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× Leymotrigia bergrothii is a presumed hybrid of Leymus arenarius and Elytrigia repens . This article investigates the hybrid origin and genome composition of this species. These plants are sterile, do not undergo pollination, and do not produce seeds; occasionally, underdeveloped stamens containing abortive pollen grains form in individual spikelets. The karyotype analysis of root meristem cells revealed a diploid chromosome number of 49 in × L. bergrothii , reported here for the first time. Subsequently, we examined the intragenomic polymorphism of the transcribed spacer ITS1 in several species of Elytrigia , Elymus , Leymus , Hordeum , and Psathyrostachys , and compared the ribotype patterns of these species with those of × L. bergrothii. It is shown that the St-ribotype variants found in Elytrigia repens and Elytrigia pseudocaesia , as well as the ribotypes of the La family, which dominate in the genome of Leymus arenarius , correspond to major ribotypes in × L. bergrothii . The ribotypes of the St and La families are present in the nuclear genome of × L. bergrothii in almost equal proportions. A comparison of intron and exon sequences of the trn L gene in the chloroplast DNA of L ey mus arenarius , Elytrigia repens , and × L. bergrothii showed that this region in × L. bergrothii is identical or very close to that of Elytrigia repens , suggesting that Elytrigia repens was the cytoplasmic donor to × L. bergrothii. Thus, our study confirms the hypothesis that this species represents a sterile first-generation hybrid of Leymus arenarius and Elytrigia repens , reproducing vegetatively.

The species of the genus Deschampsia are difficult for identification, and the genus is difficult for taxonomic treatment. The regions of 35S rRNA genes were studied for the species of the genus Deschampsia of different geographical origin with a method of sequencing by Sanger (ITS1-5.8S rRNA gene-ITS2, 14 species) and with a method of a locus-specific next-generation sequencing (NGS) on the Illumina platform (ITS1-5.8S rRNA, 7 species). All species of Deschampsia formed one clade; the species, referred by some authors on the basis of morphological characters to the species D. cespitosa s.l., entered one subclade. Subantarctic species formed a separate subclade and their ribotypes formed their own subnetwork. Avenella flexuosa , earlier referred to Deschampsia , entered the other clade, though this species contains some ribotypes common with some Deschampsia species. Deschampsia pamirica and related mountain species have their own specific ribotype groups. On the network of the ribotypes, one can see that D. cespitosa from Great Britain forms a network with some species, but D. cespitosa from the USA forms its own network. Ribotype analysis of each sample revealed traces of introgression with Deyeuxia/Calamagrostis in D. cespitosa and with A. flexuosa and probable introgression of Northern and subantarctic species.

We performed next-generation sequencing of the 18S rDNA-ITS1-5.8S rDNA region along with traditional Sanger sequencing of rbc L, mat K, ndh F, and ITS1-5.8S rDNA-ITS2 to clarify the hybridization pattern in the subtribe Alopecurinae and in the genus Alopecurus in particular. Our data support the hybrid origin of Alopecurus × brachystylus from hybridization between A. geniculatus (sect. Alopecurium ) and A. pratensis (sect. Alopecurus ). Moreover, in the rDNA of hybrid A. × brachystylus , only A. aequalis -like ribotypes from tetraploid A. geniculatus participated. Surprisingly, we found the traces of introgression of A. arundinaceus -like ribotypes not only in hybrid A. × marssonii ( A. geniculatus × A. arundinaceus ) but in A. aequalis s. str. as well. A high-polyploid group from the section Alopecurus , A . aggr. alpinus has undoubted hybrid origin: e. g., A. brachystachyus has rDNA from the sect. Alopecurium . Alopecurus alpinus , with its allies, is clearly distinct from other members of the sect. Alopecurus (especially by maternal line) and thus we can re-establish a previous opinion about the separate group to which A. alpinus belongs. Species from the section Colobachne (presumably Alpine grasses from Ancient Mediterranean region) probably hybridized with the A. alpinus group. Even A. myosuroides (sect. Pseudophalaris ) that could be referred to the separate genus has ribotypes common with the species of the section Alopecurium ( A. aequalis , A. geniculatus ) in one of the accessions. Additionally, we found that the possible polyphyletic origin of the genus Limnas . Limnas stelleri is very close to Alopecurus magellanicus according to NGS data, while L. malyschevii is more or less distinct from other studied species of the genus Alopecurus .

