Karyological features and cytotaxonomy of the tribe Vernonieae (Asteraceae)

Chromosome numbers are reported for 29 populations of 19 Vernonieae taxa collected mainly in the northeastern region of Brazil. Among them, data for five genera (Blanchetia, Rolandra, Pithecoseris, Stilpnopappus and Vanillosmopsis) are here reported for the first time, and the first chromosome counts are presented for 12 species. Chromosome numbers are quite diverse among and sometimes within genera, especially in the controversial and large subtribe Vernoniinae. The numbers varied from 2n = 18 to 2n = ~72. The main karyoevolutionary mechanism seems to be dysploidy, while polyploidy is probably associated with ancient hybridization processes generating most paleotetraploid genera. All studied species presented semi-reticulated interphase nuclei and proximal-early condensing behavior in prophase to prometaphase. In one species (Vernonia condensata with 2n = 40) fluorochrome staining with CMA/DAPI revealed five chromosome pairs bearing subterminal CMA⁺/DAPI^? heterochromatin, probably NOR-associated, revealing the existence of low amounts of satellite DNA. The role of these features in the evolution of the tribe is discussed, revealing some interesting aspects for understanding of the Vernonieae karyoevolution, especially regarding neotropical members.     

Chromosomal evolution in Pleurothallidinae (Orchidaceae: Epidendroideae) with an emphasis on the genus Acianthera: chromosome numbers and heterochromatin

In this study, we analysed chromosome number variation and chromomycin A₃/4′,6‐diamidino‐2‐phenylindole (CMA/DAPI) banding patterns in 48 species belonging to 12 genera of subtribe Pleurothallidinae (Orchidaceae) in order to understand the chromosome evolution based on recent phylogenetic hypotheses and taxonomic treatments. All species had small chromosomes, with numbers ranging from 2n = 20 in two Specklinia spp. to 2n = 80 in an unidentified Octomeria sp. In Acianthera, the most highly represented genus in this study, a great diversity of chromosome number and pattern of fluorescent bands was observed, showing heterochromatin accumulation in Acianthera section Sicariae subsection Pectinatae. Interspecific ascending and, mainly, descending dysploidy were the main mechanisms of chromosome number evolution in subtribe Pleurothallidinae. For Pleurothallidinae, x = 20 is suggested as the basic chromosome number, the same suggested for the related subtribe Laeliinae and for the whole tribe Epidendreae. The Brazilian species of the mega‐genus Stelis had chromosomes with small amounts of heterochromatin and chromosome numbers based on x₂ = 16. These are generally divergent from those reported for Andean and Meso‐American species, but in agreement with the monophyletic hypothesis proposed for Stelis spp. with a Brazilian Atlantic distribution. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 178 , 102–120.     

Chromosome Numbers and Karyotypes of Species of Vernonia sect. Lepidaploa (Asteraceae: Vernonieae)

Vernonia is the largest genus of the tribe Vernonieae (Asteraceae) and comprises more than 1,000 species. In the present study we explore chromosome number and karyotype variation of eight species treated within different subsections of the section Vernonia sect. Lepidaploa. We aimed to explore if these data support the recognition of a single large genus (sensu Baker) or favor its splitting into 22 small genera (sensu Robinson). The species were collected in ??cerrado??, rupicolous and disturbed areas in the states of S?o Paulo and Minas Gerais, Brazil. Chromosome numbers varied from 2n?=?32 to 60. Most chromosomes were small, and the karyotype analysis revealed a predominance of metacentric and some submetacentric chromosomes. The karyotype symmetry in Vernonia was moderate (TF% 32.2 to 45.9), with the most symmetrical karyotype observed in V. rubriramea. The results obtained here did not conclusively support any of the taxonomic proposals for Vernonia due to the absence of distinctive or characteristic karyotype patterns for any of the taxonomic groupings, i.e., sections and subsections (sensu Baker) or new genera (sensu Robinson). Nevertheless, a tenuous relationship was observed between the chromosome numbers reported in the literature, those recorded here, and the taxonomic alterations suggested by Robinson for the genera Lessingianthus, Chrysolaena, and Vernonanthura that were originally part of Vernonia sensu Baker.     

