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Schubert, V.

Publications and source records attributed to Schubert, V..

At least 19 recordsLinked to original sources

Holocentromere diversity in Cyperaceae: contrasting repeat organisation in Mapanioideae and Cyperoideae

Centromeres ensure accurate chromosome segregation and are typically confined to a single, localised region in monocentric chromosomes. In contrast, holocentric chromosomes exhibit kinetochore activity distributed along the chromosome length. Although holocentricity is widespread in Cyperaceae, the composition and organisation of these centromeres, as well as their evolutionary diversification, remain poorly understood. Here, we investigated centromere organisation in representatives of the subfamilies Mapanioideae (Hypolytrum schraderianum Nees) and Cyperoideae (Cladium mariscus (L.) Pohl) by combining genome assemblies, repeatome characterisation (RepeatExplorer), fluorescence in situ hybridisation (FISH), and immunolocalisation. Comparative synteny analyses incorporating the genomes of Rhynchospora breviuscula (n = 5) and Carex littledalei (n = 29) identified conserved blocks, eventually expanding almost whole chromosomes of H. schraderianum (n = 30) and Cl. mariscus (n = 39), despite divergent chromosome numbers and deep evolutionary distances within Cyperaceae. Mobile elements showed very low abundances and were uniformly dispersed, with Ty1/Copia Angela being the most abundant in both species. In Cl. mariscus, holocentromeres showed an extended distribution of centromere- and kinetochore-associated proteins along the chromosomes, largely colocalised with two satellite DNA repeats that form dispersed clusters. In contrast, H. schraderianum also displayed kinetochore signals along chromatids, but the most abundant satellite DNA family was enriched in distal and interstitial chromosomal regions rather than interspersed along the chromatids. Together, these results reveal different genomic architectures underlying holocentric organisation in phylogenetically distinct Cyperaceae lineages, suggesting that holocentromeres in this family have diversified with variation in centromere organisation in regard to its association with repetitive DNA.

evolutionary biology↗

Nascent transcripts of LL2R tandem repeat nucleate locus-specific RNP condensates recruiting splicing factors

Spatial organization of the cell nucleus is increasingly understood to be governed by non-coding architectural RNAs that seed the formation of biomolecular condensates. However, the genomic origins and mechanistic principles by which specific architectural RNAs orchestrate ribonucleoprotein (RNP) compartments remain poorly defined. Here, we characterised a novel non-coding architectural RNA that nucleates the formation of a locus-associated membraneless nuclear compartment. Using a combination of T2T genomic, transcriptomic, and high-resolution mapping on giant lampbrush chromosomes, we identified an array of non-coding LL2R tandem repeats on the long arm of chicken chromosome 2. The LL2R tandem repeat array is slowly transcribed by a convoy of RNA polymerases II as a single transcription unit from a retrotransposon promoter. With the use of scanning electron and super-resolution microscopy for detailed analysis, we demonstrate that nascent LL2R repeat transcripts are retained at the site of their transcription and recruit multiple sets of core splicing factors, including TMG-capped snRNAs, core snRNP proteins, and serine/arginine-rich proteins SRRM2 and SC35 that contain intrinsically disordered regions, thus seeding nuclear domain formation. At the same time, the RNP-matrix of the LL2R repeat transcription unit lacks hnRNP L, hnRNP K, and SFPQ, widely distributed along normal transcription loops that bear nascent gene transcripts. In contrast to pre-mRNAs, strand-specific LL2R tandem repeat-containing RNA contains more than threefold excess of predicted donor splice sites compared to acceptor splice sites and is depleted in polyadenylation signals. We suggest that stalling of elongating RNA polymerases at LL2R transcription units and delay in 3 end cleavage may be caused by a deficiency of co-transcriptional splicing of nascent LL2R repeat transcripts. LL2R repeat-containing RNA also forms non-diffusible RNP-aggregates enriched by splicing factors that could serve as a site of post-transcriptional splicing. Our findings reveal how tandem repeat-derived RNAs drive the assembly of oocyte nuclear condensates, providing a functional template for understanding the formation of closely related nuclear speckles and nuclear stress bodies across eukaryotes.

cell biology↗

Newly unveiled meiosis elucidates the unreduced gamete frequency and its impact on evolution of the Lemna minor complex

