1971 Chen A comparative chromosome study of twenty killifish species of the genus Fundulus (Teleostei: Cyprinodontidae) Abstract Female karyotypes from ovarian cell cultures of 20 species of killifish (Fundulus) ranged in diploid number from 32 to 48, but in arm number (NF) from 48 to 52. The “small” F chromosomes, which constituted the fundamental elements in the karyotype, were evenly graded in length. The “large” biarmed chromosomes (L), which were about twice the length of the average Fs, characterized only those species with 2N less than 48 chromosomes. And among these species, an increase in complement by a pair of L's was always accompanied by a decrease of two pairs of A's, indicating Robertsonian changes by the centric fusion of two A's to form one L chromosome. Other diagnostic chromosome characters included: the number and structure of biarmed and satellited F chromosomes and the percentage of F's with relatively short short-arms (SSA). Besides centric fusion, mechanisms of chromosomal evolution in Fundulus probably included pericentric inversion, producing biarmed F chromosomes from acrocentric F's and partial loss of a chromosome segment producing smaller biarmed F chromosomes from larger ones. The percentage of SSA chromosomes generally decreases from relatively primitive to specialized species. The presumably most primitive species have only SSA type acrocentric F chromosomes. The 20 Fundulus species were classified into 2 major groups according to the percentage of SSA chromosomes: the SSA group, including 3 subgroups, had more than 50% SSA's; the LSA group, including 2 subgroups, had fewer than 50% SSA's. This classification based only on karyotypic characters generally agreed with others based on gross morphological characters. A possible evolutionary scheme is proposed to account for the derived killifish karyotypes. 1977 Kligerman Rapid Chromosome Preparations from Solid Tissues of Fishes ABSTRACT A technique is described for obtaining well-spread metaphases from solid tissues of fishes without the use of methodologies that rely on tissue grinders, centrifuges, digestive enzymes, or tissue culture. This procedure involves the formation of a cell suspension from acetic alcohol fixed tissues using 50% acetic acid. The suspension is applied to a warm (50 °C) slide using a micropipette.Solid tissue preparations may be stained by any of the conventional dyes or treated to reveal Q-bands, C-bands, and nucleolar organizers. Large numbers of slides offish chromosomes can be made easily and rapidly using this procedure 1978 Khuda Bukhsh Chromosomes in three species of fishes, Aplocheilus panchax (Cyprinodontidae), Lates calcerifer (Percidae) and Gadusia chapra (Clupeidae). About 800 species of fishes have so far been cytologically investigated (MANNA and KHUDA-BUKHSH 1977) as against some 20,000 taxonomically known ones (DENTON 1973). Extension of the cytological data is, therefore, a necessity. The present communication deals with the diploid number and karyomorpological analysis of somatic chromosomes in both sexes of Aplocheilus panchax, in male Lates calcerifer and germinal chromosomes in male Aplocheilus panchax and Gadusia chapra. 1985 Ewolonu A Multiple Sex Chromosome System in the Annual Killifish, Nothobranchius guentheri Abstract Since the discovery of multiple sex chromosomes in fishes by Uyeno & Miller (1971) similar systems have been described in a number of fish species. Such a system is described here for Nothobranchius guentheri (Pfeffer) along with some taxonomic clarification regarding species name in this group. 1991 Elder Chromosomal divergence and heterogameity in two annual killifishes of the genus Pterolebias ABSTRACT The annual killifish (Cyprinodontidae) species Pterolebias hoignei and Pterolebias zonatus from the Orinoco basin, Venezuela, have been regarded as conspecific ecomorphs, but P. hoignei has a diploid number of 46, whereas P. zonatus has a diploid number of 42. Metacentric chromosome counts also differ between these two species. Pterolebias hoignei has a sexually dimorphic karyotype, while P. zonatus does not. Although the diploid number is the same for both sexes, male P. hoignei possess a single large Y chromosome not present in the female karyotype. The hypothesis of conspecificity is rejected on the basis of karyotypic divergence found between the species. Key words: Cyprinodontidae, karyotype divergence, sexual dimorphism. 