Cytogenetics of the razor clam Solen marginatus (Mollusca: Bivalvia: Solenidae)

The razor clam Solen marginatus has a diploid chromosome number of 38. The karyotype consists of one metacentric/submetacentric, three submetacentric/metacentric, five submetacentric, one submetacentric/subtelocentric, one subtelocentric/submetacentric, six subtelocentric and two telocentric chromosome pairs. Staining with chromomycin A3 revealed bright positive bands subcentromerically in the long arms of one medium-sized subtelocentric pair, while DAPI staining showed uniform fluorescence in all chromosomes of the complement. Fluorescence in situ hybridization using an 18S-5.8S-28S rDNA probe locates these loci at the subcentromeric region of one subtelocentric pair and at the subtelomeric region of another subtelocentric pair.     

Integrated gene mapping and synteny studies give insights into the evolution of a sex proto-chromosome in <Emphasis Type="Italic">Solea senegalensis</Emphasis>

The evolution of genes related to sex and reproduction in fish shows high plasticity and, to date, the sex determination system has only been identified in a few species. Solea senegalensis has 42 chromosomes and an XX/XY chromosome system for sex determination, while related species show the ZZ/ZW system. Next-generation sequencing (NGS), multi-color fluorescence in situ hybridization (mFISH) techniques, and bioinformatics analysis have been carried out, with the objective of revealing new information about sex determination and reproduction in S. senegalensis. To that end, several bacterial artificial chromosome (BAC) clones that contain candidate genes involved in such processes (dmrt1, dmrt2, dmrt3, dmrt4, sox3, sox6, sox8, sox9, lh, cyp19a1a, amh, vasa, aqp3, and nanos3) were analyzed and compared with the same region in other related species. Synteny studies showed that the co-localization of dmrt1-dmrt2-drmt3 in the largest metacentric chromosome of S. senegalensis is coincident with that found in the Z chromosome of Cynoglossus semilaevis, which would potentially make this a sex proto-chromosome. Phylogenetic studies show the close proximity of S. senegalensis to Oryzias latipes, a species with an XX/XY system and a sex master gene. Comparative mapping provides evidence of the preferential association of these candidate genes in particular chromosome pairs. By using the NGS and mFISH techniques, it has been possible to obtain an integrated genetic map, which shows that 15 out of 21 chromosome pairs of S. senegalensis have at least one BAC clone. This result is important for distinguishing those chromosome pairs of S. senegalensis that are similar in shape and size. The mFISH analysis shows the following co-localizations in the same chromosomes: dmrt1-dmrt2-dmrt3, dmrt4-sox9-thrb, aqp3-sox8, cyp19a1a-fshb, igsf9b-sox3, and lysg-sox6.     

Un nouveau système chromosomique polymorphe chez desLeggada africaines du groupeTenellus (Rodentia-Muridae)

The pigmy-mice belonging to the grouptenellus seem fairly well characterized through the structure of the teeth (M₁). However, the chromosome complement of specimens from Niamey (Niger) is strikingly different from this precedently described bytenellus from Accra (Ghana). In this last station, we had to deal with the “primitive”, complement (2N=36,N.F.=36, sex-chromosomesPR). The sample of Niamey has the following constitution: three males have 33, 33 and 32 chromosomes, the single female has 34. This robertsonian polymorphism is due to the presence either of four acrocentric autosomes, or of one submetacentric and two acrocentrics, or of two submetacentrics. This polymorphism is fundamentally alike to that demonstrated precedently in the groupminutoides. The sex-chromosomes are of primitive type (PR). Two autosomes of medium size are satellited and their metacentricity seems to result from a pericentric inversion increasing theN.F. of two, 38 instead 36.     

Karyotype of the sculpin <Emphasis Type="Italic">Myoxocephalus jaok</Emphasis> (Pisces: Cottidae) from Peter the Great Bay,Sea of Japan

The karyotype of the sculpin Myoxocephalus jaok from the Ussuriisky, Amursky, and Vostok Bays was studied. It was found that 2n = 24, among the chromosomes there were 16 metacentric, 4 submetacentric, and 4 acrocentric, NF = 44. There was no variation in the number of chromosomes, and no differences between male and female karyotypes were revealed. A comparison of Myoxocephalus jaok, M. brandti and M. stelleri karyotypes was performed. The pronounced distinction of Myoxocephalus jaok karyotype resulting from chromosomal rearrangements was shown.     