Fusion of gametes possessing meiotically reduced (haploid) chromosome complements is the main pathway of propagation among eukaryotes. However, duckweeds, the smallest angiosperms, propagate mainly vegetatively, and meiosis has not yet been documented in detail for this plant family. The more surprising was the recent evidence of rather frequent interspecific hybrids and triploid clonal accessions which became obvious by genome size measurements, genomic in situ hybridization (GISH) and combined plastid and nuclear DNA markers. These observations indicated sexual propagation involving reduced as well as unreduced male and female gametes in Lemna minor and L. turionifera leading to allodiploid and allotriploid hybrids (MT, MMT, MTT) and autotriploid L. minor (MMM) accessions. Here, we i) documented the meiotic stages of Lemna species for the first time; ii) provided evidence of unreduced male gametes through fluorescent in situ hybridization (FISH) with single locus probes; iii) determined their abundance in different individuals and iv) hypothesized about the reasons of unreduced male gamete formation. These findings open new insights into the modes of sexual reproduction and evolution of duckweeds which may be useful for future breeding efforts in this emerging crop.

plant biology↗

The C-terminal SUMOylation-dependent regulation of αKNL2 governs its centromere targeting and interaction with CENH3

The centromere is a specialized domain that facilitates chromosome segregation during mitosis and serves as the site for kinetochore formation. KINETOCHORE NULL2 (KNL2) is essential for the recognition and loading of the centromeric histone H3 variant, CENH3, to centromeres. A yeast two-hybrid screen for KNL2 interactors identified components of the SUMOylation pathway. However, the role of KNL2 SUMOylation in Arabidopsis has not yet been determined. In this study, we demonstrated that the C-terminal part of KNL2 interacts with SUMO3 and ULP1d, as shown by BiFC and co-immunoprecipitation assays. Bioinformatic and functional analysis identified three SUMOylation and two SUMO-interacting motif (SIM) sites in the C-terminal region of KNL2, which are critical for growth, fertility, and chromosome alignment. Of the three SUMOylation sites, Lys474 and Lys511 were the most critical for the centromeric localization of KNL2, underscoring the importance of KNL2 SUMOylation for its function. Additionally, both in vitro and in vivo assays showed that KNL2-C undergoes SUMOylation by SUMO1 or SUMO3. The SUMO protease mutant, ulp1d-2 led to the slight accumulation of SUMOylated KNL2 in Arabidopsis. We further showed that SUMOylation of KNL2 promotes its binding to CENH3 and controls protein stability. Our findings show that C-terminal SUMOylation of KNL2 is crucial for its centromeric localization, interaction with CENH3, and kinetochore assembly, emphasizing the significance of post-translational modifications in chromosome segregation and cell division in plants.

molecular biology↗

Hybridity of mainly asexually propagating duckweeds in genus Lemna - dead end or breakthrough?

O_LIThe cosmopolitan, mainly vegetatively propagating, organ-reduced monocotyledonous aquatic duckweeds are the smallest and fastest growing angiosperms, distributed world-wide and flower rarely in nature. Recently, we reported intra- and interspecific hybrids and ploidy variants in the genus Lemna. Thus, contrary to the expectation, sexual propagation may occasionally occur within and between Lemna species. Our main goal was to uncover whether the ecologically successful hybrids are evolutionary dead ends, or initiate further speciation and novel sexual recombination. C_LIO_LIWe investigated flower development, pollen viability, seed set and seed germination in hybrids and their parental species and characterized genome size and genetic markers in the progenies. C_LIO_LIIntraspecific crosses yielded fertile progeny, but all dihaploid and triploid interspecific hybrids were male sterile. Only an established allotetraploid hybrid reproduced sexually, while colchicine-induced allotetraploids from dihaploids did not re-gain sexual competence so far. C_LIO_LIWe concluded that only established allotetraploid hybrids represent an evolutionary break-through in duckweeds. Our results regarding sexual traits within the duckweed genus Lemna, and the sexual competence of diverse hybrids i) pave the way for further investigation in this understudied field, ii) provide fundamental data regarding the evolutionary potential of duckweed hybrids and iii) are important for future breeding efforts on this emerging crop. C_LI

evolutionary biology↗

Kinetochore mutations and histone phosphorylation pattern changes preceded holo- and macro-monocentromere evolution

