Karyotype differentiation in Chromaphyosemion killifishes (Cyprinodontiformes, Nothobranchiidae): patterns, mechanisms, and evolutionary implications

Chromaphyosemion killifishes are a karyotypically highly diverse group of small, sexually dimorphic fishes living in rainforest rivulets in tropical West and Central Africa. In the present study, we used various chromosome banding and staining techniques to analyse the karyotypes of 13 populations representing seven described species ( Chromaphyosemion loennbergii , Chromaphyosemion punctulatum , Chromaphyosemion splendopleure , Chromaphyosemion volcanum , Chromaphyosemion malumbresi , Chromaphyosemion melanogaster , Chromaphyosemion bitaeniatum ) and two undescribed forms ( Chromaphyosemion cf. lugens , Chromaphyosemion sp. Rio Muni GEMHS00/41). Diploid chromosome numbers (2 n ) and the number of chromosome arms (NF) ranged from 2 n  = 24 in C. malumbresi to 2 n  = 40 in C. bitaeniatum and from NF = 40 in C. volcanum and C. cf. lugens to NF = 54 in one population of C. loennbergii . A tentative XX/XY sex chromosome system was revealed in C. loennbergii , C. melanogaster , C. malumbresi , and Chromaphyosemion sp. Rio Muni GEMHS00/41. Mapping cytogenetic data for all described Chromaphyosemion species onto a recently published mitochondrial DNA phylogeny revealed a complex pattern of chromosomal evolution with several independent reductions of 2 n and independent modifications of NF and nucleolus organizer region phenotypes. Together with the results of preliminary crossing and mate choice experiments, the cytogenetic and molecular phylogenetic data suggest that, contrary to previous hypotheses, chromosomal rearrangements are probably not the most important and certainly not the only factor driving speciation in Chromaphyosemion killifishes.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 94 , 143–153.     

Karyotype differentiation in Chromaphyosemion killifishes (Cyprinodontiformes, Nothobranchiidae). II: cytogenetic and mitochondrial DNA analyses demonstrate karyotype differentiation and its evolutionary direction in C. riggenbachi

African killifishes of the genus Chromaphyosemion show a high degree of phenotypic and karyotypic diversity. The latter is especially pronounced in C. riggenbachi, a morphologically defined species restricted to a small distribution area in Cameroon. This study presents a detailed reconstruction of karyotype differentiation within C. riggenbachi using conventional Giemsa staining and sequential chromosome banding as well as a phylogenetic analysis based on part of the mitochondrial (mt) cytochrome b gene from eleven populations. The cytogenetic analysis revealed differences in chromosome morphology, banding patterns and/or diploid chromosome number (2n) among all populations examined. Diploid number ranged from 2n = 20 to 2n = 36 and varied mainly among populations, while C-banding patterns and NOR phenotypes showed fixed differences among populations as well as some variability within populations. The mtDNA analysis disclosed five clearly differentiated haplotype groups. Mapping the karyotype data onto the mtDNA dendrogram revealed a decrease in 2n from the most basal to the most derived groups, thus demonstrating a reduction of 2n during their evolutionary history. Our results indicate that karyotype differentiation involved Robertsonian fusions as well as non-Robertsonian processes. Causes of the high karyotypic variability may include an elevated chromosomal mutation rate as well as certain features of the ecology and mating system that could facilitate the fixation of chromosomal rearrangements. The pattern of karyotype and haplotype differentiation and the results of previous crossing experiments suggest incipient speciation in C. riggenbachi.     

Female mating preference for own males on species and population level in Chromaphyosemion killifishes (Cyprinodontiformes, Nothobranchiidae)

Chromaphyosemion killifishes are a very promising taxon for the examination of general principles and mechanisms underlying speciation. The polygamous mating system and high degree of sexual dimorphism in this group suggest that sexual selection and female mating preferences play a crucial role in the current radiation process. Because the emergence of mating preferences for individuals of the own population during allopatry is a necessary precondition for speciation by sexual selection, either via diverging mate recognition traits or reinforcement, we examined whether females of different nominal species (Chromaphyosemion bivittatum vs. Chromaphyosemion volcanum) or populations (C. bivittatum “Toko” vs. C. bivittatum “Ilor”) prefer their own males as mating partners. For this purpose, two different methods were used. First, we conducted simultaneous choice tests where females could choose between two different males, and second, we counted the eggs produced by females in consecutive matings with different males. Both approaches produced the same result: females of the populations under examination preferred to mate with their own males. Preference for own males was symmetrical in all tests. Males, on the other hand, do not discriminate between females, not even on the level of nominal species. Thus, our results are in accordance with the hypothesis that female choice is an important factor for speciation in Chromaphyosemion. Additionally, we present some preliminary data on post-zygotic isolation between the populations.     

