Heterochromatin Amplification Research Articles

Chromosomal rearrangements played an important role in the speciation of rice rats of genus Cerradomys (Rodentia, Sigmodontinae, Oryzomyini)

Rodents of the genus Cerradomys belong to tribe Oryzomyini, one of the most diverse and speciose groups in Sigmodontinae (Rodentia, Cricetidae). The speciation process in Cerradomys is associated with chromosomal rearrangements and biogeographic dynamics in South America during the Pleistocene era. As the morphological, molecular and karyotypic aspects of Myomorpha rodents do not evolve at the same rate, we strategically employed karyotypic characters for the construction of chromosomal phylogeny to investigate whether phylogenetic relationships using chromosomal data corroborate the radiation of Cerradomys taxa recovered by molecular phylogeny. Comparative chromosome painting using Hylaeamys megacephalus (HME) whole chromosome probes in C. langguthi (CLA), Cerradomys scotii (CSC), C. subflavus (CSU) and C. vivoi (CVI) shows that karyotypic variability is due to 16 fusion events, 2 fission events, 10 pericentric inversions and 1 centromeric repositioning, plus amplification of constitutive heterochromatin in the short arms of the X chromosomes of CSC and CLA. The chromosomal phylogeny obtained by Maximum Parsimony analysis retrieved Cerradomys as a monophyletic group with 97% support (bootstrap), with CSC as the sister to the other species, followed by a ramification into two clades (69% of branch support), the first comprising CLA and the other branch including CVI and CSU. We integrated the chromosome painting analysis of Eumuroida rodents investigated by HME and Mus musculus (MMU) probes and identified several syntenic blocks shared among representatives of Cricetidae and Muridae. The Cerradomys genus underwent an extensive karyotypic evolutionary process, with multiple rearrangements that shaped extant karyotypes. The chromosomal phylogeny corroborates the phylogenetic relationships proposed by molecular analysis and indicates that karyotypic diversity is associated with species radiation. Three syntenic blocks were identified as part of the ancestral Eumuroida karyotype (AEK): MMU 7/19 (AEK 1), MMU 14 (AEK 10) and MMU 12 (AEK 11). Besides, MMU 5/10 (HME 18/2/24) and MMU 8/13 (HME 22/5/11) should be considered as signatures for Cricetidae, while MMU 5/9/14, 5/7/19, 5 and 8/17 for Sigmodontinae.

H3.1K27me1 loss confers Arabidopsis resistance to Geminivirus by sequestering DNA repair proteins onto host genome

The H3 methyltransferases ATXR5 and ATXR6 deposit H3.1K27me1 to heterochromatin to prevent genomic instability and transposon re-activation. Here, we report that atxr5 atxr6 mutants display robust resistance to Geminivirus. The viral resistance is correlated with activation of DNA repair pathways, but not with transposon re-activation or heterochromatin amplification. We identify RAD51 and RPA1A as partners of virus-encoded Rep protein. The two DNA repair proteins show increased binding to heterochromatic regions and defense-related genes in atxr5 atxr6 vs wild-type plants. Consequently, the proteins have reduced binding to viral DNA in the mutant, thus hampering viral amplification. Additionally, RAD51 recruitment to the host genome arise via BRCA1, HOP2, and CYCB1;1, and this recruitment is essential for viral resistance in atxr5 atxr6. Thus, Geminiviruses adapt to healthy plants by hijacking DNA repair pathways, whereas the unstable genome, triggered by reduced H3.1K27me1, could retain DNA repairing proteins to suppress viral amplification in atxr5 atxr6.

Chromosomal Evolution of the Talpinae.