Centromeres are essential for kinetochore assembly and spindle attachment. While chromosomes of most species are monocentric with a single centromere, a minority exhibit holocentricity, with a centromere along the chromatid length. Sporadic emergence of holocentricity suggests multiple independent transitions. To explore this, we compared the centromere and (epi)genome organization of two sister genera with contrasting centromere types: Chamaelirium luteum with large "macro-monocentromeres" and Chionographis japonica with holocentromeres. Both exhibit chromosome-wide histone phosphorylation patterns distinct from typical monocentric species. Kinetochore analysis revealed similar chimeric Borealin in both species, with additional KNL2 loss and NSL1 chimerism in Cha. luteum. The broad-scale synteny between both genomes supports de novo holocentromere formation in Chi. japonica. Despite sharing features with both centromere types, macro-monocentromeres do not represent a direct link between mono- and holocentromeres. We propose a model for the divergent evolution involving kinetochore gene mutations, altered histone phosphorylation patterns, and centromeric satellite DNA amplification.

evolutionary biology↗

Triploidy is prominent in the duckweed Lemna minor complex

Duckweeds (Lemnaceae Martinov) are aquatic monocotyledonous flowering plants comprising five genera and 35 recognized species, known for being the smallest and fastest-growing flowering plants on Earth. Many species are morphologically indistinguishable due to their highly reduced structures, yet molecular evidence suggests that visually similar clones may represent distinct species or hybrids. For example, clonal accessions of the globally distributed Lemna minor in the Landolt Duckweed Collection exhibit genome size variations of several hundred megabases (Mb), raising questions about their taxonomic classification and evolutionary origins. We analyzed 58 presumed L. minor clones to resolve these relationships using a comprehensive suite of methods, including whole-genome sequencing (WGS), flow-cytometric genome size measurements, molecular markers, chromosome counting, and genomic in situ hybridization (GISH). Our findings reveal extensive genome plasticity within the "Lemna minor complex," identifying diploid and triploid L. minor clones, as well as di-haploid and triploid interspecific hybrids called L. x japonica (L. minor x L. turionifera), L. x mediterranea (L. minor x L. gibba), and a novel African-clade distinct from known L. minor lineages. Triploidy was prevalent, occurring in 29% of the clones, and was associated with enhanced growth under optimal conditions but reduced performance under high light and temperature. These findings highlight the widespread role of triploidy, cryptic species, and hybridization in the L. minor complex, emphasizing the importance of multiple approaches for accurately classifying duckweed species and understanding their evolutionary trajectories.

plant biology↗

Complete genome sequence assembly elucidates evolution and regulation of 5S rDNA loci in the greater duckweed Spirodela polyrhiza

We resolved the molecular architecture of the 5S rDNA loci in an aquatic monocot Spirodela polyrhiza. Measurements of fluorescence in situ hybridization signals revealed two loci with 5S rDNA clusters. A combination of the extra-long DNA reads and conventional sequencing of 5S rDNA repeats allowed the assembling of complete loci sequences located on ChrSp6 and ChrSp13. The homologous chromosomes of the ChrSp6 locus contain clusters of 40 and >60 copies of 5S rDNA repeat units with intergenic spacer (IGS) of 400 bp. The ChrSp13 locus is represented by 5S rDNA clusters of rRNA genes spaced by IGSs of 1,056 or 1,069 bp arranged in two sub-clusters, suggesting a different rate of repeat homogenization between loci. The G/C-rich 5S rDNA arrays in both loci are embedded in A/T-enriched chromosome regions with possible regulatory functions. The TATA-like boxes of the 5S rDNA repeat on ChrSp6 and ChrSp13 exhibit different affinities for the TATA-binding protein in 3D modeling of protein/DNA interactions, suggesting a locus-specific regulation of rRNA transcription. Our findings shed light on the molecular architecture of the 5S rDNA loci, which could foster an innovative era for the principles of evolution and regulation of rRNA genes in plants.

molecular biology↗

Genome diversity and phylogeny of the section Alatae of genus Lemna (Lemnaceae), comprising the presumed species Lemna aequinoctialis, Le. perpusilla and Le. aoukikusa

