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Symbiotic relationships between eukaryotes and prokaryotes played pivotal roles in the evolution of life and drove the emergence of specialized symbiotic structures in animals, plants and fungi. The host-evolved symbiotic structures of microbial eukaryotes - the vast majority of such hosts in nature - remain largely unstudied. Here we describe highly structured symbiosomes within three free-living anaerobic protists (Anaeramoeba spp.). We dissect this symbiosis using complete genome sequencing and transcriptomics of host and symbiont cells coupled with fluorescence in situ hybridization, and 3D reconstruction using focused-ion-beam scanning electron microscopy. The emergence of the symbiosome is underpinned by expansion of gene families encoding regulators of membrane trafficking and phagosomal maturation and extensive bacteria-to-eukaryote lateral transfer. The symbionts reside deep within a symbiosomal membrane network that enables metabolic syntrophy by precisely positioning sulfate-reducing bacteria alongside host hydrogenosomes. Importantly, the symbionts maintain connections to the Anaeramoeba plasma membrane, blurring traditional boundaries between ecto- and endosymbiosis.
The cytogenetics of Acanthocephala is a neglected area in the study of this group of endoparasites. Chromosome number and/or karyotypes are known for only 12 of the 1,270 described species, and molecular cytogenetic data are limited to rDNA mapping in two species. The standard karyological technique and mapping of 18S rRNA and H3 histone genes on the chromosomes of Acanthocephalus anguillae individuals from three populations, one of which originated from the unfavorable environmental conditions of the Zemplínska Šírava reservoir in eastern Slovakia, were applied for the first time. All specimens had 2n = 7/8 (male/female); n = 1m + 1m-sm + 1a + 1a (X). Fluorescence in situ hybridization (FISH) revealed three loci of 18S rDNA on two autosomes and dispersion of H3 histone genes on all autosomes and the X chromosome. In addition to the standard A chromosome set, 34% of specimens from Zemplínska Šírava possessed a small acrocentric B chromosome, which was always found to be univalent, with no pairing observed between the B chromosome and the A complement. The B chromosome had a small amount of heterochromatin in the centromeric and telomeric regions of the chromosomal arms and showed two clusters of H3 genes. It is well known that an environment permanently polluted with chemicals leads to an increased incidence of chromosomal rearrangements. As a possible scenario for the B chromosome origin, we propose chromosomal breaks due to the mutagenic effect of pollutants in the aquatic environment. The results are discussed in comparison with previous chromosome data from Echinorhynchida species.
TITLE: Analyse chromosomique et présence de chromosomes B chez le parasite de poisson Acanthocephalus anguillae (Palaeacanthocephala, Echinorhynchida). ABSTRACT: La cytogénétique des Acanthocephala est un domaine négligé dans l’étude de ce groupe d’endoparasites. Le nombre de chromosomes et/ou les caryotypes ne sont connus que pour 12 des 1270 espèces décrites, et les données cytogénétiques moléculaires se limitent à la cartographie de l’ADNr chez deux espèces. La technique caryologique standard et la cartographie des gènes de l’ARNr 18S et de l’histone H3 ont été appliquées pour la première fois sur les chromosomes d’individus d’Acanthocephalus anguillae provenant de trois populations, dont l’une dans les conditions environnementales défavorables du réservoir de Zemplínska Šírava dans l’est de la Slovaquie. Tous les spécimens avaient 2n = 7/8 (mâle/femelle); n = 1m + 1m-sm + 1a + 1a (X). La technique FISH a révélé trois locus d’ADNr 18S sur deux autosomes et une dispersion des gènes de l’histone H3 sur tous les autosomes et sur le chromosome X. En plus de l’ensemble standard de chromosomes A, 34 % des spécimens de Zemplínska Šírava possédaient un petit chromosome B acrocentrique, qui s’est toujours révélé univalent, sans aucun appariement observé entre le chromosome B et le complément A. Le chromosome B avait une petite quantité d’hétérochromatine dans les régions centromériques et télomériques des bras chromosomiques et présentait deux groupes de gènes H3. Il est bien connu qu’un environnement pollué en permanence par des produits chimiques entraîne une incidence accrue de réarrangements chromosomiques. Comme scénario possible pour l’origine du chromosome B, nous proposons des cassures chromosomiques dues à l’effet mutagène des polluants du milieu aquatique. Les résultats sont discutés en comparaison avec les données chromosomiques précédentes des espèces d’Echinorhynchida.