1992 Magtoon Distribution and cytogenetics of killifish, aplocheilus panchax (Hamilton-Buchman) from the central, Thailand [1992] Purpose of this experiment was to study on distribution and cytogenetics study of Aplocheilus panchax. Ten specimens of A. panchax 2.5-3.5 mm in SL, were used. An observation of mitotic metaphase chromosome prepared from gill, intestine and kidney tissue by the Uwa and Ojima's method (1981) were made. Levan et al.,'s method of chromosome classification was employed. The findings were as follows: 1) Apocheilus panchax were widely distributed in the Mae Nam Choa Phraya Basin. 2) The diploid chromosome number of A. panchax was 40. The Karyotype consisted of 4 paires of metacentric, 2 pairs of submetacentric, and 14 pairs of acrocentric chromosomes. The arm number was 52. These results will be useful for cytotaxonmy of fishes. 1993 Elder Karyotypes of nine Venezuelan annual killifishes (Cyprinodontidae), with comments on karyotype The degree of chromosomal variation reported here and in the literature appears not to be as high among the New World as among the Old World annuals. The difference in karyotypic variability of these two groups, having similar life histories and reproductive strategies, suggests that habitat stochasticity, associated with annual reproductive strategies, may not have been the primary force mediating chromosomal differentiation in both of these ... 1995 Brosset Evolution as a Lottery Conflicting with Evolution via Sexual Selection in African Rain Forest—Dwelling Killifishes (Cyprinodontidae, Rivulinae, (Diapteron) Populations of cyprinodont fishes (killifish) of the subfamily Rivulinae in the Afrotropical and Neotropical regions presumably split about 100 million years ago, when Gondwanaland fragmented and Africa began to be isolated. Among the species-rich fauna of African rivulines, no extant species can cope with salinity and none is found closer than 500–1000 m to the sea. Although some American mud skipper-like rivulines such as Guianese representatives of the genus Anableps (A. anableps, A. microlepis) live in estuaries, cyprinodonts have never become fully marine, so their presence in both south America and Africa is a measure of their antiquity (Kingdon, 1990). On both continents, freshwater cyprinodonts present many similarities including sexual dimorphism, bright male coloration, elaborate displays, and population dynamics, as for example in species of Aphyosemion in west central Africa, and of the genus Rivulus in French Guiana (Brosset, unpublished). Moreover, annual as well as nonannual species of killifish have evolved in both regions (Simpson, 1979). Whether these similarities are the result of common ancestry from a tropical Gondwanian stock or of striking evolutionary convergence is nonetheless questionable. What is certain is that both lineages have been prone to local speciation. The extensive radiations of African killifishes are the focus of the present chapter 2005 Coughlin Chromosome evolution in eukaryotes: a multi-kingdom perspective In eukaryotes, chromosomal rearrangements, such as inversions, translocations and duplications, are common and range from part of a gene to hundreds of genes. Lineage-specific patterns are also seen: translocations are rare in dipteran flies, and angiosperm genomes seem prone to polyploidization. In most eukaryotes, there is a strong association between rearrangement breakpoints and repeat sequences. Current data suggest that some repeats promoted rearrangements via non-allelic homologous recombination, for others the association might not be causal but reflects the instability of particular genomic regions. Rearrangement polymorphisms in eukaryotes are correlated with phenotypic differences, so are thought to confer varying fitness in different habitats. Some seem to be under positive selection because they either trap favorable allele combinations together or alter the expression of nearby genes. There is little evidence that chromosomal rearrangements cause speciation, but they probably intensify reproductive isolation between species that have formed by another route. 2007 Schubert Chromosome evolution by Ingo Schubert Abstract The idea of evolution as a principle for the origin of biodiversity fits all phenomena of life, including the carriers of nuclear inheritance, the chromosomes. Insights into the evolutionary mechanisms that contribute to the shape, size, composition, number and redundancy of chromosomes elucidate the high plasticity of nuclear genomes at the chromosomal level, and the potential for genome modification in the course of breeding processes. Aspects of chromosome fusion, as exemplified by karyotype evolution of relatives of Arabidopsis, have recently received special attention. 