Karyotype analysis and DNA content in some species of Chrysolaena (Vernonieae,Asteraceae)

Cytogenetic characterization by karyotyping and determination of DNA content by flow cytometry of five species of Chrysolaena (Vernonieae, Asteraceae) was performed. This is the first study of nuclear DNA content realized in the genus. The 2C-values were compared with the ploidy level and the total karyotype length (TKL) of each species. Mitotic analysis revealed a base chromosome number x = 10 for all entities and different ploidy levels, from diploid (2n = 2x = 20) to octoploid (2n = 8x = 80). All species showed bimodal karyotypes composed of metacentric and submetacentric chromosomes. The average chromosome size (ML) varied from 1.86 μm to 2.70 μm, while the TKL ranged from 18.65 μm to 80.55 μm. The intrachromosomal asymmetry index (A₁) varied from 0.27 to 0.38, while the interchromosomal asymmetry index (A₂) ranged from 0.19 to 0.25. A new cytotype is reported for the first time for C. propinqua. Accessory chromosomes found in C. verbascifolia, C. cognata, C. flexuosa, and C. propinqua are also reported as new.     

DNA content in species of Vernonia and Vernonanthura from South America: An approach to systematics and evolution of the Vernonieae (Asteraceae)

There are relatively few studies of DNA content in the Vernonieae (Asteraceae) tribe. The first studies were realized in the Lessingianthus genus and determined the DNA content of 25 species. After DNA content, ploidy level and the total karyotype were compared in 6 Chrysolaena species. The aim of this study was to present, for the first time, the DNA content values of Vernonanthura and Vernonia and to thereby expand knowledge of the Vernonieae tribe. A total of 19 natural populations belonging to the genera Vernonanthura and Vernonia were studied for the first time. The results were compared with other Vernonieae genera and with other Asteraceae tribes. Our results found that Vernonieae have the smallest range of 1C value variation in Asteraceae. Furthermore, there were differences in the DNA content of Vernonia and Vernonanthura. These results show that low DNA content and herbaceous habit in Vernonia are characters derived from the higher DNA content and woody habit present in Vernonanthura. These results could indicate a hybrid origin of one species and allow the determination of both the ploidy and chromosome number of other taxa. The results observed in Vernonanthura species showed a highly significant correlation between 1C-value and latitude.     

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.     

Karyotype analysis and heterochromatin differentiation with Giemsa C-banding and fluorescent counterstaining inCephalanthera (Orchidaceae)

Detailed studies of the chromosomes of the three Austrian species of the genusCephalanthera showed them all to have basically similar karyotypes. BothC. damasonium (2n = 36) andC. longifolia (2n = 32) have three large and several classes of smaller chromosome pairs. The karyotype ofC. rubra (2n = 44) is composed of four large and several groups of smaller pairs. The heterochromatin in these species amounts to about 10% of total karyotype length. All the chromosomes have Giemsa-positive centromeres, but only a few have intercalary or terminal bands. Using differential fluorescent staining with DAPI/actinomycin D, quinacrine/actinomycin D (both A-T specific), and chromomycin A₃/distamycin A (G-C specific) three different types of major heterochromatic bands can be characterized in respect of their satellite DNA composition: highly A-T rich, slightly A-T rich, and very G-C rich. The chromosomes ofC. longifolia contain more A-T rich C-bands than those ofC. damasonium, while the latter's have more G-C rich heterochromatin. In both species several C-bands appear as secondary constrictions or gaps in the Feulgen-stained chromosomes, but most likely, in each species there is only one pair of chromosomes where the secondary constrictions function as nucleolus organizing regions. No major intraspecific variation could be observed except on one small chromosome pair ofC. longifolia which had a heteromorphic C-band in most individuals. Possible pathways of karyotype evolution involving polyploidy and Robertsonian events are discussed.     