O_LIThe Section Alatae of genus Lemna of the monocotyledonous aquatic duckweed family (Lemnaceae) consists of rather diverse accessions with unknown phylogeny and unclear taxonomic assignment. In contrast to other duckweeds, some Alatae accessions, in addition to mainly vegetative propagation, produce readily flowers and viable seeds. We analyzed the genomic diversity and phylogenetic relationship of 52 Alatae accessions. C_LIO_LIFor this purpose, we applied multiple molecular and cytogenetic approaches, including plastid and nuclear sequence polymorphisms, chromosome counting, genome size determination and genomic in situ hybridization in combination with geographic distribution. C_LIO_LIWe uncovered ploidy variation, recurrent hybridization, and backcrosses between diploid species and their hybrids. The latter successfully spread over three continents. C_LIO_LIThe results elucidate the evolution of Alatae accessions (summarized in Fig. 7) and explain the difficult taxonomic assignment of distinct accessions. Our study might be an example for analogous studies to resolve the hitherto unclear relationships among accessions of the duckweed genera Wolffiella and Wolffia. C_LI O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=166 SRC="FIGDIR/small/632899v1_fig7.gif" ALT="Figure 7"> View larger version (23K): org.highwire.dtl.DTLVardef@d05f08org.highwire.dtl.DTLVardef@f33c5org.highwire.dtl.DTLVardef@ff7adorg.highwire.dtl.DTLVardef@4da6e4_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOFigure 7C_FLOATNO Hypothetical scenario of evolution within Lemna Section Alatae. In bold are accessions from America. The classically defined diploid species Le. aequinoctialis and Le. perpusilla sensu stricto were confirmed by the applied molecular methods. The previously named Le. aoukikusa accessions were uncovered as tetraploid interspecific hybrids between Le. aequinoctialis and Le. perpusilla. In addition, further tri- and tetraploid hybrids as well as tri- and tetraploid "Backcross hybrids" were detected from the USA and China, respectively. C_FIG

evolutionary biology↗

Functional monocentricity with holocentric characteristics andchromosome-specific centromeres in a stick insect

Centromeres are specialized genomic regions that are critical for chromosome segregation in eukaryotes. However, how the diversity of centromeres evolved, ranging from localized centromeres in monocentric species to complex chromosome-wide distributed centromere regions in holocentric species, remains elusive. Our cellular and genomic analyses in Timema stick insects reveal that within cell variation of the major centromere protein CenH3 recapitulates the variation typically observed across species. While CenH3-containing nucleosomes are distributed in a monocentric fashion on autosomes and bind tandem repeat sequences specific to individual or small groups of chromosomes, they exhibit a holocentric-like distribution on the sex chromosome and bind to more complex intergenic regions. Despite this contrasting distribution, all chromosomes, including the sex chromosome, attach to spindle microtubules at a single location, marking the first instance of a functionally monocentric species with holocentric-like attributes. Together, our findings highlight the potential for gradual transitions towards holocentricity or CenH3-independent centromere determination, and help to understand the rapid centromere sequence divergence between species.

evolutionary biology↗

Repeat-based holocentromeres of the woodrush Luzula sylvatica reveal new insights into the evolutionary transition from mono- to holocentricity

Although the centromere is restricted to a single region of the chromosome in most studied eukaryotes, members of the rush family (Juncaceae) harbor either monocentric (Juncus) or holocentric (Luzula) chromosomes. This provides an opportunity to study the evolutionary mechanisms involved in the transition to holocentricity. Here by combining chromosome-scale genome assembly, epigenetic analyses, immuno-FISH, and super-resolution microscopy, we report the occurrence of repeat-based holocentromeres in L. sylvatica. We found an irregular distribution of genes, centromeric units, and most repeats along the chromosomes. We determined the centromere function predominantly associated with two satellite DNA repeats, Lusy1 and Lusy2 of 124- and 174-bp monomer length, respectively, while CENH3 also binds satellite-free gene-poor regions. Comparative repeat analysis revealed that Lusy1 is present in most Luzula species, suggesting a conserved centromere role of this repeat. Synteny between L. sylvatica (n = 6) and J. effusus (n = 21) genomes further evidenced a chromosome number reduction in Luzula derived from multiple chromosome fusions of ancestral J. effusus-like chromosomes. We propose that the transition to holocentricity in Luzula involves: (i) fusion of small chromosomes resembling Juncus-like centromeres; (ii) expansion of atypical centromeric units; and (iii) colonization of satellite DNA for centromere stabilization.