2011 Ellegren Sex-chromosome evolution: recent progress and the influence of male and female heterogamety Hans Ellegren Abstract | It is now clear that sex chromosomes differ from autosomes in many aspects of genome biology, such as organization, gene content and gene expression. Moreover, sex linkage has numerous evolutionary genetic implications. Here, I provide a coherent overview of sex-chromosome evolution and function based on recent data. Heteromorphic sex chromosomes are almost as widespread across the animal and plant kingdoms as sexual reproduction itself and an accumulating body of genetic data reveals interesting similarities, as well as dissimilarities, between organisms with XY or ZW sex-determination systems. Therefore, I discuss how patterns and processes associated with sex linkage in male- and female-heterogametic systems offer a useful contrast in the study of sex-chromosome evolution. 2011 Paes da Cruz Karyotype description and evidence of multiple sex chromosome system X1X1X2X2/X1X2Y in Potamotrygon aff. motoro and P. falkneri (Chondrichthyes: Potamotrygonidae) in the upper Paraná River basin, Brazil Cytogenetic analysis of Potamotrygon aff. motoro and P. falkneri indicated the occurrence of an X1X1X2X2/X1X2 Y multiple sex chromosome system in both species, with 2n = 66 chromosomes for females and 2n = 65 chromosomes for males. The nucleolus organizer regions (NORs) identified using Ag-NOR technique showed that both species have multiple Ag-NORs (5 to 7 chromosomes stained). C-banding technique indicated the presence of heterochromatic blocks in the centromeric regions of almost all chromosomes in both species. Through this study there was evidence of heterogeneity in the karyotypes, which suggests that chromosomal rearrangements such as inversions and/or translocations occurred during the chromosomal evolution in two species of this genus. Key words: Cytogenetic, NOR, C-band, Fish, Stingrays. 2011 Polacik Asymmetric Reproductive Isolation between Two Sympatric Annual Killifish with Extremely Short Lifespans Abstract Background> Interspecific reproductive isolation is typically achieved by a combination of intrinsic and extrinsic barriers. Behavioural isolating barriers between sympatric, closely related species are often of primary importance and frequently aided by extrinsic factors causing spatial and temporal interspecific separation. Study systems with a severely limited role of extrinsic factors on reproductive isolation may provide valuable insights into how reproductive isolation between sympatric species is maintained. We used no-choice experimental set-up to study reproductive barriers between two closely related sympatric African killifish species, Nothobranchius furzeri and Nothobranchius orthonotus. These fish live in small temporary savannah pools and have complete spatial and temporal overlap in reproductive activities and share a similar ecology. Principal Findings We found that the two species display largely incomplete and asymmetric reproductive isolation. Mating between N. furzeri males and N. orthonotus females was absent under standard experimental conditions and eggs were not viable when fish were forced to mate in a modified experimental setup. In contrast, male N. orthonotus indiscriminately mated with N. furzerifemales, the eggs were viable, and offspring successfully hatched. Most spawnings, however, were achieved by male coercion and egg production and embryo survival were low. Behavioural asymmetry was likely facilitated by mating coercion from larger males of N. orthonotus and at relatively low cost to females. Interestingly, the direction of asymmetry was positively associated with asymmetry in post-mating reproductive barriers. Significance We showed that, in fish species with a promiscuous mating system and multiple matings each day, selection for strong mate preferences was relaxed. This effect was likely due to the small proportion of resources allocated to each single mating and the high potential cost to females from mating refusal. We highlight and discuss the fact that males of rarer species may often coercively mate with females of a related, more abundant species. 2012 Berdan LINKING ECOLOGICAL DIVERGENCE AND REPRODUCTIVE ISOLATION: A TALE OF TWO KILLIFISH Gene flow and recombination directly oppose genetic differentiation and speciation because they homogenize allele combinations that are unique to each population/species (Felsenstein 1981; Rieseberg 2001; Ortiz-Barrientos, Reiland et al. 2002; Butlin 2005). Under divergence with gene flow, either very strong selection or some other mechanism that reduces recombination rates is needed to maintain different allelic combinations. 