Chromosome evolution in the Brachytheciaceae

Abstract

Twenty-eight moss species have been investigated cytologically. Mitotic karyograms have been presented for most of these species and these show a remarkable similarity in the n = 11 karyotype. Nine of the eleven chromosomes have terminal or sub-terminal centromeres. Lightly staining heterochromatic bands frequently occur along the axis of the chromosomes and very often these are the sites of chromosome bending; it has been suggested that these may be areas of neocentric activity. The karyotypes investigated show a reduction series of chromosome number which is paralleled by an increase in the number of long metacentrics; since the number of major chromosome arms is maintained in most of the species it has been suggested that Robertsonian fusion has been an important mechanism in the evolution of the Bracytheciaceae, and probably in all Diplolepidous mosses. Polyploidy has also played an important role in speciation. Finally it has been proposed that n = 11 is the primary basic number in the Diplolepideae.     

Distribution and Evolution of Repeated Sequences in Genomes of Triatominae (Hemiptera-Reduviidae) Inferred from Genomic In Situ Hybridization

The subfamily Triatominae, vectors of Chagas disease, comprises 140 species characterized by a highly homogeneous chromosome number. We analyzed the chromosomal distribution and evolution of repeated sequences in Triatominae genomes by Genomic in situ Hybridization using Triatoma delpontei and Triatoma infestans genomic DNAs as probes. Hybridizations were performed on their own chromosomes and on nine species included in six genera from the two main tribes: Triatomini and Rhodniini. Genomic probes clearly generate two different hybridization patterns, dispersed or accumulated in specific regions or chromosomes. The three used probes generate the same hybridization pattern in each species. However, these patterns are species-specific. In closely related species, the probes strongly hybridized in the autosomal heterochromatic regions, resembling C-banding and DAPI patterns. However, in more distant species these co-localizations are not observed. The heterochromatic Y chromosome is constituted by highly repeated sequences, which is conserved among 10 species of Triatomini tribe suggesting be an ancestral character for this group. However, the Y chromosome in Rhodniini tribe is markedly different, supporting the early evolutionary dichotomy between both tribes. In some species, sex chromosomes and autosomes shared repeated sequences, suggesting meiotic chromatin exchanges among these heterologous chromosomes. Our GISH analyses enabled us to acquire not only reliable information about autosomal repeated sequences distribution but also an insight into sex chromosome evolution in Triatominae. Furthermore, the differentiation obtained by GISH might be a valuable marker to establish phylogenetic relationships and to test the controversial origin of the Triatominae subfamily.     

Chromatin differentiation between Theobroma cacao L. and T. grandiflorum Schum

A comparative analysis of mitotic chromosomes of Theobroma cacao (cacao) and T. grandiflorum (cupuaçu) was performed aiming to identify cytological differences between the two most important species of this genus. Both species have symmetric karyotypes, with 2n = 20 metacentric chromosomes ranging in size from 2.00 to 1.19 μm (cacao) and from 2.21 to 1.15 μm (cupuaçu). The interphase nuclei of both species were of the arreticulate type, displaying up to 20 chromocentres, which were more regularly shaped in cacao than in cupuaçu. Prophase chromosomes of both species were more condensed in the proximal region, sometimes including the whole short arm. Both species exhibited only one pair of terminal heterochromatic bands, positively stained with chromomycin A ₃ , which co-localized with the single 45S rDNA site. Each karyotype displayed a single 5S rDNA site in the proximal region of another chromosome pair. Heterochromatic bands were also observed on the centromeric/pericentromeric regions of all 20 chromosomes of cacao after C-banding followed by Giemsa or DAPI staining, whereas in cupuaçu they were never detected. These data suggest that the chromosomes of both species have been largely conserved and their pericentromeric chromatin is the only citologically differentiated region.     

