Multiple sex chromosome system of X1X1X2X2/X1X2)Y type in lutjanid fish, Lutjanus quinquelineatus (Perciformes)

The karyotype and other chromosomal markers as revealed by C-banding and Ag-staining were studied in Lutjanus quinquelineatus and L. kasmira (Lutjanidae, Perciformes). While in latter species, the karyotype was invariably composed of 48 acrocentric chromosomes in both sexes, in L. quinquelineatus the female karyotype had exclusively 48 acrocentric chromosomes (2n = 48) but that of the male consisted of one large metacentric and 46 acrocentric chromosomes (2n = 47). The chromosomes in the first meiotic division in males showed 22 bivalents and one trivalent, which was formed by an end-to-end association and a chiasmatic association. Multiple sex chromosome system of X₁X₁X₂X₂/X₁X₂Y type resulting from single Robertsonian fusion between the original Y chromosome and an autosome was hypothesized to produce neo-Y sex chromosome. The multiple sex chromosome system of L. quinquelineatus appears to be at the early stage of the differentiation. The positive C-banded heterochromatin was situated exclusively in centromeric regions of all chromosomes in both species. Similarly, nucleolus organizer region sites were identified in the pericentromeric region of one middle-sized pair of chromosomes in both species. The cellular DNA contents were the same (3.3 pg) between the sexes and among this species and related species.     

A multiple sex-chromosome system in Antarctic ice-fishes

Summary We have studied the chromosomes of 11 of the 15 known species of the notothenioid family Channichthyidae, the specialized whiteblooded Teleosts endemic to the Southern Ocean (ice-fishes). In the female sex, all studied species have the same diploid number of forty eight mostly acrocentric (uniarmed) chromosomes; however there is an interspecific variability in the chromosome morphology, type and quantity of repetitious DNA (usually seen as heterochromatin) localization of silver-stained nucleolar organizers. At least five of the studied species show a multiple sex-chromosome system possibly originated by the translocation of an autosome on an early Y gonosome morphologically similar to the X: the digametic males (2n = 47) show a X₁Y X₂ and the homogametic females (2n = 48) a X₁X₁X₂X₂ gonosomic constitution. This peculiar sex determining mechanism, otherwise rare in Teleosts, can be considered apomorphic in the same way as other morphofunctional characters usually interpreted as adaptive in these fishes.Some of the data presented here were collected during the European Polarstern Study (EPOS) sponsored by the European Science Foundation     

Chromosome banding in amphibia

A cytogenetic study performed on a population of the South American leptodactylid frog Eleutherodactylus maussi revealed multiple sex chromosomes of the X₁X₁X₂X₂/X₁X₂Y (=XXAA/XXA^Y) type. The diploid chromosome number is 2n=36 in all females and 2n=35 in most males. The multiple sex chromosomes originated by a centric fusion between the original Y chromosome and a large autosome. In male meiosis the X₁X₂Y (=XXA^Y) multiple sex chromosomes form a classical trivalent configuration. E. maussi is the first species discovered in the class Amphibia that is distinguished by a system of multiple sex chromosomes. Only one single male was found in the population with 2n=36 chromosomes and lacking the Y-autosomal fusion. This karyotype (XYAA) is interpreted as the ancestral condition, preceding the occurrence of the Y-autosome fusion.by H.C. Macgregor     

Cytogenetic characterization of ten araneomorph spiders (Araneae): Karyotypes and meiotic features