- Klíčová slova
- 18S rDNA, B chromosomes, Fluorescence in situ hybridization, H3 histone, Karyotype, PCB pollution,
- MeSH
- Acanthocephala * genetika MeSH
- histony genetika MeSH
- hybridizace in situ fluorescenční metody MeSH
- karyotyp MeSH
- lidé MeSH
- mapování chromozomů metody MeSH
- paraziti * genetika MeSH
- ribozomální DNA genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- histony MeSH
- ribozomální DNA MeSH
A protocol for primed in situ DNA labeling (PRINS) was optimized for pea (Pisum sativum L.) and field bean (Vicia faba L.) chromosomes attached to coverslips. Cloned DNA or synthetic oligonucleotides were used as probes for repetitive DNA sequences (rDNA, Fok-element) and different reaction conditions were tested to achieve the highest specific signal-to-background ratio. A procedure based on direct labeling by fluorescein-dUTP was compared with an indirect one using digoxigenin detected by fluorescently labeled antibody. Under optimal conditions, strong and specific signals were obtained exclusively on chromosome regions known to contain respective DNA sequences. Compared to the direct labeling, significantly stronger signals were obtained when the indirect procedure was used. Both types of labeling were successfully applied to chromosomes in suspension and were shown to produce signals comparable to that obtained with chromosomes attached to coverslips. It is expected that primed in situ DNA labeling en suspension (PRINSES) will provide a basis for flow-cytometric discrimination and sorting of otherwise indistinguishable chromosomes according to their specific fluorescent labeling.
- MeSH
- deoxyuracilnukleotidy MeSH
- DNA primery * MeSH
- DNA rostlinná analýza MeSH
- DNA sondy MeSH
- Fabaceae genetika MeSH
- fluorescein MeSH
- fluoresceiny MeSH
- fluorescenční mikroskopie MeSH
- hybridizace in situ MeSH
- léčivé rostliny * MeSH
- molekulární sekvence - údaje MeSH
- organizátor jadérka MeSH
- polymerázová řetězová reakce MeSH
- repetitivní sekvence nukleových kyselin MeSH
- sekvence nukleotidů MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- technické zprávy MeSH
- Názvy látek
- deoxyuracilnukleotidy MeSH
- deoxyuridine triphosphate MeSH Prohlížeč
- DNA primery * MeSH
- DNA rostlinná MeSH
- DNA sondy MeSH
- fluorescein MeSH
- fluoresceiny MeSH
The Asian box turtle genus Cuora currently comprises 13 species with a wide distribution in Southeast Asia, including China and the islands of Indonesia and Philippines. The populations of these species are rapidly declining due to human pressure, including pollution, habitat loss, and harvesting for food consumption. Notably, the IUCN Red List identifies almost all species of the genus Cuora as Endangered (EN) or Critically Endangered (CR). In this study, we explore the karyotypes of 10 Cuora species with conventional (Giemsa staining, C-banding, karyogram reconstruction) and molecular cytogenetic methods (in situ hybridization with probes for rDNA loci and telomeric repeats). Our study reveals a diploid chromosome number of 2n = 52 chromosomes in all studied species, with karyotypes of similar chromosomal morphology. In all examined species, rDNA loci are detected at a single medium-sized chromosome pair and the telomeric repeats are restricted to the expected terminal position across all chromosomes. In contrast to a previous report, sex chromosomes are neither detected in Cuoragalbinifrons nor in any other species. Therefore, we assume that these turtles have either environmental sex determination or genotypic sex determination with poorly differentiated sex chromosomes. The conservation of genome organization could explain the numerous observed cases of interspecific hybridization both within the genus Cuora and across geoemydid turtles.