Cytogenetics of Alismatales s.s.: chromosomal evolution and C-banding

Ten species of Alismatales, five of Alismataceae, four of Limnocharitaceae and one of Hydrocharitaceae were studied with regard to chromosome number, chromosome morphology, and pattern of Giemsa C-bands. The genus Echinodorus had a diploid chromosome number of 22 for all species that were analyzed and a karyotypic formula of 2m + 20a. For the family Limnocharitaceae, Hydrocleys nymphoides had a diploid chromosome number of 16, Hydrocleys martii (4m + 2sm + 10a) had a diploid chromosome number of 16, Limnocharis flava had a diploid chromosome number of 20 and L. laforestii (4m + 16a) had a diploid chromosome number of 20. The only species of Hydrocharitaceae that was studied exhibited a karyotype that consisted of a diploid chromosome number of 28 and a karyotypic formula of 4m + 6sm + 4a. The distribution pattern of the C-banded karyotype in Echinodorus showed four blocks of constitutive heterochromatin in two smaller acrocentric pairs that corresponded to the heterochromatic NORs. In E. lanceolatus, 14 bands in the termini of the arms beyond the heterochromatic NORs of seven acrocentric pairs were also observed. Idiograms are presented and the karyotypic evolution patterns for the studied groups are discussed.     

Cytogenetics of Alismataceae and Limnocharitaceae: CMA/DAPI banding and 45S rDNA sites

Species belonging to the Alismataceae (Echinodorus) and Limnocharitaceae (Hydrocleys and Limnocharis) families were analysed by banding with CMA/DAPI fluorochromes, C/CMA/DAPI banding, and in situ hybridization (FISH) with probes that recognise 45S rDNA. All species of Echinodorus presented 2n = 22, but only in E. lanceolatus were DAPI+ telomeric bands in seven chromosome pairs observed. A bimodal karyotype and GC-rich heterochromatin preferably located in two smaller acrocentric pairs that generally corresponded to the number of sites of 45S rDNA. A similar pattern of bands was observed in both Limnocharis species (2n = 20), but the two differed with respect to 45S rDNA, with L. laforestii showing only two sites. Hydrocleys nymphoides and H. martii had a chromosome number of 2n = 16, but the position of the GC-rich heterochromatin associated with the satellite differed among chromosomal types. In this work, the cytotaxonomic implications of these patterns are discussed and correlated with previous data from the literature.     

Analysis of heterochromatin by combination of C-banding and CMA3 and DAPI staining in two fish species (Pimelodidae,Siluriformes)

The chromosomes of Steindachneridion sp. (2n = 56) and Rhamdia quelen (2n = 58) were analyzed by C-banding (CB) and Chromomycin A₃ (CMA₃) and 4,6-diamidino-2-phenylindole (DAPI) staining, separately and consecutively, in order to understand the role of base-specific fluorochrome treatment after CB. Both species' chromosomes shared common staining profiles as follows. CB with Giemsa (CBG) revealed weak heterochromatic blocks in the telomeric regions of some chromosomes and conspicuous bands on the short arms of one chromosome pair, where nucleolar organizer regions (NORs) were evidenced by silver-staining. Without CB pretreatment, the NORs were stained conspicuously with CMA₃, but not with DAPI. The latter uniformly stained all chromosomes, but leaving the NORs pale. Combination of CMA₃ or DAPI staining with CB showed distinctive fluorescent blocks in the NOR-bearing short arms of the single chromosome pair along with several bright fluorescent signals on other chromosomes, which were not evidenced by single CMA₃ or DAPI staining. These results suggest a modification of chromatin structure by CB treatment, which may increase the stainability of CMA₃ and DAPI.     

Comparative cytogenetic analysis of Cestrum (Solanaceae) reveals different trends in heterochromatin and rDNA sites distribution

Species of Cestrum L. (Solanaceae) exhibit large variability in the accumulation of repetitive DNA, although their species possess a stable diploid number with 2n = 16. In this study, we used chromosome banding and fluorescence in situ hybridization (FISH) to characterize the karyotypes and populations of two species, Cestrum nocturnum L. and C. mariquitense Kunth. We also performed a karyotype comparison using 16 idiograms, of which 4 were developed in this study and 12 were obtained from the literature. Cestrum nocturnum displayed more bands than C. mariquitense, but the latter exhibited greater interpopulational variation in the band patterns. There was a tendency for large bands to be located at intercalary/terminal regions and for small bands to be located at intermediate/proximal regions. The idiogram comparison revealed a large variation in the amount, distribution, and size of heterochromatic bands. FISH with rDNA probes revealed stability in the number and location of 5S sites, while 45S was more variable in size and number of sites. Although 45S rDNA always appeared in the subterminal regions, this DNA family exhibited a mobility among chromosome pairs. These data highlight the dynamic of repetitive DNA families in these genomes, as well as the contribution for intra- and interspecific karyotype differentiation in Cestrum.     