In recent years, the number of mole species with species status confirmed by genetic methods has been continuously increasing. Unfortunately, cytogenetic data are not yet available for all species. Here, for the first time, a GTG-banded karyotype of the small-toothed mole from Vietnam, Euroscaptor parvidens, a representative of the Eastern clade of the genus Euroscaptor, has been described. Through comparative analysis of available Euroscaptor (Euroscaptor parvidens, Euroscaptor klossi, and Euroscaptor malayana) and Oreoscaptor (Oreoscaptor mizura) karyotypes, we found cytogenetic signatures for each of the studied species. Zoo-FISH with sorted chromosomes of the Siberian mole (Talpa altaica) on chromosome sets of the small-toothed mole (E. parvidens), the small Japanese mole (Mogera imaizumii) from the closely related genus, and the Japanese shrew mole (Urotrichus talpoides) from the tribe Urotrichini made it possible to identify syntenic regions between these species. We propose a possible ancestral karyotype of the tribe and, based on it, traced the features of chromosomal rearrangements accompanying the divergence of moles. The low rates of chromosomal evolution within the species of the genus Talpa-T. altaica and T. europaea-and the high rates of karyotypic reshuffling within the Asian genera of the tribe were confirmed. The karyotype of the Japanese mountain mole O. mizura seems to be the most conserved among the Asian moles. The most frequently occurring types of chromosomal rearrangements in moles are the pericentric inversions and amplification of heterochromatin. The pericentric inversions on four pairs of autosomes are shared between the closely related genera Euroscaptor, Oreoscaptor, and Mogera, while many more apomorphic rearrangements have occurred in each lineage additionally. The highest rate of chromosomal changes, with five rearrangements occurring over approximately 7 million years, was recorded in the lineage of the small-toothed mole.

Sequencing of laser captured Z and W chromosomes of the tocantins paradoxical frog (Pseudis tocantins) provides insights on repeatome and chromosomal homology.

Pseudis tocantins is the only frog species of the hylid genus Pseudis that possesses highly heteromorphic sex chromosomes. Z and W chromosomes of Ps. tocantins differ in size, morphology, position of the nucleolar organizer region (NOR) and the amount and distribution of heterochromatin. A chromosomal inversion and heterochromatin amplification on the W chromosome were previously inferred to be involved in the evolution of this sex chromosome pair. Despite these findings, knowledge related to the molecular composition of the large heterochromatic band of this W chromosome is restricted to the PcP190 satellite DNA, and no data are available regarding the gene content of either the W or the Z chromosome of Ps. tocantins. Here, we sequenced microdissected Z and W chromosomes of this species to further resolve their molecular composition. Comparative genomic analysis suggests that Ps. tocantins sex chromosomes are likely homologous to chromosomes 4 and 10 of Xenopus tropicalis. Analyses of the repetitive DNA landscape in the Z and W assemblies allowed for the identification of several transposable elements and putative satellite DNA sequences. Finally, some transposable elements from the W assembly were found to be highly diverse and divergent from elements found elsewhere in the genome, suggesting a rapid amplification of these elements on the W chromosome.

Chromosome Comparisons of Australian Scaptodrosophila Species

Simple SummaryScaptodrosophila are a diverse group of flies that are widespread in Australia but have received limited research attention. In this study, we characterized the chromosomes of 12 Scaptodrosophila species. We found that the structural changes in their chromosomes are very similar to those seen in Drosophila, a related genus of flies. This includes the amplification of repetitive elements and changes in chromosome length in small (dot) chromosomes and the sex chromosomes. We also found numerous weak points along the arms of polytene chromosomes, which suggest the presence of internal repetitive sequences. Regions producing the nucleolus are at the same chromosome positions in Scaptodrosophila and Drosophila. These chromosomal studies provide a foundation for future genetic studies in Scaptodrosophila flies.The Scaptodrosophila represent a diverse group of Diptera closely related to Drosophila. Although they have radiated extensively in Australia, they have been the focus of few studies. Here, we characterized the karyotypes of 12 Scaptodrosophila species from several species groups and showed that they have undergone similar types of karyotypic change to those seen in Drosophila. This includes heterochromatin amplification involved in length changes of the sex and ‘dot’ chromosomes as well as the autosomes, particularly in the coracina group of species. Numerous weak points along the arms of the polytene chromosomes suggest the presence of internal repetitive sequence DNA, but these regions did not C-band in mitotic chromosomes, and their analysis will depend on DNA sequencing. The nucleolar organizing regions (NORs) are at the same chromosome positions in Scaptodrosophila as in Drosophila, and the various mechanisms responsible for changing arm configurations also appear to be the same. These chromosomal studies provide a complementary resource to other investigations of this group, with several species currently being sequenced.

The extensive amplification of heterochromatin in Melipona bees revealed by high throughput genomic and chromosomal analysis.