Comparative cytogenetic analysis of four species of <Emphasis Type="Italic">Dendropsophus</Emphasis> (Hylinae) from the Brazilian Atlantic forest

We conducted a cytogenetic study of four hyline frog species (Dendropsophus elegans, D. microps, D. minutus and D. werneri) from southern Brazil. All species had 2n = 30 chromosomes, with interspecific and intraspecific variation in the numbers of metacentric, submetacentric, subtelocentric and telocentric chromosomes. C-banding and fluorochrome staining revealed conservative GC-rich heterochromatin localized in the pericentromeric regions of all species. The location of the nucleolus organizer regions, as confirmed by fluorescent in situ hybridization, differed between species. Telomeric probes detected sites that were restricted to the terminal regions of all chromosomes and no interstitial or centromeric signals were observed. Our study corroborates the generic synapomorphy of 2n = 30 chromosomes for Dendropsophus and adds data that may become useful for future taxonomic revisions and a broader understanding of chromosomal evolution among hylids.     

The Karyotype of the flathead sculpin Megalocottus platycephalus taeniopterus (Kner, 1868) (Pisces: Cottidae) from Vostok Bay,Sea of Japan

For the first time, we studied the karyotype of the flathead sculpin Megalocottus platycephalus taeniopterus (Kner, 1868) from Vostok Bay of the Sea of Japan. The karyotype is stable: 2n = 42 (2 metacentric, 2 submeta-subtelocentric, 30 subtelocentric, and 8 acrocentric chromosomes), NF = 44 + 2. The nucleolar organizers (NOs) were identified using Ag banding in two pairs of chromosomes: in the telomeric parts of the short arm of the medium-size subtelocentric chromosome and the long arm of the large acrocentric chromosome. Variations in the number of nucleolar organizer chromosomes and in the number of NO staining blocks were found. Comparison of the karyotypes of M. p. taeniopterus and previously studied M. p. platycephalus (Pallas, 1814) from the northern Sea of Okhotsk revealed their similarity in the number and morphology of chromosomes and the number of chromosome arms and difference between the subspecies in the number and location of NO, which allows their discrimination.     

The karyotype of the great sculpin, <Emphasis Type="Italic">Myoxocephalus polyacanthocephalus</Emphasis> (Pallas, 1814) (Pisces: Cottidae), from the Russian part of the species range

The karyotype of the great sculpin, Myoxocephalus polyacanthocephalus (Pallas, 1814) (Pisces: Cottidae) from the Sea of Okhotsk and the Sea of Japan has been studied for the first time. The karyotype is stable; it consists of 40 chromosomes (4 metacentric, 2 submeta-subtelocentric, 20 subtelocentric, and 14 acrocentric chromosomes); the number of chromosomal arms is 44 + 2. Nucleolar organizer regions (NOR) are found in the telomeric region of the arm in one homologue of a pair of small metacentric chromosomes, using the Ag-NOR banding technique. A comparative analysis of the karyotype of M. polyacanthocephalus and the karyotypes of other Myoxocephalus species (M. stelleri, M. brandtii, M. jaok, M. ochotensis, and M. scorpius) has been carried out based on the main karyotype characters, as well as on the number and localization of NORs. The identified differences make it possible to differentiate the studied species, whereas the general traits indicate their taxonomic proximity.     

Somatic chromosomes of squirrel monkey (Saimiri sciureus)

The chromosome complement of 2 male and 15 female adult squirrel monkeys (Saimiri sciureus) have been studied in peripheral blood and kidney cultures and bone marrow preparations. The diploid chromosome number is 44. Six of the chromosome pairs are metacentric (isobrachial), 9 pairs submetacentric (heterobrachial) and the remaining 6 pairs are acrocentric (cephalobrachial). The X chromosome is the longest submetacentric (heterobrachial), ranking 5 in order of decreasing size. The Y chromosome is the smallest acrocentric (cephalobrachial) of the complement.A female and male karyotype is presented.Supported, in part, by Grant HDO 1952-03 from the United States Public Health Service.     

Chromosome identification for the carnivorous plant <Emphasis Type="Italic">Genlisea margaretae</Emphasis>

Genlisea margaretae, subgenus Genlisea, section Recurvatae (184 Mbp/1C), belongs to a plant genus with a 25-fold genome size difference and an extreme genome plasticity. Its 19 chromosome pairs could be distinguished individually by an approach combining optimized probe pooling and consecutive rounds of multicolor fluorescence in situ hybridization (mcFISH) with bacterial artificial chromosomes (BACs) selected for repeat-free inserts. Fifty-one BACs were assigned to 18 chromosome pairs. They provide a tool for future assignment of genomic sequence contigs to distinct chromosomes as well as for identification of homeologous chromosome regions in other species of the carnivorous Lentibulariaceae family, and potentially of chromosome rearrangements, in cases where more than one BAC per chromosome pair was identified.     