Karyotypes, sex chromosome systems and meiotic characteristics are reported for ten spider species belonging to the families Gnaphosidae, Philodromidae, Salticidae, Oxyopidae and Sicariidae by using standard Giemsa staining. The male diploid numbers (2n) and sex chromosome systems are as follows: Berinda hakani 2n = 22 (X₁X₂), Berinda ensigera 2n = 22 (X₁X₂), Trachyzelotes lyonneti 2n = 22 (X₁X₂), Trachyzelotes malkini 2n = 22 (X₁X₂), Zelotes caucasius 2n = 22 (X₁X₂) (Gnaphosidae); Thanatus pictus 2n = 28 (X₁ X₂), Tibellus macellus 2n = 24 (X₁ X₂) (Philodromidae); Neon reticulatus 2n = 21 (X₀) (Salticidae); Peucetia virescens 2n = 28 (X₁X₂) (Oxyopidae) and Loxosceles rufescens 2n = 21 (X₁ X₂Y) (Sicariidae). All species have monoarmed chromosomes with the exception of L. rufescens that has biarmed (metacentric and submetacentric) chromosomes. The obtained data are the first results for the genera Berinda, Trachyzelotes and Neon. Additionally, with the exception of L. rufescens, all species are being chromosomally analyzed for the first time.     

Mechanisms of chromosomal evolution and its possible relation to natural history characteristics in Ancistrus catfishes (Siluriformes: Loricariidae)

Ancistrus is the most speciose genus of the tribe Ancistrini, with 58 valid species and many yet to be described. Cytogenetic studies were conducted on five apparently undescribed species from the Amazon basin, which showed different diploid numbers: Ancistrus sp. Purus (2n = 34); Ancistrus sp. Macoari (2n = 46); Ancistrus sp. Dimona (2n = 52); Ancistrus sp. Vermelho (2n = 42) and Ancistrus sp. Trombetas (2n = 38). All species possessed only one pair of NOR‐carrying chromosomes, but with extensive variation in both the location on the chromosome as well as in the position of the ribosomal sites on the karyotype. The karyotypic evolution of Ancistrus species seems to be based on chromosomal rearrangements, with a tendency to a reduction of the diploid number. Two new instances of XX/XY sex chromosomes for Ancistrus species, based on the heteromorphism in the male karyotype, were also recorded. The large karyotypic diversity among Ancistrus species may be related to biological and behavioural characteristics of these fish that include microhabitat preferences, territoriality and specialized reproductive tactics. These characteristics may lead to a fast rate of fixation of chromosomal mutations and eventually speciation across the basin.     

The Point Evaluation Method for Approximating Function Means,Variances and Covariances

A perennial problem in statistics is the determination of biases, variances and covariances for functions of random variables X₁, X₂, …, X_n which themselves have a known distribution. A common approach is through equations based upon Taylor series approximations but a “point evaluation” method may sometimes be a useful alternative. This involves approximating the multivariate distribution of the X variables by the 2ⁿ points given by X₁=μ₁±₁, X₂ = μ₂ ±₂, …, X_n = = μ_n μ_n, where μ_i is the mean and σ_i the standard deviation of Xi, with appropriate point weights. An advantage over the Taylor series approach is that function derivatives do not have to be explicitely calculated. The point evaluation method is particularly useful in cases where the X variables are uncorrelated. Then the evaluation of the 2ⁿ points can be replaced by the evaluation of 2n points. The point evaluation method is illustrated with powers of a normally distributed variable, and with estimation of gene frequencies from ABO blood group frequencies.     

Complex sex-linked fusion heterozygosity in the Australian huntsman spider Delena cancerides (Araneae: Sparassidae)

In the vast majority of spider species studied to date, the karyotype is homogeneous in morphology and exclusively telocentric. The sex-determining system consists of one to three X chromosomes in the male and, correspondingly, two to six in the female. This is the case in species of huntsman spiders belonging to the genera Heteropoda (2n=40+3X), Isopoda, Olios, and Pediana (2n=40+3X) and some populations of the colonial species Delena cancerides (2n=40+3X). In other populations of D. cancerides, wholesale fusion of the karyotype has occurred, reducing the standard huntsman karyotype of 43 telocentric chromosomes to 21 metacentrics and 1 telocentric. Eight of the centric fusion products, including an X-autosome fusion, are maintained in the heterozygous condition in males and, with the single telocentric, form a chain of nine chromosomes at meiosis. The two complexes comprising the chain behave as neo-X and neo-Y chromosomes, and thus the ancestral X₁X₂X₃X₁X₁X₂X₂X₃X₃ sex-determining system has been converted to a system of six X and four Y chromosomes in the male and twelve X chromosomes in the female. Since sex-linked complex heterozygosity is also found in a number of species of social termites, it is suggested that such heterozygosity may have adaptive significance for a colonial lifestyle. Breakdown products of the chain of nine are present in specimens of D. cancerides from Canberra and these appear to represent hybrid products between the 2n=22 and 2n=43 forms. Hybridisation may also have been involved in the origin of the chain-forming races.     