Satellite DNAs (satDNAs) and transposable elements (TEs) are among the main components of constitutive heterochromatin (c-heterochromatin) and are related to their functionality, dynamics, and evolution. A peculiar case regarding the quantity and distribution of c-heterochromatin is observed in the genus of bees, Melipona, with species having a low amount of heterochromatin and species with high amount occupying almost all chromosomes. By combining low-pass genome sequencing and chromosomal analysis, we characterized the satDNAs and TEs of Melipona quadrifasciata (low c-heterochromatin) and Melipona scutellaris (high low c-heterochromatin) to understand c-heterochromatin composition and evolution. We identified 15 satDNA families and 20 TEs for both species. Significant variations in the repeat landscapes were observed between the species. In M. quadrifasciata, the repetitive fraction corresponded to only 3.78% of the genome library studied, whereas in M. scutellaris, it represented 54.95%. Massive quantitative and qualitative changes contributed to the differential amplification of c-heterochromatin, mainly due to the amplification of exclusive repetitions in M. scutellaris, as the satDNA MscuSat01-195 and the TE LTR/Gypsy_1 that represent 38.20 and 14.4% of its genome, respectively. The amplification of these two repeats is evident at the chromosomal level, with observation of their occurrence on most c-heterochromatin. Moreover, we detected repeats shared between species, revealing that they experienced mainly quantitative variations and varied in the organization on chromosomes and evolutionary patterns. Together, our data allow the discussion of patterns of evolution of repetitive DNAs and c-heterochromatin that occurred in a short period of time, after separation of the Michmelia and Melipona subgenera.

Robertsonian rearrangements in Neotropical Meliponini karyotype evolution (Hymenoptera: Apidae: Meliponini).

Genome changes, evidenced through karyotype or nuclear genome size data, can result in reproductive isolation, diversification and speciation. The aim of this study was to understand how changes in the karyotype such as chromosome number and nuclear genome size accompanied the evolution of neotropical stingless bees, and to discuss these data in a phylogenetic context focusing on the karyotype evolution of this clade. We sampled 38 species representing the three Neotropical Meliponini groups; 35 for karyotype analyses and 16 for 1C value measurement. The chromosome number varied from 2n=16 to 2n=34, with distinct karyotypic formulae and the presence of a few polymorphisms, such as B chromosomes in one species and arm size differences between homologous chromosomes in two species. The mean 1C value varied from 0.31 pg to 0.92 pg. We associated empirical data on chromosome number and mean 1C value to highlight the importance of Robertsonian fusion rearrangements, leading to a decrease in chromosome number during the Neotropical Meliponini evolution. These data also allowed us to infer the independent heterochromatin amplification in several genera. Although less frequent, Melipona species with 2n=22 represent evidence of Robertsonian fissions. We also pointed out the importance of chromosomal rearrangements that did not alter chromosome number, such as inversions and heterochromatin amplification.

Different Levels of Chromatin Condensation in Partamona chapadicola and Partamona nhambiquara (Hymenoptera, Apidae)

Studies in several organisms have contributed to the understanding of heterochromatin and its biological importance. In bees of the tribe Meliponini, the presence of chromosomes with totally heterochromatic arms has been attributed to the mechanism of karyotype evolution in which this group accumulated heterochromatin to maintain telomere stability after centric fission events. In the present study, the use of classical and molecular cytogenetic techniques as well as automated image analysis software for the description of the karyotypes of Partamonachapadicola and P. nhambiquara bee species revealed variability in the compaction and patterns of chromatin structure. Although both species have the same chromosome number as other species in the genus Partamona (2n = 34), C-banding and image analyses indicated the existence of chromosomes with 3 regions of different staining intensities, suggesting a chromatin structure with distinct patterns and characteristics. Repetitive DNA probes hybridized only in the euchromatic regions, whereas the regions with intermediate staining intensity did not show any hybridization signals. This suggests that these regions present features more similar to heterochromatin. Evidence of the existence of a chromatin class with intermediate condensation compared to euchromatin and heterochromatin indicates a potential mechanism for heterochromatin amplification and demonstrates the need for further studies on this topic. This previously unrecognized class of chromatin should be taken into account in the study of all Meliponini chromosomes.

Insights on repetitive DNA behavior in two species of CtenusWalckenaer, 1805 and Guasuctenus Polotow and Brescovit, 2019 (Araneae, Ctenidae): Evolutionary profile of H3 histone, 18S rRNA genes and heterochromatin distribution.