In our article, we analyzed new data on the origin of the hybrid genus × Trisetokoeleria . According to the morphological criteria × T. jurtzevii is a hybrid between Koeleria asiatica s. l. and Trisetum spicatum , × T. taimyrica , and originated from Koeleria asiatica s. l. and Trisetum subalpestre , × T. gorodkowii , a hybrid between Koeleria asiatica and Trisetum ruprechtianum . Later × T. taimyrica was transferred to Koeleria . Parental taxa are prone to active hybridization themselves, thus, new methods of next-generation sequencing (NGS) were needed to clarify the relationships of these genera. For NGS we used the fragment 18S rDNA (part)-ITS1-5.8S rDNA (totally 441 accessions). We analyzed ITS1-5.8S rDNA-ITS2 region, trn L- trn F and trn K- rps 16 from eight samples of the five species, using the Sanger method: × Trisetokoeleria jurtzevii , × T. taimyrica , Koeleria asiatica , Sibirotrisetum sibiricum (= Trisetum sibiricum ), and Trisetum spicatum . We also studied the pollen fertility of × Trisetokoeleria and its possible progenitors. Our data partly contradicted previous assumptions, based on morphological grounds, and showed us a picture of developed introgression within and between Koeleria and Trisetum . × T. jurtzevii , a totally sterile hybrid formed rather recently. We can suppose that × T. jurtzevii is a hybrid between K. asiatica and some Trisetum s. str. Species, but not T. spicatum . × T. gorodkowii , a hybrid in the stage of primary stabilization; it has one unique ribotype related to T. spicatum s. l. The second parental species is unrelated to Trisetum ruprechtianum . × T. taimyrica and is a stabilized hybrid species; it shares major ribotypes with the T. spicatum / T. wrangelense group and has a minor fraction of rDNA related to genus Deyeuxia s. l.

Zingeria (Poaceae) is a small genus that includes Z. biebersteiniana, a diploid species with the lowest chromosome number known in plants (2n = 4) as well as hexaploid Z. kochii and tetraploid Z. pisidica, and/or Z. trichopoda species. The relationship between these species and the other low-chromosomes species Colpodium versicolor are unclear. To explore the intragenomic polymorphism and genome composition of these species we examined the sequences of the internal transcribed spacer 1 of the 35S rRNA gene via NGS approach. Our study revealed six groups of ribotypes in Zingeria species. Their distribution confirmed the allopolyploid nature of Z. kochii, whose probable ancestors were Colpodium versicolor and Z. pisidica. Z. pisidica has 98% of rDNA characteristic only for this species, and about 0.3% of rDNA related to that of Z. biebersteiniana. We assume that hexaploid Z. kochii is either an old allopolyploid or a homodiploid that has lost most of the rRNA genes obtained from Z. biebersteiniana. In Z. trichopoda about 81% of rDNA is related to rDNA of Z. biebersteiniana and 19% of rDNA is derived from Poa diaphora sensu lato. The composition of the ribotypes of the two plants determined by a taxonomy specialist as Z. pisidica and Z. trichopoda is very different. Two singleton species are proposed on this base with ribotypes as discriminative characters. So, in all four studied Zingeria species, even if the morphological difference among the studied species was modest, the genomic constitution was significantly different, which suggests that these are allopolyploids that obtained genomes from different ancestors.

Background: Grasslands in the Arctic tundra undergo irreversible degradation due to climatic changes and also over-exploitation and depletion of scarce resources. Comprehensive investigations of cytogenomic structures of valuable Arctic and sub-Arctic grassland species is essential for clarifying their genetic peculiarities and phylogenetic relationships, and also successful developing new forage grass cultivars with high levels of adaptation, stable productivity and longevity. We performed molecular cytogenetic characterization of insufficiently studied pasture grass species (Poaceae) from related genera representing two neighboring clades: 1) Deschampsia and Holcus; 2) Alopecurus, Arctagrostis and Beckmannia, which are the primary fodder resources in the Arctic tundra., Results: We constructed the integrated schematic maps of distribution of these species in the northern, central and eastern parts of Eurasia based on the currently available data as only scattered data on their occurrence is currently available. The species karyotypes were examined with the use of DAPI-banding, multicolour FISH with 35S rDNA, 5S rDNA and the (GTT) 9 microsatellite motif and also sequential rapid multocolour GISH with genomic DNAs of Deschampsia sukatschewii, Deschampsia flexuosa and Holcus lanatus belonging to one of the studied clades. Cytogenomic structures of the species were specified; peculiarities and common features of their genomes were revealed. Different chromosomal rearrangements were detected in Beckmannia syzigachne, Deschampsia cespitosa and D. flexuosa; B chromosomes with distinct DAPI-bands were observed in karyotypes of D. cespitosa and H. lanatus., Conclusions: The peculiarities of distribution patterns of the examined chromosomal markers and also presence of common homologous DNA repeats in karyotypes of the studies species allowed us to verify their relationships. The obtained unique data on distribution areas and cytogenomic structures of the valuable Arctic and sub-Arctic pasture species are important for further genetic and biotechnological studies and also plant breeding progress.