Comparative morphology,phylogeny, and classification of West African callopanchacine killifishes (Teleostei: Cyprinodontiformes: Nothobranchiidae)

A phylogenetic analysis combining 63 morphological characters and DNA sequences (3296 bp), comprising segments of the mitochondrial genes 16S and ND2, and the nuclear gene 28S, for 19 taxa of the West African killifish tribe Callopanchacini and 11 out‐group taxa, highly supported the monophyly of the tribe, and made it possible to provide the first unambiguous diagnoses for the included genera (Archiaphyosemion, Callopanchax, Nimbapanchax, and Scriptaphyosemion). The monophyly of the Callopanchacini is supported by six morphological synapomorphies: posterior portion of the mandibular channel consisting of a single open groove; basihyal pentagonal, as a result of a nearly rectangular basihyal cartilage and a triangular bony support; dorsal process of the urohyal usually absent, sometimes rudimentary; presence of a wide bony flap adjacent to the proximal portion of the fourth ceratobranchial; a broad bony flap adjacent to the proximal portion of the fifth ceratobranchial; and haemal prezygapophysis of the pre‐ural vertebra 2 ventrally directed. The analysis indicates that the medially continuous rostral neuromast channel, commonly used to diagnose the tribe, is plesiomorphic. This study also indicates that, among African aplocheiloids, the annual life cycle style developed once in Callopanchax, and then again independently in the clade containing Fundulopanchax and Nothobranchius. © 2015 The Linnean Society of London     

High levels of mitochondrial cytochrome b divergence in annual killifishes of the genus Cynolebias (Cyprinodontiformes, Rivulidae)

Phylogenetic relationships based on 324 base pairs of the mitochondrial cytochrome b gene were examined in 14 species of the genus Cynolebias. The monophyly of the genus relative to three outgroup taxa belonging to the family Rivulidae was supported by the sequence data. Bootstrap values corroborated the existence of intrageneric monophyletic units in both parsimony and Neighbour-joining analyses. These include a Cynolebias bellottii-C. wolterstorffl clade, a C. adloJfi-2, C. duraznensis, C. viarius and C adloffi-1 group, and a C. gymnoventris, C. luteoflammulatus pair and the strongly supported assemblage that includes C. prognalhus and C. cheradophilus clade. Phylogenetic relationships remain poorly supported for C. nigripinnis, C. qffinis and C. alexandri, and unresolved among the previous ingroup clades. Cytochrome b sequences reveal an unexpectedly high level of divergence among species of the genus Cynolebias. Consequently, cytochrome b shows good resolution of recent cladogenetic events but limited phylogenetic information at deeper nodes. High levels of sequence divergence span a broad range within Cynolebias. The highest sequence divergence (c. 28%) occurred among C. antenori and the remaining species of the genus. The minimum divergence value (4.5%) is exhibited by sympatric species C. cheradophilus and C. prognathus.     

Intercontinental karyotypic differentiation of Chironomus entis Shobanov, a Holarctic member of the C. plumosus group (Diptera, Chironomidae).

Analysis of banding sequences of polytene chromosomes in Palearctic (Russian) and Nearctic (North American) Chironomus entis shows strong karyotype divergence between populations on the two continents. Four out of seven chromosomal arms in the North American C. entis karyotype are characterized by sequences found only in the Nearctic. In total, 44 banding sequences are now known for this species across the Holarctic, including 22 exclusively Palearctic, 6 Holarctic, and 16 exclusively Nearctic sequences. The degree of cytogenetic differentiation between Palearctic and Nearctic C. entis populations is an order of magnitude greater than differentiation among populations within either continent, but is only one third as great as the cytogenetic distance between the sibling species C. entis and C. plumosus. C. entis is the only sibling species of C. plumosus uncovered during cytological identification of Chironomus species from more than 50 North American lakes, indicating that the plumosus sibling-species group is much smaller in the Nearctic than in the Palearctic, where a dozen sibling species are known. Cytogenetic distance values calculated between Nearctic and Palearctic representatives of both C. entis and its sibling species C. plumosus are similar, but result from different patterns of karyotype divergence. New World C. entis is distinguished from Old World populations by the 16 uniquely Nearctic sequences, four of which occur in the homozygous state. In contrast, North American C. plumosus has fewer uniquely Nearctic sequences, and only one that occurs as a homozygote. However, four chromosomal arms in C. plumosus that are polymorphic in the Palearctic show fixation, or near fixation, of Holarctic sequences in the Nearctic C. plumosus karyotype. Thus, both the fixation of Holarctic sequences, and the occurrence or fixation of distinctly Nearctic sequences, contribute significantly to karyotype divergence. Patterns of karyotype divergence in Palearctic and Nearctic populations of different Holarctic chironomid species are discussed relative to intercontinental cytogenetic differentiation in other dipterans.     