2012 Betran Why Chromosome Palindromes? Abstract We look at sex-limited chromosome (Y or W) evolution with particular emphasis on the importance of palindromes. Y chromosome palindromes consist of inverted duplicates that allow for local recombination in an otherwise nonrecombining chromosome. Since palindromes enable intrachromosomal gene conversion that can help eliminate deleterious mutations, they are often highlighted as mechanisms to protect against Y degeneration. However, the adaptive significance of recombination resides in its ability to decouple the evolutionary fates of linked mutations, leading to both a decrease in degeneration rate and an increase in adaptation rate. Our paper emphasizes the latter, that palindromes may exist to accelerate adaptation by increasing the potential targets and fixation rates of incoming beneficial mutations. This hypothesis helps reconcile two enigmatic features of the “palindromes as protectors” view: (1) genes that are not located in palindromes have been retained under purifying selection for tens of millions of years, and (2) under models that only consider deleterious mutations, gene conversion benefits duplicate gene maintenance but not initial fixation. We conclude by looking at ways to test the hypothesis that palindromes enhance the rate of adaptive evolution of Y-linked genes and whether this effect can be extended to palindromes on other chromosomes.

2012 Liu Chromosome Evolution and Genome Miniaturization in Minifish Background Paedocypris is a newly established genus of fish in Southeast Asia. Paedocypris is characterized by several unique features, including a tiny adult size (thus named miniature fish or minifish), fragmentary habitats of acidic peat blackwater swamps, an unusual reproduction mode and truncated development. These peculiarities lend themselves excellent for studying chromosome evolution and rapid speciation in vertebrates but also make them highly controversial for the phylogenetic position. Methodology and Principal Findings We have established an organ procedure to prepare chromosome spreads from tiny organs of minifish and performed a cytogenetic study on two species of the genus Paedocypris, namely P. carbunculus (Pc) and P. sp. “Singkep” (Ps). We found 30 and 34 chromosomes in diploid cells of Pc and Ps, respectively, which are unusual in teleost fishes. The diploid metaphase has 5 pairs of metacentrics and 7 pairs of subtelocentrics in Pc compared to 3 pairs of metacentrics and 11 pairs of subtelocentrics in Ps, whereas the haploid metaphase contains 5 metacentrics and 7 subtelocentrics in Pc compared to 3 metacentrics and 11 subtelocentrics Ps. Chromosome behavior in first meiosis revealed the presence of a chromosomal ring consisting of 2 metacentrics in Pc, suggesting that centric fusion rather than fission was responsible for the karyotypic evolution from Ps to Pc. Flow cytometry revealed that Pc had a 45% nuclear staining intensity relative to medaka whose genome is 700 Mb in size and contains 0.81 pg DNA. The Pc genome should have 315 Mb in length and 0.36 pg of DNA, which represent one of the smallest values in vertebrates, suggesting genome miniaturization in this organism. Conclusions Our data demonstrate that gross chromosome rearrangements and genome miniaturization have accompanied the evolution of Paedocypris fishes. Our data also place Paedocypris outside currently described taxa of the Cypriniformes. 2013 Bachtrog Y-chromosome evolution: emerging insights into processes of Y-chromosome degeneration Abstract | The human Y chromosome is intriguing not only because it harbours the master-switch gene that determines gender but also because of its unusual evolutionary history. The Y chromosome evolved from an autosome, and its evolution has been characterized by massive gene decay. Recent whole-genome and transcriptome analyses of Y chromosomes in humans and other primates, in Drosophila species and in plants have shed light on the current gene content of the Y chromosome, its origins and its long-term fate. Furthermore, comparative analysis of young and old Y chromosomes has given further insights into the evolutionary and molecular forces triggering Y-chromosome degeneration and into the evolutionary destiny of the Y chromosome. 2014 Arezo Sex determination in annual fishes: searching for the master sex-determining gene in Austrolebias charrua (Cyprinodontiformes, Rivulidae) ABSTRACT Evolution of sex determination and differentiation in fishes involves a broad range of sex strategies (hermaphroditism, gonochorism, unisexuality, environmental and genetic sex determination). Annual fishes inhabit temporary ponds that dry out during the dry season when adults die. The embryos exhibit an atypical developmental pattern and remain buried in the bottom mud until the next rainy season. To elucidate genomic factors involved in the sex determination in annual fish, we explored the presence of a candidate sex-specific gene related to the cascade network in Austrolebias charrua. All phylogenetic analyses showed a high posterior probability of occurrence for a clade integrated by nuclear sequences (aprox. 