Region ITS1–5.8S rDNA–ITS2 is sequenced in 27 varieties of cultivated ornamental peonies, ten of which presumably originate from Paeonia lactiflora, one from P. officinalis, 13 from hybridization of P. lactiflora and P. peregrina, or P. officinalis, and three are Itoh hybrids. Comparative analysis of distribution patterns of polymorphic sites (PS) for the obtained DNA sequences and data from GenBank is carried out. Hypotheses of origin of the studied varieties, except for two, which, as previously assumed, originate from hybridization of P. lactiflora and P. peregrina, are confirmed. It is shown that the sequence ITS1–5.8S rDNA–ITS2 is a good genetic marker for cultivars of the P. lactiflora group and Itoh hybrids, and that the PS distribution patterns in these sequences can provide valuable information on the kinship and origin of individual varieties. However, insufficient knowledge of wild species from the P. officinalis kinship group limits the use of this marker in the study of varieties obtained through interspecific hybridization within the Paeonia section.

This paper briefly discusses the mechanisms by which either diploid 2n or basal chromosome number x change during grass karyotype evolution.

The ITS1-5.8S rDNA-ITS2 regions of 33 accessions belonging to 16 species and five natural and garden interspecific hybrids of the genus Paeonia L. were sequenced. Chromatograms of the peony hybrids demonstrated the presence of the signals, corresponding to two different nucleotides at the positions differing in the parents, indicating that in the hybrids, no rDNA isogenization usually occurred, and they preserved rDNA of both parents. Analysis of these polymorphic sites (PS) showed that P. x majkoae was interspecific hybrid between P. tenuifolia and P. caucasica. The ITS of P. hybrida differs from ITS of P. x majkoae in 19 mutations. Because of this, P. x majkoae is definitely not synonymous to P. hybrida. Comparative analysis of ITS 1-5.8S rDNA-ITS2 showed that species diversity in section Paeonia was based on recombination as a result of intraspecific hybridization of three haplotype families. Specifically, haplotypes A, typical of the P. tenuifolia and P. anomala genomes, haplotypes B, typical of P. mlokosewitschii and P. obovata, and haplotypes of family C, currently represented in rDNA of diploid and tetraploid forms of some Caucasian and Mediterranean species. The ITS regions many diploid peonies contain no dimorphic sites, while P. oreogeton, P. cambessedesii, P. rhodia, and P. daurica carry from ten to 17 PS, and supposed to be the interspecific hybrids. Most of the tetraploid peonies contain from six to 18 PS in the ITS regions. These are alloploids with one of the parental genomes similar to that of P. mlokosewitschii (B1), or P. obovata (B3). The second parental genome in P. banatica, P. peregrina, and P. russii is represented by the genome, close to that of P. tenuifolia (A). P. macrophylla, P. mascula, P. coriacea, P. wittmanniana, and P. tomentosa carry genome of series B and genome of series C, which slightly resembles genome A.

