Flow sorting of C-genome chromosomes from wild relatives of wheatAegilops markgrafii,Ae. triuncialisandAe. cylindrica, and their molecular organization

Molnár, István; Vrána, Jan; Farkas, András; Kubaláková, Marie; Cseh, András; Molnár-Láng, Márta; Doležel, Jaroslav

doi:10.1093/aob/mcv073

Cited by 34 publications

(24 citation statements)

References 73 publications

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“…speltoides and Ae. markgrafii were analyzed for the distribution of DAPI fluorescence intensity (flow karyotypes), narrow peaks were obtained, giving better chromosome resolution as compared to our previous work (Molnár et al ., , , ). This was probably due to the fact that we used a BD FACSAria II SORP flow sorter in this study, which employs a gel‐coupled flow cell instead of the classic jet‐in‐air system of BD FACSVantage flow sorter.…”

Section: Resultsmentioning

confidence: 99%

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Dissecting the U, M, S and C genomes of wild relatives of bread wheat (Aegilops spp.) into chromosomes and exploring their synteny with wheat

et al. 2016

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SIGNIFICANCE STATEMENT (74WORDS)Bivariate flow cytometric analysis of DNA content and FITC-labelled microsatellites enabled all the chromosomes in the U, M, S and C genomes ofAegilopsto be discriminated and purified. Mapping COS markers with known position in the wheat genome to flow-sorted Aegilops chromosomes revealed significant evolutionary rearrangements in the U and C genomes, but not in the M and S genomes. COS markers assigned to Aegilops chromosomes will facilitate alien introgression breeding in wheat.

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Section: Resultsmentioning

confidence: 99%

“…speltoides (Molnár et al ., ), chromosome 4C from Ae. markgrafii (Molnár et al ., ) and chromosomes 1U, 3U and 6U from Ae. umbelulata (Molnár et al ., ).…”

Section: Discussionmentioning

confidence: 99%

Dissecting the U, M, S and C genomes of wild relatives of bread wheat (Aegilops spp.) into chromosomes and exploring their synteny with wheat

et al. 2016

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“…The role of presynaptic centromere associations and their transition to homologous pairing Individual chromosome pairs visualized by in situ hybridization (Schwarzacher et al, 1992) can be studied in alien introgression lines of hexaploid wheat, carrying whole chromosomes or chromosome arms from a related species (Moln ar et al, 2015;Ali et al, 2016). A study on a wheat-rye translocation line revealed that cognition of homologous chromosome partners initiates at the centromeres in meiotic interphase and, subsequently, homologous chromosome segments align in leptotene with synapsis starting at the telomeres (Schwarzacher, 1997).…”

Section: Discussionmentioning

confidence: 99%

CENH3 morphogenesis reveals dynamic centromere associations during synaptonemal complex formation and the progression through male meiosis in hexaploid wheat

et al. 2017

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SUMMARYDuring meiosis, centromeres in some species undergo a series of associations, but the processes and progression to homologous pairing is still a matter of debate. Here, we aimed to correlate meiotic centromere dynamics and early telomere behaviour to the progression of synapotonemal complex (SC) construction in hexaploid wheat (2n=42) by triple immunolabelling of CENH3 protein marking functional centromeres, and SC proteins ASY1 (unpaired lateral elements) and ZYP1 (central elements in synapsed chromosomes). We show that single or multiple centromere associations formed in meiotic interphase undergo a progressive polarisation (clustering) at the nuclear periphery in early leptotene, leading to formation of the telomere bouquet. Critically, immunolabelling shows the dynamics of these presynaptic centromere associations and a structural reorganisation of the centromeric chromatin coinciding with key events of synapsis initiation from the subtelomeric regions. As short stretches of subtelomeric synapsis emerged at early zygotene, centromere clusters lost their strong polarization, gradually resolving as individual centromeres indicated by more than 21 CENH3 foci associated with unpaired lateral elements. Only following this centromere depolarisation were homologous chromosome arms connected, as observed by the alignment and fusion of interstitial ZYP1 loci elongating at zygotene so synapsis at centromeres is a continuation of the interstitial synapsis. Our results thus reveal that centromere associations are a component of the timing and progression of chromosome synapsis, and the gradual release of the individual centromeres from the clusters correlates with the elongation of interstitial synapsis between the corresponding homologues.

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“…DNA preparations from flow-sorted chromosomes can then be used for the generation of chromosome-specific sequences and assemblies [45, 46]. Application of the chromosome banding technique presented here should provide unambiguous identification of flow sorted chromosomes and help to reveal contamination.…”

Section: Discussionmentioning

confidence: 99%

Fluorescence chromosome banding and FISH mapping in perennial ryegrass, Lolium perenne L.

et al. 2016

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BackgroundThe unambiguous identification of individual chromosomes is a key part of the genomic characterization of any species. In this respect, the development and application of chromosome banding techniques has revolutionised mammalian and especially, human genomics. However, partly because of the traditional use of chromosome squash preparations, consistent fluorescence banding has rarely been achieved in plants. Here, successful fluorescence chromosome banding has been achieved for the first time in perennial ryegrass (Lolium perenne), a forage and turf grass with a large genome and a symmetrical karyotype with chromosomes that are difficult to distinguish.ResultsBased on flame-dried chromosome preparations instead of squashes, a simple fluorescence Q-banding technique using quinacrine mustard, unambiguously identified each chromosome and enabled the development of a banded karyotype and ideogram of the species. This Q-banding technique was also shown to be compatible with sequential FISH mapping enabling labelled genes and molecular markers to be precisely assigned to specific cytogenetic bands. A technique for DAPI-banding, which gave a similar pattern to Q-banding, was also introduced. This was compatible with FISH mapping and was used to anchor a single copy gene from an earlier mapped linkage group of L. perenne, thus providing a step towards integration of the genetic and cytogenetic maps.ConclusionsBy enabling the allocation of genes mapped by other methods to physically identified chromosome positions, this work will contribute to a better understanding of genomic structures and functions in grasses.

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Flow sorting of C-genome chromosomes from wild relatives of wheatAegilops markgrafii,Ae. triuncialisandAe. cylindrica, and their molecular organization

Cited by 34 publications

References 73 publications

Dissecting the U, M, S and C genomes of wild relatives of bread wheat (Aegilops spp.) into chromosomes and exploring their synteny with wheat

Dissecting the U, M, S and C genomes of wild relatives of bread wheat (Aegilops spp.) into chromosomes and exploring their synteny with wheat

CENH3 morphogenesis reveals dynamic centromere associations during synaptonemal complex formation and the progression through male meiosis in hexaploid wheat

Fluorescence chromosome banding and FISH mapping in perennial ryegrass, Lolium perenne L.

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