ArticlePloS one2013
FISHIS: fluorescence in situ hybridization in suspension and chromosome flow sorting made easy.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 51 papers.
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51 citing papers in PubMed, 99 citations in OpenAlex.
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- Dissection of a rapidly evolving wheat resistance gene cluster by long-read genome sequencing accelerated the cloning of Pm69.Plant communications · 2024Article
- The wheat stem rust resistance gene Sr43 encodes an unusual protein kinase.Nature genetics · 2023Article
- Flow Cytometric Analysis and Sorting of Plant Chromosomes.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Critical Steps in DAPI and FISH Imaging of Chromosome Spread Preparations.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Isolation and Sequencing of Chromosome Arm 7RS of Rye,International journal of molecular sciences · 2022Article
- Isolation and sequencing of a single copy of an introgressed chromosome from a complex genome for gene and SNP identification.TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik · 2022Article
- Aegilops sharonensis genome-assisted identification of stem rust resistance gene Sr62.Nature communications · 2022Article
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- Identification, High-Density Mapping, and Characterization of New Major Powdery Mildew Resistance Loci From the Emmer Wheat Landrace GZ1.Frontiers in plant science · 2022Article
- In-Depth Sequence Analysis of Bread WheatInternational journal of molecular sciences · 2021Article
- Chromosome analysis and sorting.Cytometry. Part A : the journal of the International Society for Analytical Cytology · 2021Review
- A membrane-bound ankyrin repeat protein confers race-specific leaf rust disease resistance in wheat.Nature communications · 2021Article
- Fundamentally different repetitive element composition of sex chromosomes in Rumex acetosa.Annals of botany · 2021Article
- Development of DNA Markers From Physically Mapped Loci inFrontiers in plant science · 2021Article
- A Cross between Bread Wheat and a 2D(2R) Disomic Substitution Triticale Line Leads to the Formation of a Novel Disomic Addition Line and Provides Information of the Role of Rye Secalins on Breadmaking Characteristics.International journal of molecular sciences · 2020Article
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The large size and complex polyploid nature of many genomes has often hampered genomics development, as is the case for several plants of high agronomic value. Isolating single chromosomes or chromosome arms via flow sorting offers a clue to resolve such complexity by focusing sequencing to a discrete and self-consistent part of the whole genome. The occurrence of sufficient differences in the size and or base-pair composition of the individual chromosomes, which is uncommon in plants, is critical for the success of flow sorting. We overcome this limitation by developing a robust method for labeling isolated chromosomes, named Fluorescent In situ Hybridization In suspension (FISHIS). FISHIS employs fluorescently labeled synthetic repetitive DNA probes, which are hybridized, in a wash-less procedure, to chromosomes in suspension following DNA alkaline denaturation. All typical A, B and D genomes of wheat, as well as individual chromosomes from pasta (T. durum L.) and bread (T. aestivum L.) wheat, were flow-sorted, after FISHIS, at high purity. For the first time in eukaryotes, each individual chromosome of a diploid organism, Dasypyrum villosum (L.) Candargy, was flow-sorted regardless of its size or base-pair related content. FISHIS-based chromosome sorting is a powerful and innovative flow cytogenetic tool which can develop new genomic resources from each plant species, where microsatellite DNA probes are available and high quality chromosome suspensions could be produced. The joining of FISHIS labeling and flow sorting with the Next Generation Sequencing methodology will enforce genomics for more species, and by this mightier chromosome approach it will be possible to increase our knowledge about structure, evolution and function of plant genome to be used for crop improvement. It is also anticipated that this technique could contribute to analyze and sort animal chromosomes with peculiar cytogenetic abnormalities, such as copy number variations or cytogenetic aberrations.
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