900 bp) from both adults (male and female), with partial cDNA fragments of A. charrua from juveniles (male) and the dsx D. melanogaster gene. The expressed fragment was detected from blastula to adulthood stages showing a sexually dimorphic expression pattern. The isolated cDNA sequence is clearly related to dsx D. melanogaster gene and might be located near the top of the sex determination cascade in this species. 2014 Ezaz Repetitive Sequence and Sex Chromosome Evolution in Vertebrates Abstract Sex chromosomes are the most dynamic entity in any genome having unique morphology, gene content, and evolution. They have evolved multiple times and independently throughout vertebrate evolution. One of the major genomic changes that pertain to sex chromosomes involves the amplification of common repeats. It is hypothesized that such amplification of repeats facilitates the suppression of recombination, leading to the evolution of heteromorphic sex chromosomes through genetic degradation of Y or W chromosomes. Although contrasting evidence is available, it is clear that amplification of simple repetitive sequences played a major role in the evolution of Y and W chromosomes in vertebrates. In this review, we present a brief overview of the repetitive DNA classes that accumulated during sex chromosome evolution, mainly focusing on vertebrates, and discuss their possible role and potential function in this process. 2014 Silva do Nascimento Karyotype Patterns of Hypsolebias antenori (Cyprinodontiformes: Rivulidae): An Endangered Killifish of the Semiarid Region of Brazil Abstract Annual fish which belong to the order Cyprinodontiformes constitute an excellent model for evolutionary studies. their short life cycle, distribution in ecologically dynamic environments, and low agility make them favorable for genetic analyses. The species Hypsolebias antenori (Rivulidae), encountered in seasonal pools located in the semiarid region of Northeastern Brazil, has been the object of surveys with a view to study its ecological and behavioral aspects. This study reports on the karyotype patterns of this species, which represents the first contribution to the cytogenetics of this genus. The karyotype of this species is composed of 2n = 48 chromosomes (6m + 4sm + 36st; NF = 96); the heterochromatic regions are located in centromeric or pericentromeric position and are more pronounced in the nucleolar organizer regions. Two sites Ag-NORs/CMA+/DAPI were identified in the short arms of pairs 2 (metacentric) and 21 (subtelocentric). Unlike the other species of this family which show an evolution modulated by events of centric fusions, H. antenori shows the maintenance of a basal diploid number and the large number of bibrachial elements indicates karyotypic diversification derived by pericentric inversions. Cytogenetic analyzes in this species will provide new taxonomic markers capable of being utilized in conservation issues and systematics. 2015 Reichwald Insights into Sex Chromosome Evolution and Aging from the Genome of a Short-Lived Fish Abstract The killifish Nothobranchius furzeri is the shortest-lived vertebrate that can be bred in the laboratory. Its rapid growth, early sexual maturation, fast aging, and arrested embryonic development (diapause) make it an attractive model organism in biomedical research. Here, we report a draft sequence of its genome that allowed us to uncover an intra-species Y chromosome polymorphism representing—in real time—different stages of sex chromosome formation that display features of early mammalian XY evolution “in action.” Our data suggest that gdf6Y, encoding a TGF-β family growth factor, is the master sex-determining gene in N. furzeri. Moreover, we observed genomic clustering of aging-related genes, identified genes under positive selection, and revealed significant similarities of gene expression profiles between diapause and aging, particularly for genes controlling cell cycle and translation. The annotated genome sequence is provided as an online resource 2016 Kanamori A Genetic Map for the Only Self-Fertilizing Vertebrate Abstract The mangrove killifish Kryptolebias marmoratus, and its close relative Kryptolebias hermaphroditus, are the only vertebrate species known to reproduce by self-fertilization due to functional ovotestis development. To improve our understanding of their genomes, we constructed a genetic map. First, a single F1 fish was made by artificial fertilization between K. marmoratus and K. hermaphroditus strains. F2 progeny were then obtained by self-fertilization