genomics↗

Meiotic DSB repair DNA synthesis tracts in Arabidopsis thaliana

We report here the successful labelling of meiotic prophase I DNA synthesis in the flowering plant, Arabidopsis thaliana. Incorporation of the thymidine analogue, EdU, enables visualisation of the footprints of recombinational repair of programmed meiotic DNA double-strand breaks (DSB), with [~]400 discrete, SPO11-dependent, EdU-labelled chromosomal foci clearly visible at pachytene and later stages of meiosis. This number equates well with previous estimations of 200-300 DNA double-strand breaks per meiosis in Arabidopsis, confirming the power of this approach to detect the repair of most or all SPO11-dependent meiotic DSB repair recombination. The chromosomal distribution of these DNA-synthesis foci accords with that of early recombination markers and MLH1, which marks Class I crossover sites, colocalises with the EdU foci. It is currently estimated that [~]10 cross-overs (CO) and an equivalent number of non-cross-overs (NCO) occur in each Arabidopsis male meiosis. Thus, at least 90% of meiotic recombination events, and very probably more, have not previously been accessible for analysis. Visual examination of the patterns of the foci on the synapsed pachytene chromosomes corresponds well with expectations from the different mechanisms of meiotic recombination and notably, no evidence for long Break-Induced Replication DNA synthesis tracts was found. Labelling of meiotic prophase I, SPO11-dependent DNA synthesis holds great promise for further understanding of the molecular mechanisms of meiotic recombination, at the heart of reproduction and evolution of eukaryotes. Author SummarySexual reproduction involves the fusion of two cells, one from each parent. To maintain a stable chromosome complement across generations, these specialized reproductive cells must be produced through a specialized cell division called meiosis. Meiosis halves the chromosome complement of gametes and recombines the parental genetic contributions in each gamete, generating the genetic variation that drives evolution. The complex mechanisms of meiotic recombination have been intensely studied for many years and we now know that it involves the repair of programmed chromosomal breaks through recombination with intact template DNA sequences on another chromatid. At the molecular level, this is known to involve new DNA synthesis at the sites of repair/recombination and we report here the successful identification and characterisation of this DNA neo-synthesis during meiosis in the flowering plant, Arabidopsis. Both the characteristics and numbers of these DNA synthesis tracts accord with expectations from theory and earlier studies. Potentially applicable to studies in many organisms, this approach provides indelible footprints in the chromosomes and has the great advantage of freeing researchers from dependence on indirect methods involving detection of proteins involved in these dynamic processes.

genetics↗

KNL1 and NDC80 represent new universal markers for the detection of functional centromeres in plants

Centromere is the chromosomal site of kinetochore assembly and microtubule attachment for chromosome segregation. Given its importance, markers that allow specific labeling of centromeric chromatin throughout the cell cycle and across all chromosome types are sought for facilitating various centromere studies. Antibodies against the N-terminal region of CENH3 are commonly used for this purpose, since CENH3 is the near-universal marker of functional centromeres. However, because the N-terminal region of CENH3 is highly variable among plant species, antibodies directed against this region usually function only in a small group of closely related species. As a more versatile alternative, we present here antibodies targeted to the conserved domains of two outer kinetochore proteins, KNL1 and NDC80. Sequence comparison of these domains across more than 350 plant species revealed a high degree of conservation, particularly within a six amino acid motif, FFGPVS in KNL1, suggesting that both antibodies would function in a wide range of plant species. This assumption was confirmed by immunolabeling experiments in angiosperm (monocot and dicot) and gymnosperm species, including those with mono-, holo-, and meta-polycentric chromosomes. In addition to centromere labeling on condensed chromosomes during cell division, both antibodies detected the corresponding regions in the interphase nuclei of most species tested. These results demonstrated that KNL1 and NDC80 are better suited for immunolabeling centromeres than CENH3, because antibodies against these proteins offer incomparably greater versatility across different plant species which is particularly convenient for studying the organization and function of the centromere in non-model species.

genetics↗

Global nuclear reorganization during heterochromatin replication in the giant-genome plant Nigella damascena L.

Among flowering plants, genome size varies remarkably, by >2200-fold, and this variation depends on the loss and gain of non-coding DNA sequences that form distinct heterochromatin complexes during interphase. In plants with giant genomes, most chromatin remains condensed during interphase, forming a dense network of heterochromatin threads called interphase chromonemata. Using super-resolution light and electron microscopy, we studied the ultrastructure of chromonemata during and after replication in root meristem nuclei of Nigella damascena L. During S-phase, heterochromatin undergoes transient decondensation locally at DNA replication sites. Due to the abundance of heterochromatin, the replication leads to a robust disassembly of the chromonema meshwork and a general reorganization of the nuclear morphology visible even by conventional light microscopy. After replication, heterochromatin recondenses, restoring the chromonema structure. Thus, we show that heterochromatin replication in interphase nuclei of giant-genome plants induces a global nuclear reorganization.