- Klíčová slova
- C-banding, Cuora, FISH, Geoemydidae, evolution, heterochromatin, karyotype, microsatellites, rDNA, telomeres,
- MeSH
- cytogenetické vyšetření * MeSH
- fylogeneze MeSH
- hybridizace in situ fluorescenční MeSH
- karyotyp MeSH
- mikrosatelitní repetice MeSH
- molekulární evoluce MeSH
- pruhování chromozomů MeSH
- ribozomální DNA genetika MeSH
- taxonomické DNA čárové kódování MeSH
- telomery MeSH
- želvy klasifikace genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- ribozomální DNA MeSH
The abundance and chromosomal organization of two repetitive sequences named 12-13P and 18-24J were analyzed in 24 diploid and nine polyploid species of Chenopodium s.l., with special attention to Chenopodium s.s. Both sequences were predominantly present in species of Chenopodium s.s.; however, differences in the amplification levels were observed among the species. The 12-13P repeat was highly amplified in all of the analyzed Eurasian species, whereas the American diploids showed a marked variation in the amplification levels. The 12-13P repeat contains a tandemly arranged 40 bp minisatellite element forming a large proportion of the genome of Chenopodium (up to 3.5%). FISH revealed its localization to the pericentromeric regions of the chromosomes. The chromosomal distribution of 12-13P delivered additional chromosomal marker for B-genome diploids. The 18-24J repeat showed a dispersed organization in all of the chromosomes of the analyzed diploid species and the Eurasian tetraploids. In the American allotetraploids (C. quinoa, C. berlandieri) and Eurasian allohexaploids (e.g., C. album) very intense hybridization signals of 18-24J were observed only on 18 chromosomes that belong to the B subgenome of these polyploids. Combined cytogenetic and molecular analyses suggests that reorganization of these two repeats accompanied the diversification and speciation of diploid (especially A genome) and polyploid species of Chenopodium s.s.
- Klíčová slova
- ADN répété, Chenopodium, fluorescence in situ hybridization, genomic organization, hybridation in situ en fluorescence, organisation génomique, repetitive DNA,
- MeSH
- Amaranthaceae genetika MeSH
- amplifikace genu * MeSH
- genom rostlinný * MeSH
- minisatelitní repetice MeSH
- molekulární evoluce * MeSH
- ploidie MeSH
- Publikační typ
- časopisecké články MeSH
We present a series of 14 cases of cutaneous hidradenocarcinomas. The patients included 6 women and 8 men ranging in age at diagnosis from 34 to 93 years. All but 1 patient presented with a solitary nodule. There was no predilection site. One patient presented with multiple lesions representing metastatic nodules. Of 12 patients with available follow-up, 2 died of disease, whereas the remaining 10 patients were alive but 3 of them experienced a local recurrence in the course of the disease. Grossly, the tumors ranged in size from 1.2 to 6 cm. Microscopically, of the 14 primary tumors, 9 showed low-grade cytomorphology, whereas the remaining 5 neoplasms were high-grade lesions. The residuum of a hidradenoma was present in 5 of the 14 primaries. The mitotic rate was highly variable, ranging from 2 to 64 mitoses per 10 high-power field. The cellular composition of the tumors varied slightly, with clear cells, epidermoid cells, and transitional forms being present in each case. In 1 case, there was metaplastic transformation into sarcomatoid carcinoma. Glandular differentiation varied from case to case and appeared most commonly as simple round glands or as cells with intracytoplasmic lumens. Necrosis en masse was detected in 8 specimens. One specimen represented a reexcision and was unusual as it showed a well-demarcated intradermal proliferation of relatively bland clear cells accompanied by an overlying intraepidermal growth of clear cells resembling hidradenoacanthoma simplex. Despite the bland appearance, the tumor metastasized to a lymph node. Immunohistochemically, 5 of the 8 specimens studied for Her2/neu expression were negative, whereas 3 specimens from 2 cases yielded score +2, but all the 3 specimens with score 2+ subsequently proved negative for Her2/neu gene amplification by fluorescence in situ hybridization. Of 10 primaries studied, 4 tumors showed positive p53 immunoreaction in more than 25% of the cells comprising the malignant portion of the lesions, in 2 cases, a minority of the neoplastic cells (10%-20%) demonstrated nuclear staining, whereas the remaining 4 cases were negative. Of 9 specimens of hidradenocarcinoma studied for TP53 mutations, 2 harbored mutations, whereas the remaining 7 specimens showed the wild-type sequence. Of 11 specimens studied for translocation t(11;19), 2 cases harbored the translocation. It is concluded that cutaneous hidradenocarcinomas show some microscopic heterogeneity and comprise both low- and high-grade lesions that cytologically are similar to their benign counterpart, the hidradenoma. Within the spectrum of low-grade lesions, there seem to exist tumors almost indistinguishable from hidradenomas but still being capable of regional or distant metastasis. Similar to hidradenomas, hidradenocarcinomas show a t(11;19) translocation, but it is a significantly rarer event. Even rarer is the amplification of the Her2/neu gene. Of note is the relatively low frequency of TP53 mutations despite a high rate of p53 protein expression at the immunohistochemical level.