Ctenidae represents one of the most representative spider families in the tropical forests of Brazil. Its largest genus, Ctenus, has approximately 220 species out of the more than 520 Ctenidae species described, and several authors consider it polyphyletic. Chromosomal data are only available for four species of Ctenus, representing a large gap in the cytogenetic knowledge about the group. This study provided cytogenetic data on two Ctenus species and one Guasuctenus (previously described as Ctenus). All showed 2n♂ = 28 (26+X1X20). Guasuctenus longipes presented two chromosome pairs containing 18S rDNA genes and C. medius, however C. ornatus showed only one chromosome pair with the 18S rDNA gene. Hybridization data using histone H3 probe indicated specific profiles: histone H3 genes were found in one chromosome pair in G. longipes, in three pairs in C. medius, and in four pairs in C. ornatus. Furthermore, supernumerary chromosomes were identified in C. ornatus presenting a meiotic behavior similar to that of sex chromosomes; and a trivalent was found in C. medius, formed by the association of one sex chromosome and an autosomal bivalent, indicating the importance of these events for the diversification of sex chromosomes in spiders. The C-banding pattern was similar between C. medius and C. ornatus with regard to the number and locations of heterochromatic bands, suggesting that heterochromatin amplification and dispersion, affect karyotypic evolution in the genus. Cytogenetic data showed similarity between C. medius and C. ornatus, and differentiation of G. longipes congruent with morphological data. Moreover, although more comparative analyses are needed to specify composition of the dispersed heterochromatin in Ctenus, the mapping of heterochromatic bands provided insights about the evolution of the karyotypes in this genus.

Repetitive DNAs in Melipona scutellaris (Hymenoptera: Apidae: Meliponidae): chromosomal distribution and test of multiple heterochromatin amplification in the genus

Melipona bees are remarkable due to the high contrast in heterochromatin amounts, making this group interesting for studying repetitive DNA amplification. Here, we performed the first efforts for the chromosomal localization of different repetitive DNAs in M. (Michmelia) scutellaris and tested for unique or multiple heterochromatin amplification in Melipona subgenera. Our data revealed enrichment of repetitive DNAs in chromosomal heterochromatic arms demonstrated by C0t-DNA and DOP-PCR probe hybridization, although microsatellites and multigene families were located at terminal euchromatic regions. Analysis using C0t-DNA probe from M. scutellaris showed positive hybridization only in Michmelia species, suggesting monophyletic amplification and sharing of heterochromatin sequences between species. However, the subgenus Melikerria, with a high amount of heterochromatin, probably underwent independent heterochromatin amplification or experienced sequence modification.

Genomic Organization of Repetitive DNAs and Differentiation of an XX/XY Sex Chromosome System in the Amazonian Puffer Fish, Colomesus asellus (Tetraodontiformes)

The genus Colomesus is the sole representative of the family Tetraodontidae in the Amazon region. Here, Colomesus asellus was analyzed using conventional and molecular cytogenetic protocols. Its diploid chromosome number is 2n = 46 with 12 meta-, 10 submeta-, 16 subtelo-, and 8 acrocentric chromosomes and a fundamental number of FN = 84. An XX/XY sex chromosome system was identified. Mapping of 18S rDNA correlated with the nucleolus organizer regions (Ag-NORs) in the short arms of the 2 X chromosomes in females and in the Y chromosome in males. C-banding revealed heterochromatin in the centromeric regions of all chromosomes, except for pair 3. Prominent sex chromosome-specific heterochromatin amplification was observed, covering the short arms of the Y chromosome almost entirely. FISH with telomeric and tropomyosin (tpm1) sequences, respectively, revealed terminal signals in all chromosomes. The analysis of extended DNA fibers confirmed the colocalization and the interspersed pattern of the telomeric and tpm1 sequences. Thus, this study highlights the remarkable evolutionary dynamism presented by the Amazonian puffer fish regarding the differentiation of a heteromorphic XY sex chromosome system and a particular sex-specific amplification of rDNA sites. This is the first record of such an association in the Tetraodontidae family.

Chromosomal diversity and molecular divergence among three undescribed species of Neacomys (Rodentia, Sigmodontinae) separated by Amazonian rivers.