Chromosomal diversification in populations of Characidium cf. gomesi (Teleostei, Crenuchidae)

Comparative cytogenetic studies carried out in two populations of Characidium cf. gomesi from Botucatu region, SP, Brazil, showed a similar karyotypic structure in a diploid number of 50 chromosomes, 32 metacentric and 18 submetacentric chromosomes for males and 31 metacentric and 19 submetacentric chromosomes for females as well as a ZZ-ZW sex chromosome system. Differences between both populations, however, were found in relation to the occurrence of B chromosomes and the distribution of 18S and 5S ribosomal DNA (rDNA) sites. Characidium cf. gomesi from the Alambari Stream, a component of the Tietê River basin, revealed 18S rDNA on Z and W chromosomes, while this gene was located on autosomes in the sample from the Paranapanema River basin. The 5S rDNA sites were observed in a single chromosomal pair (number 25) in the populations from Paranapanema and in two pairs in the specimens from Tietê (numbers 20 and 25). Besides that, in the sample from Paranapanema, both inter and intra-individual variations were found due to the occurrence of up to four heterochromatic supernumerary chromosomes in the cells. The life mode of this fish, restricted to headwaters and subjected to frequent breakdown into sub-populations, may have contributed to the fixation of such chromosomal differences. The karyotypic similarities found in the analysed populations, however, suggest that all are descended from the same ancestor group whereas their differences indicate that they are already existing in reproductively isolated populations.     

Banded karyotype of spined loach Cobitis taenia and triploid Cobitis from Poland

The present work provides new data on the banding pattern of diploid Cobitis taenia and its triploid hybrid females, which belong to the diploid–polyploid complex in the Vistula River tributary. C-banding, silver-staining (Ag), and fluorescent staining with chromomycin A₃ techniques were used to describe the diploid and triploid karyotype. The karyotype of Cobitis taenia of 2n=48 was characterised by one pair of NOR-bearing subtelocentric chromosomes and at least four chromosomes with CMA₃-positive sites. The C-positive heterochromatin was present in the centromeres of almost all chromosomes and the pericentromeric regions of several metacentric and submetacentric chromosomes. The triploid females of 3n=74 had two pairs of chromosomes with active NORs. The NORs-sites were located terminally on two biarmed and two uniarmed chromosomes. The CMA₃-staining revealed at least six A₃-positive sites. The C-banded and A₃-stained triploid karyotype was composed of haploid set of Cobitis taenia and diploid set of unidentified species, so heterochromatin pattern confirmed the possibility of their hybrid origin. The characteristics of banded diploid and triploid karyotype, and the hypothetical karyotype of an unknown species of 2n=50 is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

A complex sex-chromosome system in the hare-wallaby Lagorchestes conspicillatus Gould