The involvement of present-day diploid bluegrass species in the formation of polyploid genomes was investigated using comparison of sequences of internal transcribed spacers ITS1 and ITS2, and the 5.8S rDNA sequence. It was demonstrated that highly polyploid New Zealand bluegrasses, P. cita (2n = 84; ca. 96 to 100), P. chathamica (2n = 112), and P. litorosa (2n = 263 to 266) formed separate highly supported clade together with tetraploids (2n = 28) P. intrusa, P. anceps, and P. trioides (Austrofestuca littoralis). Among the diploid species (2n = 14), the closest relatives of these species, as well as of the polyploid species of section Poa, are the genomes of Eurasian species P. remota, P. chaixcii (sect. Homalopoa), P densa (Bolbophorum), and P. sibirica (sect. Macropoa). Nuclear genomes of polyploid Stenopoa, Tichopoa, Oreinos, and Secundae are definitely related to the genome of Arctic species P. pseudabbreviata (sect. Abbreviatae). On the contrary, judging by the genes for nuclear 45S rRNA, genomes of diploid P. trivialis (sect. Pandemos), P. annua, and P. supina (sect. Ochlopoa both) are only remotely related to the genomes of highly polyploid species (distances p between them and other bluegrass species from different sections of subgenus Poa constitute 6-10% and 11-15%, respectively). The conclusion on the relationships between highly polyploid and diploid bluegrass species was tested using analysis of synapomorphic mutations in the 5.8S rRNA gene. It was demonstrated that genomes of Poa eminens (2n = 42) and P. schischkinii (2n = 70) (sect. Arctopoa both) were noticeably different in ITS regions from the genomes of the members of the type subgenus Poa. A comparison of the Arctopoa ITS regions showed that the differences between them constituted only 0.2%. At the same time, p distances between the Arctopoa ITS and those from the species belonging to other sections of the genus Poa varied from 5 to 14%. South American species P chonotica (sect. Andinae) (=Ncoraepoa chonotica) (2n = 42) was found to be related to Arctagrostis, Festucella, and Hookerochloa, being at the same time quite distant from the other species of the genus Poa. Polymorphic in chromosome number highly polyploid species of Northern Hemisphere, P. arctica (2n = 42 to 106), P. turneri (2n = 42, 63 to 64), and P. smirnovii (2n = 42, 70) (sect. Malacanthae) are relative to a large group of tetraploid (2n = 28) endemic bluegrass species from New Zealand and sub-Antarctic islands (P. novae-zelandiae and allied species).

Chromosome C-banding and two-color fluorescent in situ hybridization (FISH) were used to compare the chromosomes, to identify the chromosomal localization of the 45S and 5S rRNA genes, and to analyze the sequences of internal transcribed spacers 1 and 2 (ITS1 and ITS2) of the 45S rRNA genes in the genomes of grasses Zingeria biebersteiniana (2n = 4), Z. pisidica, Z. trichopoda (2n = 8), Colpodium versicolor (2n = 4), and Catabrosella variegata (syn. Colpodium variegatum) (2n = 10). Differences in C-banding pattern were observed for two Z. biebersteiniana accessions from different localities. Similar C-banding patterns of chromosomes 1 and 2 were demonstrated for the Z. pisidica and Z. biebersteininana karyotypes. Chromosome C banding and localization of the 45S and 5S rRNA genes on the chromosomes of the two Zingeria species confirmed the assumption that Z. pisidica is an allotetraploid with one of the subgenomes similar to the Z. biebersteiniana genome. ITS comparisons showed that the unique two-chromosome grasses (x = 2)-Z. biebersteiniana (2n = 4), Z. trichopoda (2n = 8), Z. pisidica (2n = 8), and C. versicolor (2n = 4), which were earlier assigned to different tribes of subtribes of the family Poaceae-represent two closely related genera, the genetic distance (p-distance) between their ITSs being only 1.2-4.4%. The Zingeria species and C. versicolor formed a common clade with Catabrosella araratica (2n = 42, x = 7) on a molecular phylogenetic tree. Thus, the karyotypes of Zingeria and Colpodium, which have the lowest known basic chromosome number (x = 2), proved to be monophyletic, rather than originating from different phylogenetic lineages.