of the F1 fish. We used RAD-seq to query genomic DNAs from the two parental strains, the F1 individual and 49 F2 progeny. Results identified 9904 polymorphic RAD-tags (DNA markers) that mapped to 24 linkage groups, corresponding to the haploid chromosome number of these species. The total length of the map was 1248 cM, indicating that about one recombination occurred for each of the 24 homologous chromosome pairs in each meiosis. Markers were not evenly distributed along the chromosomes: in all chromosomes, many markers (> 8% of the total markers for each chromosome) mapped to chromosome tips. Centromeres suppress recombination, and this uneven distribution is probably due to the species’ acrocentric chromosomes. Mapped marker sequences were compared to genomic sequences of medaka and platyfish, the next most closely related species with sequenced genomes that are anchored to genetic maps. Results showed that each mangrove killifish chromosome corresponds to a single chromosome of both platyfish and medaka, suggesting strong conservation of chromosomes over 100 million years of evolution. Our genetic map provides a framework for the K. marmoratus/K. hermaphroditus genome sequence and an important resource for understanding the biology of hermaphroditism. 2016 Krysanov Divergent karyotypes of the annual killifish genus Nothobranchius (Cyprinodontiformes, Nothobranchiidae) Annual killifishes belonging to the genus Nothobranchius Peters, 1868 are mainly distributed in eastern Africa but several species are found in central Africa (Wildekamp 2004). They inhabit temporary pools that dry out during the dry season and have specific adaptations for extreme environments. Annual fishes are characterised by specific life history traits of extremely short lifespan and diapause in embryonic development (Furness 2015, Nagy 2015). Their unique biology makes them a model taxon with which to investigate aging, embryonic development, ecology, and natural selection (Cellerino et al. 2015). Killifishes of the genus Nothobranchius comprise 71 valid species (FishBase 2015). In this genus karyologicaly were described only 23 species (Arai 2011). These species have variable karyotypes with diploid chromosome numbers (2n) ranging from 2n = 16 for Nothobranchius rachovii Ahl, 1926 to 2n = 43 for Nothobranchius thierryi (Ahl, 1924) (Scheel 1990). More than 60% of karyotypes in Nothobranchius are characterised by a modal diploid number of 2n = 36-38. A multiple-sex chromosome system of X1X1X2X2/X1X2Y type has been reported for only one species of Nothobranchius, Nothobranchius guentheri (Pfeffer, 1893) with a female karyotype consisting of 36 chromosomes and the male karyotype consisting of 35 chromosomes (Ewulonu et al. 1985). In this paper, the karyotypes of two species, Nothobranchius brieni Poll, 1938 and Nothobranchius sp. from Kasenga, were studied, bringing the number of species studied to 25. 2016 Waits Genetic Linkage Map and Comparative Genome Analysis for the Atlantic Killifish (Fundulus heteroclitus) Genetic linkage maps are valuable tools in evolutionary biology; however, their availability for wild populations is extremely limited. Fundulus heteroclitus (Atlantic killifish) is a non-migratory estuarine fish that exhibits high allelic and phenotypic diversity partitioned among subpopulations that reside in disparate environmental conditions. An ideal candidate model organism for studying gene-environment interactions, the molecular toolbox for F. heteroclitus is limited. We identified hundreds of novel microsatellites which, when combined with existing microsatellites and single nucleotide polymorphisms (SNPs), were used to construct the first genetic linkage map for this species. By integrating independent linkage maps from three genetic crosses, we developed a consensus map containing 24 linkage groups, consistent with the number of chromosomes reported for this species. These linkage groups span 2300 centimorgans (cM) of recombinant genomic space, intermediate in size relative to the current linkage maps for the teleosts, medaka and zebrafish. Comparisons between fish genomes support a high degree of synteny between the consensus F. heteroclitus linkage map and the medaka and (to a lesser extent) zebrafish physical genome assemblies. 