Among specimens of the spectacled hare-wallaby Lagorchestes conspicillatus Gould (Marsupialia, family Macropodidae) 4 males had 15 chromosomes and 2 females 16 chromosomes. The sex chromosomes are X₁X₁X₂X₂ in the female and X₁X₂Y in the male, the Y being metacentric and both X chromosomes are acrocentric. In about 96% of sperm mother cells at meiosis the sex chromosomes form a chain trivalent and in more than 99% of these this orients convergently so that the X₁ and X₂ move to the same pole. Evidence is presented that L. conspicillatus has evolved from a form with 22 chromosomes including a small X and a minute Y. Autoradiographic studies show that the proximal fifth of the X₁ chromosome replicates late. This is probably the ancestral X chromosome which has been translocated to an autosome. The fate of the original Y is obscure but an hypothesis is proposed that it forms the centromeric region of the Y. A single male had 14 chromosomes and was heterozygous for a translocation involving the centric fusion of two acrocentric autosomes. In about 30% of sperm mother cells the autosomal trivalent did not disjoin regularly but, despite this, all secondary spermatocytes observed at metaphase 2 had balanced complements of chromosomes. It is assumed that unbalanced secondary spermatocytes died before reaching metaphase.     

The chromosomes of Schoutedenia lutea (homoptera,aphidoidea, greenideinae), with an account of meiosis in the male

Somatic chromosomes of both sexes and chromosome behaviour during spermatogenesis were studied in the aphid Schoutedenia lutea (van der Goot). Four long but unequal chromosomes in females were interpreted as X chromosomes (X₁X₁X₂X₂) with one member of an autosome pair attached to one X₁, and the other member to one X₂, so that the four long chromosomes were actually X₁+A, X₁, X₂+A, X₂. Males (normally XO in aphids) received X chromosomes corresponding in relative length to the two longest (X₁+A, X₂+A) in females. During spermatogenesis parallel pairing occurred in prophase 1 and the X₁ and X₂ chromosomes became associated via their autosomal segments. In anaphase I, the autosomal segment became detached from one of the X chromosomes and entered the non-viable (non-X-bearing) spermatocyte, while the viable spermatocyte received both X₁ and X₂ (either one of which still carried an autosome) and the haploid set of free autosomes. The consequences for sex determination and zygote formation of this unusual system are discussed; a stable chromosomal constitution for the zygote can be achieved only at the expense of considerable gamete wastage.     

Chromosome spreading of associated transposable elements and ribosomal DNA in the fish Erythrinus erythrinus. Implications for genome change and karyoevolution in fish

Background  

The fish, Erythrinus erythrinus, shows an interpopulation diversity, with four karyomorphs differing by chromosomal number, chromosomal morphology and heteromorphic sex chromosomes. Karyomorph A has a diploid number of 2n = 54 and does not have differentiated sex chromosomes. Karyomorph D has 2n = 52 chromosomes in females and 2n = 51 in males, and it is most likely derived from karyomorph A by the differentiation of a multiple X₁X₂Y sex chromosome system. In this study, we analyzed karyomorphs A and D by means of cytogenetic approaches to evaluate their evolutionary relationship.     

Chromosomal rearrangements and the first indication of an ♀X1X1X2X2/♂X1X2Y sex chromosome system in Rineloricaria fishes (Teleostei: Siluriformes)

Rineloricaria is the most diverse genus within the freshwater fish subfamily Loricariinae, and it is widely distributed in the Neotropical region. Despite limited cytogenetic data, records from southern and south-eastern Brazil suggest a high rate of chromosomal rearrangements in this genus, mirrored in remarkable inter- and intraspecific karyotype variability. In the present work, we investigated the karyotype features of Rineloricaria teffeana, an endemic representative from northern Brazil, using both conventional and molecular cytogenetic techniques. We revealed different diploid chromosome numbers (2n) between sexes (33♂/34♀), which suggests the presence of an ♀X₁X₁X₂X₂/♂X₁X₂Y multiple sex chromosome system. The male-limited Y chromosome was the largest and the only biarmed element in the karyotype, implying Y-autosome fusion as the most probable mechanism behind its origination. C-banding revealed low amounts of constitutive heterochromatin, mostly confined to the (peri)centromeric regions of most chromosomes (including the X₂ and the Y) but also occupying the distal regions of a few chromosomal pairs. The chromosomal localization of the 18S ribosomal DNA (rDNA) clusters revealed a single site on chromosome pair 4, which was adjacent to the 5S rDNA cluster. Additional 5S rDNA loci were present on the autosome pair 8, X₁ chromosome, and in the presumed fusion point on the Y chromosome. The probe for telomeric repeat motif (TTAGGG)_n revealed signals of variable intensities at the ends of all chromosomes except for the Y chromosome, where no detectable signals were evidenced. Male-to-female comparative genomic hybridization revealed no sex-specific or sex-biased repetitive DNA accumulations, suggesting a presumably low level of neo-Y chromosome differentiation. We provide evidence that rDNA sites might have played a role in the formation of this putative multiple sex chromosome system and that chromosome fusions originate through different mechanisms among different Rineloricaria species.     