Evolutionary trends in the chromosome numbers of swarm-founding social wasps

Chromosome number and morphology are relevant aspects of genomic organization of eukaryotes and are considered key components to comprehend evolutionary mechanisms and karyotypic differentiation. Social wasps belonging to the tribe Epiponini show complex social characteristics that make these insects interesting models for genetic and evolutionary studies. However, there is a paucity of genetic information in social wasps as a whole. Aiming to investigate the process of chromosomal evolution of the social Epiponini wasps, chromosomes of ten species of this group were analyzed using Giemsa and fluorochrome staining techniques and fluorescence in situ hybridization in two species. In this study a high variation in the chromosome number and morphology was found and the previous range of chromosome number of Epiponini was broadened from n = 8–32 to n = 5–33. We also suggest that chromosomal segments with high GC content must have had a key role in the karyotype diversification of these wasps. Moreover, based on a phylogenetic background we find evidence of a main role of chromosomal fusion in the occurrence of gradual decrease of chromosome number in Epiponini during its chromosomal evolutionary history.     

The chromosomes of Lepturinae (Coleoptera: Cerambycidae) II. A study of eight more species,with focus on Desmocerus palliatus

Following the study of 28 species of Lepturinae (Coleoptera: Cerambycidae) the karyotypes of seven additional Palaearctic and one Nearctic species are established. The 19,X male karyotypes found in genera Stictoleptura (four species), Vadonia and Judolia (one species each) confirm the loss of Y chromosome in Lepturini. The 22,XY male karyotype of Cortodera humeralis is very close to that of some species of Rhagiini (genera Gaurotes, Acmaeops, Dinoptera, all 22,XY) and Grammoptera ruficornis (24,XY) recently reported. We propose that these taxa could form a monophyletic group within Rhagiini. The karyotype of the Nearctic species Desmocerus palliates (23,neoXneoXneoY) is quite different and characterized by the presence of many acrocentric chromosomes and a complex autosome–gonosome translocation. Its particular karyotype is compatible with its present classification within a separate tribe, the Desmocerini.     

Genome structure and apomixis in Phoenicaulis (Brassicaceae; Boechereae)

Apomixis in crucifer (Brassicaceae) species is rare, reported in just four genera (Boechera, Draba, Erysimum, and Parrya), and one intergeneric hybrid (Raphanobrassica). It is well studied only in Boechera, where it is widespread among 100+ recognized species. However, its occurrence in related genera of the tribe Boechereae has not been documented previously. Here we analyzed genome evolution, mode of reproduction, and fertility of the monospecific Boechereae genus Phoenicaulis (P. cheiranthoides), endemic to the northwestern United States. We discovered that the species encompasses diploid (2n = 2x = 14), triploid (2n = 3x = 21), and tetraploid (2n = 4x = 28) populations. Comparative chromosome painting analyses revealed that the three karyotypes are essentially structurally identical, differing only in the presence of a largely heterochromatic chromosome (Het) in the triploid and tetraploid cytotypes. The genome structure of Phoenicaulis appeared identical to that of Boechera species previously analyzed, suggesting genomic stasis despite the morphological and molecular divergence of the two genera. This genome colinearity extended to the presence and structure of the Het chromosomes, which are closely associated with apomictic reproduction in Boechera. Interestingly, all three cytotypes of Phoenicaulis proved to be apomictic, regardless of the presence or absence of a Het chromosome, and sexual populations have yet to be identified.     