马尾松染色体荧光带型的研究

对马尾松有丝分裂中期染色体荧光带纹的分析结果表明,其色霉素Ａ的染色体的荧光带赤;１对为着丝粒区和臂间我均有带纹的中间着丝染色体。６对为臂间区有带纺的中间着丝粒染色体;２对为着丝粒区有带纹的中间着丝粒染色体;３对无带纹的中间或近中着丝粒染色体;１对为着丝粒区有带纹的近中着丝粒染色体。     

Dual-color FISH karyotype analyses using rDNAs in three Cucurbitaceae species

Dual-color fluorescence in situ hybridization (FISH) analysis of three Cucurbitaceae species from different genera was conducted using 5S and 45S rDNA probes. In Benincasa hispida (Thunb.) Cogn. (2n=24), the 45S rDNA probe hybridized on two chromosomes, one in the short arm of a medium-sized metacentric chromosome and another at the satellite of a chromosome. The 5S rDNA hybridized at a site proximal to the centromere of the same short arm of the 45S rRNA gene locus that occupied almost the entire short arm. For Citrullus lanatus (Thunb.) Matsum & Nakai (2n=22), the 45S rDNA probe hybridized at sites in the short arms of two chromosomes and the 5S rDNA probe was co-localized with the 45S rRNA locus at the region proximal to the centromere in one chromosome. The 45S rRNA loci occupied almost all of the short arms in both chromosomes. In Cucurbita moschata Duch. (2n=40), the 45S rDNA probe hybridized in five chromosomes in which the 45S rRNA genes occupied almost two-thirds of the chromosomes in two large chromosomes and the entire short arm of a medium-sized chromosome. Two other loci were present in two medium-sized chromosomes, one in the proximal region in the short arm of a chromosome and another at the tip of the long arm of a chromosome. Chromosomes of B. hispida were relatively larger than those of the other two species. The karyotype of B. hispida is composed of two metacentrics and 10 submetacentrics, while that of C. lanatus is composed of seven metacentrics and four submetacentrics and that of C. moschata is composed of 18 metacentrics and two submetacentrics. Comparative chromosome evolution among the three Cucurbitaceae species was attempted using the karyotypes and the chromosomal distribution patterns of the 5S and 45S rDNAs. The results presented herein will be useful in elucidating the phylogenetic relationships among Cucurbitaceae species, and will provide basic data for their breeding programs.     

Cytogenetic analyses of eight species in the genus <Emphasis Type="Italic">Leptodactylus</Emphasis> Fitzinger, 1843 (Amphibia,Anura, Leptodactylidae), including a new diploid number and a karyotype with multiple translocations

Background

The karyotypes of Leptodactylus species usually consist of 22 bi-armed chromosomes, but morphological variations in some chromosomes and even differences in the 2n have been reported. To better understand the mechanisms responsible for these differences, eight species were analysed using classical and molecular cytogenetic techniques, including replication banding with BrdU incorporation.

Results

Distinct chromosome numbers were found: 2n = 22 in Leptodactylus chaquensis, L. labyrinthicus, L. pentadactylus, L. petersii, L. podicipinus, and L. rhodomystax; 2n = 20 in Leptodactylus sp. (aff. podicipinus); and 2n = 24 in L. marmoratus. Among the species with 2n = 22, only three had the same basic karyotype. Leptodactylus pentadactylus presented multiple translocations, L. petersii displayed chromosome morphological discrepancy, and L. podicipinus had four pairs of telocentric chromosomes. Replication banding was crucial for characterising this variability and for explaining the reduced 2n in Leptodactylus sp. (aff. podicipinus). Leptodactylus marmoratus had few chromosomes with a similar banding patterns to the 2n = 22 karyotypes. The majority of the species presented a single NOR-bearing pair, which was confirmed using Ag-impregnation and FISH with an rDNA probe. In general, the NOR-bearing chromosomes corresponded to chromosome 8, but NORs were found on chromosome 3 or 4 in some species. Leptodactylus marmoratus had NORs on chromosome pairs 6 and 8. The data from C-banding, fluorochrome staining, and FISH using the telomeric probe helped in characterising the repetitive sequences. Even though hybridisation did occur on the chromosome ends, telomere-like repetitive sequences outside of the telomere region were identified. Metaphase I cells from L. pentadactylus confirmed its complex karyotype constitution because 12 chromosomes appeared as ring-shaped chain in addition to five bivalents.