The Neacomys genus (Rodentia, Sigmodontinae) is distributed in the Amazon region, with some species limited to a single endemic area, while others may occur more widely. The number of species within the genus and their geographical boundaries are not known accurately, due to their high genetic diversity and difficulties in taxonomic identification. In this work we collected Neacomys specimens from both banks of the Tapajós River in eastern Amazon, and studied them using chromosome painting with whole chromosome probes of Hylaeamys megacephalus (HME; Rodentia, Sigmodontinae), and molecular analysis using haplotypes of mitochondrial genes COI and Cytb. Chromosome painting shows that Neacomys sp. A (NSP-A, 2n = 58/FN = 68) and Neacomys sp. B (NSP-B, 2n = 54/FN = 66) differ by 11 fusion/fission events, one translocation, four pericentric inversions and four heterochromatin amplification events. Using haplotypes of the concatenated mitochondrial genes COI and Cyt b, Neacomys sp. (2n = 58/FN = 64 and 70) shows a mean divergence of 6.2% for Neacomys sp. A and 9.1% for Neacomys sp. B, while Neacomys sp. A and Neacomys sp. B presents a medium nucleotide divergence of 7.4%. Comparisons were made with other published Neacomys data. The Tapajós and Xingu Rivers act as geographic barriers that define the distribution of these Neacomys species. Furthermore, our HME probes reveal four synapomorphies for the Neacomys genus (associations HME 20/[13,22]/4, 6a/21, [9,10]/7b/[9,10] and 12/[16,17]) and demonstrate ancestral traits of the Oryzomyini tribe (HME 8a and 8b, 18 and 25) and Sigmodontinae subfamily (HME 15 and 24), which can be used as taxonomic markers for these groups.

Sex chromosome variability of Spermophilus citellus (Linnaeus, 1766) in the Southeastern part of the Balkan Peninsula

This work presents novel data that has allowed us to propose an evolutionary model for the karyotype of the European ground squirrel (Spermophilus citellus, L. 1766) based on sex chromosome variants. Routine C- and G-banding, specific fluorochrome staining (CMA3, DAPI) and FISH experiments were used to characterize the pattern of chromosomal variation in populations of S. citellus in the Southeastern part of the Balkan Peninsula, the ancestral area of the species with the highest genetic diversity. The established diploid chromosome number of S. citellus is 2n=40. All autosomes are biarmed: 12 pairs of large to small submetacentrics, 5 pairs of large to small subtelocentrics, one medium-sized and one small pair of metacentrics. Three different morphologies are observed for both X and Y chromosomes. The X chromosomes are classified as acrocentric, subtelocentric, or submetacentric. In most of the individuals studied, the Y chromosome was dot-like, with the exception of a few presenting small or medium-sized acrocentrics and metacentrics. Blocks of heterochromatin of different sizes were found in the centromeric regions of all autosomes and X chromosomes, while all Y chromosome variants appeared to be entirely heterochromatic. CMA3 fluorescence was bright in the majority of the C-positive regions and in the acrocentric X chromosome. These results were supported by DAPI staining, which showed that the CMA3-positive sites were DAPI-negative and approximately of the same distinct sizes. In situ hybridization with the (TTAGGG)n probe produced a staining typical of telomeric repeats at the edges of all chromosomes. Notably, the Y chromosome showed stronger accumulation of telomeric repeats than that observed on other chromosomes. The nature of the C-, G- and fluorochrome staining of the sex chromosomes of S. citellus suggested that the amplification of constitutive heterochromatin and the pericentric inversions were likely sufficient to explain the differences between the morphology of sex chromosome. The observed sex chromosome variability reinforces the differences among the populations of the S. citellus in the Southeastern part of the Balkan Peninsula and correlate with specific geographical areas. These will be further investigated in order to fully evaluate the adaptive value of chromosome rearrangements.

Evidence of separate karyotype evolutionary pathway in Euglossa orchid bees by cytogenetic analyses

Euglossini are solitary bees considered important pollinators of many orchid species. Information regarding chromosome organization is available for only a small number of species in this group. In the present work, the species Euglossa townsendi and E. carolina were analyzed by cytogenetic techniques to collect information that may aid the understanding of their evolution and chromosomal organization. The chromosome number found was n = 21 for males and 2n = 42 for females in the two species. The distribution and amount of heterochromatin regions differed in the two species analyzed, where they were classified as “high” or “low” heterochromatin content, similarly to what has already been performed in social bee species of the genus Melipona. Banding patterns found in this study suggest that other mechanisms may have occurred in the karyotype evolution of this group, unlike those suggested for social bees and ants. Karyotype evolution of solitary bees appears to have occurred as an event separate from other hymenopterans and did not involve chromosome fissions and heterochromatin amplification.