cell biology↗

The genomes and epigenomes of aquatic plants (Lemnaceae) promote triploid hybridization and clonal reproduction

The Lemnaceae (duckweeds) are the worlds smallest but fastest growing flowering plants. Prolific clonal propagation facilitates continuous micro-cropping for plant-based protein and starch production, and holds tremendous promise for sequestration of atmospheric CO2. Here, we present chromosomal assemblies, annotations, and phylogenomic analysis of Lemna genomes that uncover candidate genes responsible for the metabolic and developmental traits of the family, such as anatomical reduction, adaxial stomata, lack of stomatal closure, and carbon sequestration via crystalline calcium oxalate. Lemnaceae have selectively lost genes required for RNA interference, including Argonaute genes required for reproductive isolation (the triploid block) and haploid gamete formation. Triploid hybrids arise commonly among Lemna, and we have found mutations in highly-conserved meiotic crossover genes that could support polyploid meiosis. Syntenic comparisons with Wolffia and Spirodela reveal that diversification of these genera coincided with the "Azolla event" in the mid-Eocene, during which aquatic macrophytes reduced high atmospheric CO2 levels to those of the current ice age. Facile regeneration of transgenic fronds from tissue culture, aided by reduced epigenetic silencing, makes Lemna a powerful biotechnological platform, as exemplified by recent engineering of high-oil Lemna that outperforms oil seed crops.

plant biology↗

Disruption of the standard kinetochore in holocentric Cuscuta species

Segregation of chromosomes depends on the centromere. Most species are monocentric, with the centromere restricted to a single region per chromosome. In some organisms, monocentric organization changed to holocentric, in which the centromere activity is distributed over the entire chromosome length. However, the causes and consequences of this transition are poorly understood. Here, we show that the transition in the genus Cuscuta was associated with dramatic changes in the kinetochore, a protein complex that mediates the attachment of chromosomes to microtubules. We found that in holocentric Cuscuta species the KNL2 genes were lost; the CENP-C, KNL1, and ZWINT1 genes were truncated; the centromeric localization of CENH3, CENP-C, KNL1, MIS12, and NDC80 proteins was disrupted; and the spindle assembly checkpoint (SAC) was degenerated. Our results demonstrate that holocentric Cuscuta species lost the ability to form a standard kinetochore and do not employ SAC to control the attachment of microtubules to chromosomes.

genetics↗

Plasticity in centromere organization: Holocentromeres can consist of merely a few megabase-sized satellite arrays

The centromere is the chromosome region where the microtubules attach during cell division. In contrast to monocentric chromosomes with one centromere location, holocentric species usually distribute hundreds of centromere units along the entire chromatid. We assembled the chromosome-scale reference genome and analyzed the holocentromere and (epi)genome organization of the lilioid Chionographis japonica. Remarkably, each of its holocentric chromatids consists of only 7 to 11 evenly-spaced megabase-sized centromere-specific histone H3-positive units. These units contain satellite arrays of 23 and 28 bp-long monomers capable of forming palindromic structures. Like monocentric species, C. japonica forms distinctly clustered centromeres in chromocenters at interphase. Additionally, the large-scale eu- and heterochromatin arrangement differs between C. japonica and other known holocentric species. Using polymer simulations, we modeled the formation of prometaphase line-like holocentromeres from interphase centromere clusters. Our findings broaden the knowledge about the diversity of centromere organization, showing that holocentricity is not restricted to species with numerous and small centromere units.

plant biology↗

Helical metaphase chromatid coiling is conserved

The higher-order organization of metaphase chromosomes has been debated for almost 140 years. Classical light and electron microscopy studies suggested that chromatids are composed of helically organized chromatin fibers (chromonemata). Non-helical models were also recently proposed. We studied chromosome organization in barley using cutting-edge approaches and obtained evidence for a helically arranged 400-nm chromatin fiber representing the chromonema within chromatid arms. The number of turns is positively correlated with arm length. Turn size and chromatin density decrease towards the telomeres. Due to their specialized functions, the helical organization of centromeres and nucleolus-organizing regions is interrupted by several thinner, straight chromatin fibers. A comparison with previously published data indicates that the helical turning of metaphase chromatid arms is a conserved feature of large eukaryotic chromosomes.

cell biology↗