- MeSH
- adenokarcinom chemie genetika patologie MeSH
- amplifikace genu * MeSH
- buněčná diferenciace MeSH
- dospělí MeSH
- hybridizace in situ fluorescenční MeSH
- imunohistochemie MeSH
- lidé středního věku MeSH
- lidé MeSH
- lidské chromozomy, pár 11 * MeSH
- lidské chromozomy, pár 19 * MeSH
- mutace * MeSH
- mutační analýza DNA MeSH
- nádorový supresorový protein p53 analýza genetika MeSH
- nádory potních žláz chemie genetika patologie MeSH
- nekróza MeSH
- proliferace buněk MeSH
- receptor erbB-2 analýza genetika MeSH
- regulace genové exprese u nádorů MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- translokace genetická * MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- ERBB2 protein, human MeSH Prohlížeč
- nádorový supresorový protein p53 MeSH
- receptor erbB-2 MeSH
- TP53 protein, human MeSH Prohlížeč
BACKGROUND: The number and complexity of repetitive elements varies between species, being in general most represented in those with larger genomes. Combining the flow-sorted chromosome arms approach to genome analysis with second generation DNA sequencing technologies provides a unique opportunity to study the repetitive portion of each chromosome, enabling comparisons among them. Additionally, different sequencing approaches may produce different depth of insight to repeatome content and structure. In this work we analyze and characterize the repetitive sequences of Triticum aestivum cv. Chinese Spring homeologous group 4 chromosome arms, obtained through Roche 454 and Illumina sequencing technologies, hereinafter marked by subscripts 454 and I, respectively. Repetitive sequences were identified with the RepeatMasker software using the interspersed repeat database mips-REdat_v9.0p. The input sequences consisted of our 4DS454 and 4DL454 scaffolds and 4ASI, 4ALI, 4BSI, 4BLI, 4DSI and 4DLI contigs, downloaded from the International Wheat Genome Sequencing Consortium (IWGSC). RESULTS: Repetitive sequences content varied from 55% to 63% for all chromosome arm assemblies except for 4DLI, in which the repeat content was 38%. Transposable elements, small RNA, satellites, simple repeats and low complexity sequences were analyzed. SSR frequency was found one per 24 to 27 kb for all chromosome assemblies except 4DLI, where it was three times higher. Dinucleotides and trinucleotides were the most abundant SSR repeat units. (GA)n/(TC)n was the most abundant SSR except for 4DLI where the most frequently identified SSR was (CCG/CGG)n. Retrotransposons followed by DNA transposons were the most highly represented sequence repeats, mainly composed of CACTA/En-Spm and Gypsy superfamilies, respectively. This whole chromosome sequence analysis allowed identification of three new LTR retrotransposon families belonging to the Copia superfamily, one belonging to the Gypsy superfamily and two TRIM retrotransposon families. Their physical distribution in wheat genome was analyzed by fluorescent in situ hybridization (FISH) and one of them, the Carmen retrotransposon, was found specific for centromeric regions of all wheat chromosomes. CONCLUSION: The presented work is the first deep report of wheat repetitive sequences analyzed at the chromosome arm level, revealing the first insight into the repeatome of T. aestivum chromosomes of homeologous group 4.
- MeSH
- chromozomy rostlin genetika MeSH
- DNA rostlinná analýza MeSH
- fyzikální mapování chromozomů MeSH
- pšenice genetika MeSH
- repetitivní sekvence nukleových kyselin * MeSH
- sekvenční analýza DNA metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- DNA rostlinná MeSH