Cytogenetic,hybridization and molecular evidence of four cytological forms of Anopheles nigerrimus (Hyrcanus Group) in Thailand and Cambodia

Thirteen isoline colonies of Anopheles nigerrimus were established from individual wild‐caught females collected from cow‐baited traps at locations in Thailand and Cambodia. Three types of X (X₁, X₂, X₃) and 4 types of Y (Y₁, Y₂, Y₃, Y₄) chromosomes were recovered, according to differing amounts of extra heterochromatin. Four karyotypic forms were designed depending upon apparently distinct figures of X and Y chromosomes, i.e., Form A (X₁, X₂, X₃, Y₁), B (X₂, X₃, Y₂), C (X₁, Y₃), and D (X₃, Y₄). Forms C and D were new metaphase karyotypes discovered in this study. Form A appeared to be common in both Thailand and Cambodia. Forms B and D were found to be rather specific to southern and northeastern Thailand, respectively, whereas Form C was confined to Cambodia. Hybridization experiments among the eight isoline colonies, which were representative of four karyotypic forms of An. nigerrimus, demonstrated genetic compatibility in giving viable progenies and synaptic salivary gland polytene chromosomes through F₂‐generations. These results elucidated the conspecific relationship, comprising four cytological forms within this taxon. Supportive evidence was confirmed further by very low intraspecific sequence variations (average genetic distance = 0.002–0.007) of the nucleotide sequences in ribosomal DNA [second internal transcribed spacer (ITS2)] and mitochondrial DNA [cytochrome c oxidase subunit I (COI) and subunit II (COII)].     

SYNTHETIC HYBRIDS OF NEW WORLD AND OLD WORLD AGROPYRONS: III. AGROPYRON REPENS × TETRAPLOID AGROPYRON SPICATUM

Three hybrids of A. repens, 2n = 42, × A. spicatum, 2n = 28, and two reciprocal hybrids were obtained from emasculated and unemasculated crosses, respectively. The 35-chromosome hybrids tended to be morphologically intermediate between the parent species but resembled A. repens more closely than A. spicatum. A. repens behaved cytologically as a segmental autoallohexaploid, and A. spicatum acted cytologically as an autotetraploid. Mean chromosome associations of 8.04 I, 12.72 II, 0.41 III, 0.06 IV, and 0.009 V were observed in 116 hybrid cells at metaphase I. Most chromosome pairing in the hybrids was attributed to autosyndesis. A. spicatum, A. repens, and their hybrids were represented by genome formulas of SSSS, R₁R₁X₁X₁X₂X₂, and SSR₁X₁X₂, respectively. Hybrid fertility ranged from 0.02 to 0.69 seeds per spikelet.     