Chromosomal painting and ZW sex chromosomes differentiation in Characidium (Characiformes, Crenuchidae)

BackgroundThe Characidium (a Neotropical fish group) have a conserved diploid number (2n = 50), but show remarkable differences among species and populations in relation to sex chromosome systems and location of nucleolus organizer regions (NOR). In this study, we isolated a W-specific probe for the Characidium and characterized six Characidium species/populations using cytogenetic procedures. We analyzed the origin and differentiation of sex and NOR-bearing chromosomes by chromosome painting in populations of Characidium to reveal their evolution, phylogeny, and biogeography.ResultsA W-specific probe for efficient chromosome painting was isolated by microdissection and degenerate oligonucleotide primed-polymerase chain reaction (DOP-PCR) amplification of W chromosomes from C. gomesi. The W probe generated weak signals dispersed on the proto sex chromosomes in C. zebra, dispersed signals in both W and Z chromosomes in C. lauroi and, in C. gomesi populations revealed a proximal site on the long arms of the Z chromosome and the entire W chromosome. All populations showed small terminal W probe sites in some autosomes. The 18S rDNA revealed distinctive patterns for each analyzed species/population with regard to proto sex chromosome, sex chromosome pair, and autosome location.ConclusionsThe results from dual-color fluorescence in situ hybridization (dual-color FISH) using W and 18S rDNA probes allowed us to infer the putative evolutionary pathways for the differentiation of sex chromosomes and NORs, from structural rearrangements in a sex proto-chromosome, followed by gene erosion and heterochromatin amplification, morphological differentiation of the sex chromosomal pair, and NOR transposition, giving rise to the distinctive patterns observed among species/populations of Characidium. Biogeographic isolation and differentiation of sex chromosomes seem to have played a major role in the speciation process in this group of fish.

The HSR and heterochromatin distribution in natural populations ofMus musculus domesticus(Rodentia: Murinae) from Greece

Abstract The natural populations of the house mouse, Mus musculus domesticus, of the Euro-Mediterranean region have been under extensive scrutiny for several years, thanks to an extensive Robertsonian chromosomal variation. However, this subspecies is also known for the occasional appearance of Homogenously Staining Regions (HSRs) in chromosome 1 of the karyotype. Based on the extensive C-banding chromosomal study of 236 house mice from 32 localities of Peloponnisos, S Greece, eight Rb individuals from two localities were found with HSRs in their karyotype (HSR incidence: 3.4%), thus constituting the first record of this chromosomal trait in house mouse populations from Greece. Moreover in the context of this study, we came across five individuals with significant heterochromatin amplification below the centromere of the X chromosome and of two autosomal pairs (incidence: 2.1%). Notably, four of these individuals also carried HSRs. This finding is important because increase of heterochromatin content has ...

Heterogeneity of heterochromatin in six species ofCtenomys (Rodentia: Octodontoidea: Ctenomyidae) from Argentina revealed by a combined analysis of C- and RE-banding

Exceptional chromosomal variability makesCtenomys an excellent model for evolutionary cytogenetic analysis. Six species belonging to three evolutionary lineages were studied by means of restriction endonuclease and C-chromosome banding. The resulting banding patterns were used for comparative analysis of heterochromatin distribution on chromosomes. This combined analysis allowed intra- and inter-specific heterochromatin variability to be detected, groups of species belonging to different lineages to be characterized, and phylogenetic relationships hypothesized from other data to be supported. The “ancestral group”,Ctenomys pundti andC. talarum, share three types of heterochromatin, the most abundant of which was also found in C. aff.C. opimus, suggesting that the latter species also belongs to the “ancestral group”. Additionally, within the subspeciesC. t. talarum, putative chromosomal rearrangements distinguishing two of the three chromosomal races were identified. Two species belong to an “eastern lineage”,C. osvaldoreigi andC. rosendopascuali, and share only one type of heterochromatin homogeneously distributed across their karyotypes.C. latro, the only analyzed species from the “chacoan” lineage, showed three types of heterochromatin, one of them being that which characterizes the “eastern lineage”.C. aff.C. opimus, because of its low heterochromatin content, is the most primitive karyotype of the genus yet described. The heterochromatin variability showed by these species, reflecting the evolutionary divergence toward different heterochromatin types, may have diverged since the origin of the genus. Heterochromatin amplification is proposed as a trend withinCtenomys, occurring independently of chromosomal change in diploid numbers.