2017-Berois A global community effort to decipher the unique biology of annual killifish Abstract Over the past 50 years, annual killifishes arose as alternative model organisms for studies of vertebrate biology. The annual fish offers exceptional advantages for studies of genetics, genomics, developmental biology, population dynamics, ecology, biogeography, and evolution. They inhabit extremely variable freshwater environments in Africa and South America, have a short lifespan and a set of unique and fascinating developmental characteristics. Embryos survive within the dry substrate during the dry season, whereas the adult population dies. Thus, the survival of the populations is entirely dependent on the buried embryos that hatch the next rainy season. Although Old and New World species share similarities in their life cycle, they also have different adaptive responses associated with climate-related selective pressures. Therefore, contrasting different species from these areas is essential to understand unique adaptations to heterogeneous environment. A network of laboratories (United States, Czech Republic, Italy, Brazil, Chile, and Uruguay) is working and collaborating on many aspects of the biology of annual fishes. Participating researchers share projects and cross-training undergraduate and graduate students. These efforts resulted in two International Symposia (2010 and 2015) that took place in Montevideo and an international book. Herein, we summarize the progress made by this global community of scientists. 2017 Araya-Jaime Chromosomal organization of four classes of repetitive DNA sequences in killifish Orestias ascotanensis Parenti, 1984 (Cyprinodontiformes, Cyprinodontidae) Abstract Orestias Valenciennes, 1839 is a genus of freshwater fish endemic to the South American Altiplano. Cytogenetic studies of these species have focused on conventional karyotyping. The aim of this study was to use classical and molecular cytogenetic methods to identify the constitutive heterochromatin distribution and chromosome organization of four classes of repetitive DNA sequences (histone H3 DNA, U2 snRNA, 18S rDNA and 5S rDNA) in the chromosomes of O. ascotanensis Parenti, 1984, an endemic species restricted to the Salar de Ascotán in the Chilean Altiplano. All individuals analyzed had a diploid number of 48 chromosomes. C-banding identified constitutive heterochromatin mainly in the pericentromeric region of most chromosomes, especially a GC-rich heterochromatic block of the short arm of pair 3. FISH assay with an 18S probe confirmed the location of the NOR in pair 3 and revealed that the minor rDNA cluster occurs interstitially on the long arm of pair 2. Dual FISH identified a single block of U2 snDNA sequences in the pericentromeric regions of a subtelocentric chromosome pair, while histone H3 sites were observed as small signals scattered in throughout the all chromosomes. This work represents the first effort to document the physical organization of the repetitive fraction of the Orestias genome. These data will improve our understanding of the chromosomal evolution of a genus facing serious conservation problems. 2017 Peichel Chromosome Evolution: Molecular Mechanisms and Evolutionary Consequences Long before the advent of “next-generation” or even “first-generation” sequencing, biologists could observe the genome of an organism using relatively simple cytogenetic methods. As a result, we now have information about the number and structure of chromosomes in thousands of organisms. Even at this relatively crude level of observation, it is clear that there is extensive variation in chromosome number and structure among species, and that there can even be dramatic differences in karyotype between closely related species. Cytogenetic studies have also revealed that males and females of many species differ in karyotype with visible sex chromosomes. Understanding both the causes and consequences of such changes in chromosome number and structure has been of long-standing interest to evolutionary biologists. Garcia 1993 Analysis of Karyotypic Evolution in Natural Populations of Cynolebias (Pisces: Cyprinodontiformes, Rivulidae) Using Banding Techniques 2001 Chromosome evolution in the annual killifish genus Cynolebias and mitochondrial phylogenetic analysis 2002 Mitochondrial genes: signals and noise in the phylogenetic reconstruction of the annual killifish genus Cynolebias (Cyprinodontiformes, Rivulidae) 2002a 2006 Multiple simultaneous speciation in killifishes of the Cynolebias adloffi species complex (Cyprinodontiformes, Rivulidae) from phylogeography and chromosome data 2008 High levels of mitochondrial cytochrome b divergence in annual killifishes of the genus Cynolebias (Cyprinodontiformes, Rivulidae) 2014 Burst speciation processes and genomic expansion in the neotropical annual killifish genus Austrolebias (Cyprinodontiformes, Rivulidae) 2015 Next-generation sequencing detects repetitive elements expansion in giant genomes of annual killifish genus Austrolebias (Cyprinodontiformes, Rivulidae) 1972 Scheel Chromosome numbers of fishes Nearly 300 chromosome counts of fishes are listed.