Conclusions

Species of Leptodactylus exhibited both major and minor karyotypic differences which were identified by classical and molecular cytogenetic techniques. Replication banding, which is a unique procedure that has been used to obtain longitudinal multiple band patterns in amphibian chromosomes, allowed us to outline the general mechanisms responsible for these karyotype differences. The findings also suggested that L. marmoratus, which was formerly included in the genus Adenomera, may have undergone great chromosomal repatterning.

     

Chromosome banding study of the golden loach,Sabanejewia aurata balcanica from Slovakia (Cobitidae)

The karyotype of a subspecies of the golden loach,Sabanejewia aurata balcanica from eastern Slovakia was studied by conventional Giemsa staining, Ag-NOR staining, and C-banding. The diploid chromosome number was 2n = 50. The karyotype comprised 2 pairs of metacentric, 6 pairs of submetacentric and 17 pairs of subtelocentric to acrocentric chromosomes. Both metacentric pairs and 2 large subtelocentric pairs had massive pericentromeric blocks, while all other elements had only weak blocks of heterochromatin. The NORs were localized on the short arms of one middle-sized subtelocentric pair. The karyotype ofS. a. balcanica differs from that ofS. aurata kubanica, suggesting chromosomal polymorphism of this widely distributed, polytypic cobitid species. The polymorphic karyotypes of the golden loach may thus demonstrate transient stages, linking primitive and advanced cobitid karyotypes.     

Chromosomal polymorphism in <Emphasis Type="Italic">Steindachneridion</Emphasis><Emphasis Type="Italic">melanodermatum</Emphasis> Garavello, 2005 (Siluriformes,Pimelodidae): a reappraisal the existence of sex chromosome system in the species

Fifty-five specimens of Steindachneridion melanodermatum were analyzed using molecular and conventional cytogenetic tools. Two polymorphisms were found: one involving the length of nucleolar organizer regions and another involving two submetacentric chromosomes previously identified as sex chromosomes. The polymorphism was confirmed by homogeneity between male and female karyotypes. Nucleotide sequencing and physical chromosome mapping were also used to identify and characterize one class of repetitive DNA, named SmAluI-Rex3. Based on the results and literature the present study offers an update of the occurrence of sex chromosome system in this species.     

Reconstruction of chromosome rearrangements between the two most ancestral duckweed species <Emphasis Type="Italic">Spirodela polyrhiza</Emphasis> and <Emphasis Type="Italic">S. intermedia</Emphasis>

The monophyletic duckweeds comprising five genera within the monocot order Alismatales are neotenic, free-floating, aquatic organisms with fast vegetative propagation. Some species are considered for efficient biomass production, for life stock feeding, and for (simultaneous) wastewater phytoremediation. The ancestral genus Spirodela consists of only two species, Spirodela polyrhiza and Spirodela intermedia, both with a similar small genome (~160 Mbp/1C). Reference genome drafts and a physical map of 96 BACs on the 20 chromosome pairs of S. polyrhiza strain 7498 are available and provide useful tools for further evolutionary studies within and between duckweed genera. Here we applied sequential comparative multicolor fluorescence in situ hybridization (mcFISH) to address homeologous chromosomes in S. intermedia (2n = 36), to detect chromosome rearrangements between both species and to elucidate the mechanisms which may have led to the chromosome number alteration after their evolutionary separation. Ten chromosome pairs proved to be conserved between S. polyrhiza and S. intermedia, the remaining ones experienced, depending on the assumed direction of evolution, translocations, inversion, and fissions, respectively. These results represent a first step to unravel karyotype evolution among duckweeds and are anchor points for future genome assembly of S. intermedia.     

Comparative cytomolecular analyses reveal karyotype variability related to biogeographic and species richness patterns in Bombacoideae (Malvaceae)

Bombacoideae is one out of nine subfamilies of Malvaceae and encompasses 160 tree species. The subfamily is karyotypically characterized by small and numerous chromosomes and is traditionally known by a remarkable inter- and intraspecific chromosome number variation. We conducted a comparative cytogenetic analysis to investigate karyotype diversity and chromosome evolution within Bombacoideae. To achieve this, we performed new chromosome counts, CMA/DAPI double staining, genome size estimations, and localization of 5S and 45S rDNA by fluorescence in situ hybridization for 21 species distributed across the Bombacoideae phylogeny. We performed ancestral states reconstruction analyses to elucidate chromosome evolution and provide insights into the systematics and evolution of Bombacoideae in comparison with other Malvaceae species. Newly generated data on chromosome number on Bombacoideae revealed diploids (Ochroma (2n = 84), Cavanillesia, Pochota, Pseudobombax (2n = 88), and Pachira (2n = 92)) and polyploids (Adansonia digitata (2n = 160) and Eriotheca species (2n = ca. 194 and 2n = 276)). For most species, in situ hybridization revealed karyotype, with two pairs of 45S rDNA sites co-located with CMA⁺ bands, and 5S rDNA sites in only one chromosome pair. Taken together, our results provide support to the hypothesis of karyotypic stability in Bombacoideae. Only the Pachira s.l. clade displayed some variability in ploidy level, number of CMA⁺ bands and 45S rDNA sites, and genome size compared to other Bombacoideae clades. The Striated bark clade was characterized by comparatively small genomes and low cytomolecular variability. Karyotypic data were related to biogeographic and species richness patterns of Bombacoideae.     