Barley Chromosome Identification Using Genomic in Situ Hybridization in the Genome of Backcrossed Progeny of Barley–Wheat Amphiploids [Hordeum geniculatum all. (2n = 28) × Triticum aestivum L. (2n = 42)] (2n = 70)

Genomic in situ hybridization (GISH) has been used to study characteristics of the formation of alloplasmic lines detected among self-pollinated backcrossed progeny (BC₁F₅–BC₁F₈) of barley–wheat amphiploids [Hordeum geniculatum All. (2n = 28) × Triticum aestivum L. (2n = 42)] (2n = 70). The chromosome material of the wild barley H. geniculatum has been shown to contribute to these lines. For example, fifth-generation plants (BC₁F₅) had genotypes (2n= 42w + 2g), (2n = 42w + 1g + 1tg), and (2n = 41w + 1g), where w is common wheat chromosomes, g is barley (H. geniculatum) chromosomes, and tg is the telocentric chromosome of wild barley. Beginning from theBC₁F₆ generation, alloplasmic telocentric addition lines (2n= 42 + 2tg) and (2n = 42 + 1tg) appear. This lines has been found cytogenetically unstable. The progeny of each of these cytological types include not only the (2n= 42 + 2tg) and (2n = 42 + 1tg) addition plants, but also plants with the monosomic (2n = 41 + 1tg) and the disomic (2n = 40 + 2tg) substitutions, as well as the (2n = 41 + 2tg) plants, which lack one wheat chromosome and have two telocentric barley chromosomes. It has been demonstrated that the selection for well-filled grains favors the segregation of telocentric addition lines (2n = 42 + 2tg) and (2n = 42 + 1tg).     

Karyological studies on the Indian spiders IX. Chromosome constitution in two cribellate species

The haploid number of chromosomes inStegodyphus sarasinorum (Eresidae) andUloborus plumipes (Uloboridae) is thirteen (11+X₁X₂) and ten (8+X₁X₂) respectively. The sex-chromosomes are of equal size in the former and unequal in the latter. All chromosomes are acrocentric in both species.     

Whole chromosome painting reveals independent origin of sex chromosomes in closely related forms of a fish species

The wolf fish Hoplias malabaricus includes well differentiated sex systems (XY and X₁X₂Y in karyomorphs B and D, respectively), a nascent XY pair (karyomorph C) and not recognized sex chromosomes (karyomorph A). We performed the evolutionary analysis of these sex chromosomes, using two X chromosome-specific probes derived by microdissection from the XY and X₁X₂Y sex systems. A putative-sex pair in karyomorph A was identified, from which the differentiated XY system was evolved, as well as the clearly evolutionary relationship between the nascent XY system and the origin of the multiple X₁X₂Y chromosomes. The lack of recognizable signals on the sex chromosomes after the reciprocal cross-FISH experiments highlighted that they evolved independently from non-homologous autosomal pairs. It is noteworthy that these distinct pathways occur inside the same nominal species, thus exposing the high plasticity of sex chromosome evolution in lower vertebrates. Possible mechanisms underlying this sex determination liability are also discussed.     

Chromosome polymorphism in <Emphasis Type="Italic">Rineloricaria pentamaculata</Emphasis> (Loricariidae,Siluriformes) of the Paraná River basin

Cytogenetic analysis in three Rineloricaria pentamaculata populations revealed diploid number 2n = 56 chromosomes, karyotype formula 8m/sm + 48st/a and FN = 64. Owing to the presence of the heteromorphic chromosome pair with a big submetacentric chromosome and a small acrocentric one in both males and females, 42.9% of specimens in the Tauá Stream population had the karyotype formula 9m/sm + 47st/a and FN = 65. Analysis of the nucleolus-organizing region by Ag-NOR and FISH techniques showed a single NOR system at pair 5 for R. pentamaculata populations of the Keller River and the Tauá Stream. However, specimens of populations of the Tatupeba Stream had multiple NOR systems at pairs 5 and 8. A constitutive heterochromatin pattern in R. pentamaculata is mainly distributed in the pericentromeric and telomeric regions with interstitial markers in certain chromosomes. Heterochromatin is located in the telomeric and centromeric positions of the acrocentric chromosome in the heteromorphic pair of the Tauá Stream population. In the submetacentric chromosome the markings are located in the telomeric (short arm), pericentomeric and interstitial (long arm) positions. The origins of polymorphisms are discussed.