Variations of nucleotide composition and frequency of CpG and CpNpG sequences in the clusters of nuclear ribosomal genes of taxa, belonging to two phylogenetic branches of Angiospermae have been statistically analyzed. This region of eucaryotic genomes is nucleolus organizer and functions in a separate compartment of cell nucleus that can do running here processes it is enough specific. It is shown that level of evolution advance of a taxon, defined on morphological data, is in positive correlation with quantitative value of dC, CpG and CpNpG. This is found in contradiction with beliefs about the general rules of the transformation of nucleotide composition in evolution, that suggest a CpG suppression. Cryptaffinous taxa being connecting link between a well morphological outlined taxonomic group, differ to be raised in

Chromosome banding with nucleotide base-specific fluorochromes chromomycin A3 (CMA) and Hoechst 33258 (H33258) was used to study the karyotypes and to construct cytological maps for diploid Trillium camschatcense (2n = 10), tetraploid T. tschonoskii (2n = 20), hexaploid T. rhombifolium (2n = 30), and a triploid T. camschatcense x T. tschonoskii hybrid (T. x hagae, 2n = 15). With H33258, species- and genome-specific patterns with numerous AT-rich heterochromatin bands were obtained for each of the four forms; CMA revealed a few small, mostly telomeric GC-rich bands. In T. tschonoskii, the two subgenomes were similar to each other and differed from the T. camschatcense genome; on this evidence, the species was considered to be a segmental allotetraploid. In T. x hagae, one T. camschatcense and both T. tschonoskii subgenomes were identified. The subgenomes of T. rhombifolium only partly corresponded to the T. camschatcense and T. tschonoskii genomes, in contrast to the morphologically identical Japanese species T. hagae. This was assumed to indicate that allohexaploids T. rhombifolium and T. hagae originated independently at different times; i.e., their origin is polyphyletic. Based on the chromosome maps, a new nomenclature was proposed for the Trillium genomes examined: K1K1 for T. camschatcense, T1T1T2T2 for T. tschonoskii, T1T1T2T2 for T. x hagae, and K1RK1RT1RT1RT2RT2R for T. rhombifolium.

Chromosome banding patterns of Allium cepa L. were obtained by using fluorochrome combinations chromomycin A3 (CMA) + 4',6-diamidino-2-phenylindole (DAPI), DAPI + actinomycin D (AMD) and propidium iodide (PI) + DAPI. In A. cepa, telomeric heterochromatin displayed dull fluorescence after staining with DAPI and DAPI/AMD. After staining with the GC-specific CMA and AT-specific DAPI, the CMA-positive fluorescence of the NOR region and the telomeric bands of C-heterochromatin was observed. In combination with DAPI, PI, a dye with low AT/GC specificity, produced almost uniform fluorescence of chromosomal arms and heterochromatin, whereas the NOR-adjoining regions displayed bright fluorescence. Denaturation of chromosomal DNA (95 degrees C for 1-3 min) followed by renaturation in the 2 x SSC buffer (37 degrees C, 12 h) altered the chromosome fluorescence patterns: specific PI-positive bands appeared and the contrast of CMA-banding increased. Bright fluorescence of the NOR and adjoining regions was also observed in the case. Three-minute denaturation led also to a bright PI-positive fluorescence of telomeric heterochromatin. The denaturation of chromosomal DNA before staining results in changes of the DAPI fluorescence pattern and in the appearance of DAPI fluorescence in GR-rich NOP regions. The mechanisms underlying the effects of denaturation/renaturation procedures on chromosome banding patterns obtained with different fluorochromes are discussed.

Cold-induced decondensation of heterochromatic regions (CSR-bands) in Paris hainanensis (= Daiswa hainanensis Merrill Takht.) (2n = 10; 10 + b) was studied. The comparison of CSR-banding patterns with those obtained by nucleotide-specific staining with fluorochromes DAPI and chromomycin A3 demonstrated that low temperatures induced decondensation only of large AT-rich heterochromatic regions. It is suggested that this is characteristic of all plant species.

The karyotype of the chicken Gallus gallus domesticus was studied by means of pulsed-field electrophoresis. An electrokaryogram was obtained for the microchromosomal (MI) portion of the chicken genome. Chicken MIs were separated into two fractions. A fraction with a higher mobility included MIs sized 3.4-4.8 Mb; the lower size limit of a less mobile fraction corresponded to MIs of approximately 5 Mb. The smallest MI in the chicken karyotype was estimated at 3.4 +/- 0.25 Mb.

Using nucleotide-specific agents Hoechst 33258, actinomycin D, chromomycin A3, and distamycin A, the Paris quadrifolia L. karyotype, and the location and nucleotide composition of heterochromatic bands were studied. The chromosome ideogram of H33258/AMD and CMA/DA heterochromatic bands was created by an image analysis system with the Videotest-Kario software. By fluorescence in situ hybridization, the 18S and 26S rRNA genes were mapped.