A repeat DNA sequence from the Y chromosome in species of the genus Microtus

In most mammals, the Y chromosome is composed of a large amount of constitutive heterochromatin. In some Microtus species, this feature is also extended to the X chromosome, resulting in enlarged (giant) sex chromosomes. Several repeated DNA sequences have been described in the gonosomal heterochromatin of these species, indicating that it has heterogeneous and species-specific composition and distribution. We have cloned an AT-rich, 851-bp long, repeated DNA sequence specific for M. cabrerae Y chromosome heterochromatin. The analysis of other species of the genus Microtus indicated that this sequence is also located on the Y chromosome (male-specific) in three species (M. agrestis, M. oeconomus and M. nivalis), present on both Y and X chromosomes and on some autosomes in M. arvalis and absent in the genome of M. guentheri. Our data also suggest that the mechanism of heterochromatin amplification operating on the sex chromosomes could have been different in each species since the repeated sequences of the gonosomal heterochromatic blocks in M. cabrerae and M. agrestis are different. The absence of this sequence in the mouse genome indicates that its evolutionary origin could be recent. Future analysis of the species distribution, localization and sequence of this repeat DNA family in arvicolid rodent species could help to establish the unsolved phylogenetic relationships in this rodent group.

Genetic variation and zoogeography of pine voles of the Microtus subterraneus/majori group in Europe and Asia Minor

AbstractKaryotypic and allozyme analyses were carried out on 55 individuals of Microtus subterraneus and M. majori from 19 populations in Europe and Anatolia. The occurrence of M. subterraneus in Asia Minor was proven. Standard karyotypes were found in M. majori (2n=54), M. subterraneus from Poland (2n=54), and central and south‐eastern Europe (2n=52). Although the karyotype of M. subterraneus populations from Anatolia (2n=54), resembled that from Poland, there were apparent differences in the X chromosomes: it was larger and submetacentric (instead of metacentric) in the Anatolian populations owing to the presence of a large block of heterochromatin. The heterochromatic segment was also found to cause the size and shape differences in the X chromosomes between M. majori and European M. subterraneus (both in the 2n=52 and 2n=54 karyotypes). However, the location of this segment differed between the latter species and M. subterraneus from Anatolia. The distribution of the 2n=52 and both the 2n=54 karyotypes of M. subterraneus suggests that the Robertsonian fusion, distinguishing these different karyotypes, originated within the range of the species (i.e. within the 54‐chromosome karyotype) and has spread subsequently towards its periphery. It is also suggested that both the fusion in European populations and amplification of heterochromatin in Anatolian populations are quite recent phenomena. The allozyme data indicate that gene flow is limited between populations of M. subterraneus and they are concordant with the isolation‐by‐distance model. There seems to be little correspondence between patterns of chromosomal and allozyme variation in M. subterraneus. Nei's genetic distance between M. subterraneus and M. majori was low (D=0.169), suggesting their recent divergence.

International study on Artemia. LXII. Genomic relationships between Artemia franciscana and A. persimilis, inferred from chromocentre numbers.

Chromocentres, i.e. heavily stainable heterochromatic areas with highly repetitive DNA (a 130-bp repeat in the order of 6 x 105 copies per haploid genome) observed in the resting nucleus, are a reliable taxonomic trait and a good marker for speciation in Artemia. This chromosome marker was evaluated in populations of two New World sibling species: A. franciscana, from North, Central and South America, and A. persimilis from Argentina. Artemia persimilis showed the characteristically low average chromocentre numbers (<3) of the ancestral Mediterranean species, whilst higher numbers (>5) were seen in A. franciscana. The increase in chromocentre numbers from A. persimilis to A. franciscana occurred in a rather steady, continuous geographical pattern with an overlapping zone between both species. A north-south steady latitudinal decline of chomocentres was observed towards the equator in A. franciscana populations from the northern hemisphere, whilst the trait increased from this point towards southern latitudes with a population in Chilean Patagonia (below 52 degrees S) exhibiting the highest chromocentre frequency. Since chromocentres are positively correlated with repetitive DNA content, differentiation between the species through the amplification of heterochromatin or an increased DNA content in A. franciscana (although this species has a lower diploid chromosome number), might nevertheless reflect chromosomal reorganizations between the species. Chromocentres, together with other traits, reveal a probable hybrid zone between the two species in Chilean Patagonia.