Transspecific variability of mitochondrial NAD-malate dehydrogenase (E.C.: 1.1.1.37) in Primates

Summary The genetic polymorphism of mitochondrial NAD-malate dehydrogenase has been investigated in Primates. 3 enzyme variants could be demonstrated.

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Zusammenfassung Die mitochondriale NAD-Malatdehydrogenase der Primaten zeigt eine genetisch determinierte Variabilität. Es konnten 3 Enzymvarianten nachgewiesen werden.

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Direktor: Prof. Dr. Dr. H. Ritter

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Supported by the Deutsche Forschungsgemeinschaft.     

Galápagos' Opuntia (prickly pear) cacti: extensive morphological diversity, low genetic variability

Due to the pronounced morphological variation and geographical distribution of Galápagos' Opuntia cacti, numerous hypotheses have been advanced regarding their radiation, diversification, and classification. The currently accepted classification is based on morphology and recognizes six species and fourteen varieties, but the plasticity of many of the characteristics renders any morphological taxonomy problematic. Our analysis of previously published morphological data agrees only partially with the current classification. We present the first molecular phylogeny of these plants. Multiple DNA sequences indicate little genetic distinction among the currently identified species, despite restricted gene flow and limited long distance dispersal within the archipelago. No clear relationship exists between morphological and genetic differences. These results suggest that both molecular and morphological data should be used in conservation planning.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 451–461.     

Species delimitation in annual killifishes from the Brazilian Caatinga,the Hypsolebias flavicaudatus complex (Cyprinodontiformes: Rivulidae): implications for taxonomy and conservation

The organisms investigated are members of the Hypsolebias flavicaudatus species complex; a clade of morphologically similar annual killifishes endemic to the semi-arid savannah of north-eastern Brazil in the São Francisco River basin, which is a tropical area under intensive process of habitat loss. A taxonomic revision was conducted combining two different approaches for species delimitation – a tree-based approach using mt-DNA (cytochrome b) and a character-based approach using morphological variation (colour patterns, fin morphology, meristic and morphometric data). Nine species are recognised, of which five are new species: H. flagellatus (Costa), H. flavicaudatus (Costa & Brasil), H. gilbertobrasili sp. nov., H. guanambi (Costa & Amorim), H. janaubensis (Costa), H. nitens sp. nov., H. pterophyllus sp. nov., H. radiseriatus sp. nov. and H. sertanejo sp. nov. On the basis of 17 years of field studies, preliminary conservation status assessments indicate that two species are highly threatened with extinction, at least one is endangered and another is vulnerable, whereas the others are not endangered. This result deeply contrasts with a previous assessment 10 years ago, when the whole H. flavicaudatus complex was considered as a single species of no conservation concern. Discrepancies in the assessment of conservation among closely related species living in similar habitats are a consequence of heterogeneous human occupation and disproportionate recent development throughout the São Francisco River basin.     

Extraordinary and extensive karyotypic variation: a 48-fold range in chromosome number in the gall-inducing scale insect Apiomorpha (Hemiptera: Coccoidea: Eriococcidae).

Chromosome number reflects strong constraints on karyotype evolution, unescaped by the majority of animal taxa. Although there is commonly chromosomal polymorphism among closely related taxa, very large differences in chromosome number are rare. This study reports one of the most extensive chromosomal ranges yet reported for an animal genus. Apiomorpha Rübsaamen (Hemiptera: Coccoidea: Eriococcidae), an endemic Australian gall-inducing scale insect genus, exhibits an extraordinary 48-fold variation in chromosome number with diploid numbers ranging from 4 to about 192. Diploid complements of all other eriococcids examined to date range only from 6 to 28. Closely related species of Apiomorpha usually have very different karyotypes, to the extent that the variation within some species-groups is as great as that across the entire genus. There is extensive chromosomal variation among populations within 17 of the morphologically defined species of Apiomorpha indicating the existence of cryptic species-complexes. The extent and pattern of karyotypic variation suggests rapid chromosomal evolution via fissions and (or) fusions. It is hypothesized that chromosomal rearrangements in Apiomorpha species may be associated with these insects' tracking the radiation of their speciose host genus, Eucalyptus.     

Genetic variation in the African rodent subfamily Otomyinae (Muridae). III. Karyotype and allozymes of the ice rat, Otomys sloggetti robertsi.

Results of chromosome G- and C-banding studies on 12 specimens of the ice rat, Otomys sloggetti robertsi, from three localities and allozyme analyses of seven individuals from two localities are reported. The subspecies karyotype contains 42, mostly acrocentric, chromosomes. No chromosomal variation was detected among animals from these localities. Allozyme analysis tentatively suggested a limited gene flow between the two populations examined.     