Screening and identification of a microsatellite marker associated with sex in Wami tilapia, <Emphasis Type="Italic">Oreochromis urolepis hornorum</Emphasis>

In this study, primer pairs of 15 microsatellite markers associated with sex determination of tilapia were selected and amplified in Wami tilapia, Oreochromis urolepis hornorum. While one marker, UNH168, on linkage group 3 (LG3) was associated (P < 0.001) with the phenotypic sex in the experimental population, nine genotypes were detected in both sexes. Only 99-bp allele was detected in the female samples, while 141, 149 and 157-bp alleles were present in both male and female samples. UNH168 was localized by fluorescence in situ hybridization (FISH) on the long arm of the largest tilapia chromosome pair (chromosome 1, equivalent to LG3). This sex-linked microsatellite marker could potentially be used for marker-assisted selection in tilapia breeding programmes to produce monosex male tilapia.     

The chromosome morphology ofArabidopsis thaliana (L.)Heynh. and some remarks on the problem ofHylandra suecica (Fr.) Löve

The chromosome number ofA. thaliana from three localities in Central Bohemia was found to be 2n=10. All the chromosomes (length 1,5–2,6μm) belong to the atelocentric type, four pairs (m) having the centromere in the median and one pair (sm) in the submedian region. In connection with the discussion on the origin ofH. suecica the author presents the following preliminary results: a) the failure to cross the tetraploidCardaminopsis arenosa (L.)Hayek withA. thaliana; b) the successful crossing of the diploidC. petraea (L.)Hiit. withA. thaliana; c) the discovery of a diploid population ofC. arenosa (2n=16) in the Tatra Mts. (Czechoslovakia).     

Discovery of the youngest sex chromosomes reveals first case of convergent co-option of ancestral autosomes in turtles

Most turtle species possess temperature-dependent sex determination (TSD), but genotypic sex determination (GSD) has evolved multiple times independently from the TSD ancestral condition. GSD in animals typically involves sex chromosomes, yet the sex chromosome system of only 9 out of 18 known GSD turtles has been characterized. Here, we combine comparative genome hybridization (CGH) and BAC clone fluorescent in situ hybridization (BAC FISH) to identify a macro-chromosome XX/XY system in the GSD wood turtle Glyptemys insculpta (GIN), the youngest known sex chromosomes in chelonians (8–20 My old). Comparative analyses show that GIN-X/Y is homologous to chromosome 4 of Chrysemys picta (CPI) painted turtles, chromosome 5 of Gallus gallus chicken, and thus to the X/Y sex chromosomes of Siebenrockiella crassicollis black marsh turtles. We tentatively assign the gene content of the mapped BACs from CPI chromosome 4 (CPI-4) to GIN-X/Y. Chromosomal rearrangements were detected in G. insculpta sex chromosome pair that co-localize with the male-specific region of GIN-Y and encompass a gene involved in sexual development (Wt1—a putative master gene in TSD turtles). Such inversions may have mediated the divergence of G. insculpta sex chromosome pair and facilitated GSD evolution in this turtle. Our results illuminate the structure, origin, and evolution of sex chromosomes in G. insculpta and reveal the first case of convergent co-option of an autosomal pair as sex chromosomes within chelonians.     

湘华鲮(♂)×鲮鱼(♀)杂交一代与其双亲染色体组型的比较研究

三种鱼的二倍体数目完全相同,均为2n=50。三种鱼的核型差异较大,尤其是双亲间的组型在形态上有明显的差异。然而杂交一代的m、sm二组染色体的组型却十分接近母体,这二组的染色体数占了全部染色体总数的88%。杂交一代染色体组型中的st和t二组染色体虽与双亲有所不同,但与双亲中这二组染色体组型的差异来比要小一些。三种鱼的中期分裂相中,未曾发现带有随体的染色体,也未发现与性别有关的异型染色体的存在。