always by twos, and each addition increases both vigour and drought tolerance. Race A is the most vigorous and one of the most drought tolerant of the five races.It is suggested that the evolution of the races can be related to the increasing aridity of the Late Pleistocene and Recent geological epochs.     

Karyological differentiation and speciation in C. MediterraneanAnacamptis (Orchidaceae)

Anacamptis pyramidalis is a variable and wide-spread European-Mediterranean taxon. Beside a dominant cytotype with 2n = 36 it includes cytotypes with 2n = 54 and 63 in northern Tuscany (and the Eastern Pyrenees) and one with 2n = 72 on Malta. In contrast,A. urvilleana, formerly often misidentified and included inA. pyramidalis, is a monomorphic and distinct species, endemic to the Maltese Islands. It has 2n = 36, can be clearly separated by morphological and anatomical features and is isolated from partly sympatric populations ofA. pyramidalis with 2n = 72 by differences in chromosome number, flowering time and habitat preference.     

Chromosome evolution in fish: sex chromosome variability in Eigenmannia virescens (Gymnotiformes: Sternopygidae)

New data are presented on the sex chromosomes of the fish species Eigenmannia virescens (Gymnotiformes, Sternopygidae). A new finding, involving the occurrence of ZZ/ZW sex chromosomes, is described in specimens sampled from the S?o Francisco and Amazon river basins in Brazil. All individuals had a chromosome number of 2n = 38. The homologs of the sex chromosome pair from the S?o Francisco river basin sample differed only in their morphology, while those from the Amazonian sample differed both in morphology and heterochromatin pattern. A possible model for the evolution of the sex chromosomes in E. virescens is proposed, including data from populations from the Paraná (Brazil) river basin, in which male heterogamety has already been described. The occurrence of different sex chromosome systems in species and populations of the neotropical freshwater fish fauna is discussed.