Cytogenetic characterization of the ant Trachymyrmex fuscus Emery, 1934 (Formicidae: Myrmicinae: Attini) with the description of a chromosomal polymorphism

The genus Trachymyrmex is a key group in the tribe Attini because of its close phylogenetic relationship to leaf-cutter ants, Acromyrmex and Atta. Cytogenetic data are only available for five taxa of Trachymyrmex, with chromosome numbers of 2n = 12, 18, 20 and 22, and morphology with predominantly metacentric chromosomes. The aim of the present study was to characterize the karyotype of the ant Trachymyrmex fuscus Emery, 1934, by means of the number and morphology of its chromosomes, heterochromatin pattern, CMA₃ and DAPI fluorochromes in the population of two nests collected at Paraopeba, state of Minas Gerais, Brazil. Nineteen females presented 2n = 18 chromosomes (16m + 2sm) and a single male presented n = 9 (8m + 1sm). A size chromosomal polymorphism involving the short arm of the submetacentric pair was confirmed by statistical analysis, with three character conditions: heterozygous SB (with a difference in size between the short arms), standard SS (smaller short arms) and homozygote BB (bigger short arms). In the first nest, both SB and SS workers were observed. The other nest contained heterozygous (SB), homozygous (BB), and a male carrying the B chromosome (larger size). The presence of heterochromatin on all centromeric and pericentromeric chromosomes of T. fuscus suggests that the size difference observed in the submetacentric pair in the SB and BB workers is not related to the heterochromatin but to a duplication of euchromatic regions through intra- or inter- chromosomal rearrangements. The fluorochrome CMA₃ matched the C-banding markings, indicating that the heterochromatin is rich in GC base pairs. As far as we know, this is the first chromosomal polymorphism reported in the tribe Attini.     

Chromosome studies in Orchidaceae: karyotype divergence in Neotropical genera in subtribe Maxillariinae

Here, we study karyotype divergence in the closely related genera Brasiliorchis, Christensonella and Trigonidium belonging to subtribe Maxillariinae of subfamily Epidendroideae (Orchidaceae). We compare karyotypes in 15 species by (1) measuring 1C genome sizes, (2) mapping the distribution of 4′,6‐diamidino‐2‐phenylindole and chromomycin A3 chromosome bands and (3) localizing 5S and 45S nuclear ribosomal DNA (rDNA) sequences using fluorescent in situ hybridization. Recently, phylogenetic studies have been conducted to resolve species and genera relationships in subtribe Maxillariinae. We used these phylogenetic trees to map the cytogenetic characters in an evolutionary framework. This has enabled a better understanding of the patterns of genomic divergence in the group. Genome sizes range from 1C = 1.85 to 4.1 pg. The largest, B. schunkeana, shows evidence of genome upsizing, probably through the acquisition of tandem repeats that now form large 4′,6‐diamidino‐2‐phenylindole‐positive blocks of heterochromatin. Our cytogenetic data are consistent with a base chromosome number of 2n = 40, although Christensonella is characterized by a dysploid reduction in chromosome number to 2n = 36. The number of 5S and 45S rDNA sites is variable between species, consistent with high rates of karyotype divergence. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 170 , 29–39.     

Karyotype of Mesembryanthemum crystallinum (Aizoaceae) studied by chromosome banding,FISH with rDNA probes and immunofluorescence detection of DNA methylation

The karyotype of the common ice plant Mesembryanthemum crystallinum L. (Aizoaceae) was studied using Chromomycin A₃ (CMA)/4′,6-diamidino-2-phenylindole (DAPI) staining, fluorescence in situ hybridization with 5S and 18S–5.8S–25S rDNA probes, DAPI/C-banding and immunodetection of 5-methylcytosine. A single bright CMA-band was revealed on the satellite chromosome, whose location was coincided with a position of a site of 18S–5.8S–25S rRNA genes. A site of 5S rRNA genes was observed on one of the other chromosomes. Relatively large DAPI/C-bands were mainly localized in the pericentromeric regions of the chromosomes. DAPI/C-banding patterns allowed us to identify all the chromosomes in the karyotype of M. crystallinum. The methylation of euchromatic chromosome regions was weaker as compared with heterochromatic DAPI/C-bands, which were hypermethylated. The obtained results may provide opportunities for investigating, at the chromosomal level, the genomic changes occurring in M. crystallinum either under salinization or under the action of other stress factors.