Karyotype differentiation in three species of Tripogandra Raf. (Commelinaceae) with different ploidy levels

Most species of the genus Tripogandra (Commelinaceae) are taxonomically poorly circumscribed, in spite of having a relatively stable basic number x = 8. Aiming to estimate the cytological variation among Tripogandra species carrying this base number, several structural karyotypic characters were investigated in the diploid T. glandulosa, the hexaploid T. serrulata, and the octoploid T. diuretica. A careful evaluation of chromosome size and morphology did not reveal clear chromosome homeologies among karyotypes. The mean chromosome size was strongly reduced in the octoploid species, but not in the hexaploid species. They also differed largely in the CMA(+) banding pattern and in the number of 5S and 45S rDNA sites per monoploid chromosome complement. All three species showed proximal DAPI (+) heterochromatin, although in T. serrulata this kind of heterochromatin was only visible after FISH. Further, the meiosis in T. serrulata was highly irregular, suggesting that this species has a hybrid origin. The data indicate that, in spite of the conservation of the base number, these species are karyologically quite different from each other.     

Morphological and karyotypic differentiation in Caranx lugubris (Perciformes: Carangidae) in the St. Peter and St. Paul Archipelago,mid-Atlantic Ridge

Isolated oceanic islands constitute interesting model systems for the study of colonization processes, as several climatic and oceanographic phenomena have played an important role in the history of the marine ichthyofauna. The present study describes the presence of two morphotypes of Caranx lugubris, in the St. Peter and St. Paul Archipelago located in the mid-Atlantic. Morphotypes were compared in regard to their morphological and cytogenetic patterns, using C-banding, Ag-NORs, staining with CMA₃/DAPI fluorochromes and chromosome mapping by dual-color FISH analysis with 5S rDNA and 18S rDNA probes. We found differences in chromosome patterns and marked divergence in body patterns which suggest that different populations of the Atlantic or other provinces can be found in the Archipelago of St. Peter and St. Paul.     

Karyotype of Callithrix mauesi (Callitrichidae,Primates) and its relations with those of C. emiliae and C. jacchus

A study was conducted on the most recently described marmoset species, Callithrix mauesi, and the results obtained were compared to those previously reported for the karyotypes of C. jacchus and C. emiliae. No mechanism of chromosome rearrangement differentiates the karyotypes of C. mauesi (2n = 44) and C. emiliae (2n = 44), which diverge from C. jacchus (2n = 46) by a Robertsonian translocation and a paracentric inversion. C. mauesi, like C. emiliae, presents telomeric constitutive heterochromatin in various chromosomes, forming large heterochromatic blocks in some. This does not occur in C. jacchus, which basically presents centromeric constitutive heterochromatin. The karyotype of C. mauesi differs from that of C. emiliae only by the amount and distribution of this telomeric constitutive heterochromatin. One of the chromosomes presenting a heterochromatic block in C. mauesi is chromosome X, a fact not previously reported in the Order Primates. The present chromosome data show that C. mauesi is closer to C. emiliae than to C. jacchus, in agreement with its inclusion in the C. argentata group. In the present paper, we describe for the first time, at the chromosome level, chimerism between fraternal twins of the same sex (XY/XY), with the heterochromatic block of pair 2 being the marker. © 1994 Wiley-Liss, Inc.     

An early stage of ZW/ZZ sex chromosome differentiation in Poecilia sphenops var. melanistica (Poeciliidae,Cyprinodontiformes)

The mitotic and meiotic chromosomes of the American cyprinodont fish Poecilia sphenops var. melanistica were analysed. All 46 chromosomes are telocentric. By specific staining of the constitutive heterochromatin with C-banding and various AT-specific fluorochromes, the homomorphic chromosome pair 1 could be identified as sex chromosomes of the ZW/ZZ type. All female animals exhibit a W chromosome with a large region of telomeric heterochromatin that is not present in the Z chromosome. These sex chromosomes cannot be distinguished by conventional staining; they represent the first demonstration of sex chromosomes in fishes in an early stage of morphological differentiation. The W heterochromatin and the telomeric heterochromatin in the two autosomes 18 show a very bright fluorescence when stained with AT-specific fluorochromes. This allows the direct identification of the chromosomal sex by examining the interphase nuclei: females exhibit three, males only two brightly fluorescent heterochromatic chromocenters in their nuclei. The significance of these ZW/ ZZ sex chromosomes and their specific DNA sequences, the dose compensation of the Z-linked genes, and the experimental possibilities using sex-reversed ZW males are discussed.