Most cited article - PubMed ID 28448065
Construction of a map-based reference genome sequence for barley, Hordeum vulgare L
Chromatids of mitotic chromosomes were suggested to coil into a helix in early cytological studies and this assumption was recently supported by chromosome conformation capture (3C) sequencing. Still, direct differential visualization of a condensed chromatin fibre confirming the helical model was lacking. Here, we combined Hi-C analysis of purified metaphase chromosomes, biopolymer modelling and spatial structured illumination microscopy of large fluorescently labeled chromosome segments to reveal the chromonema - a helically-wound, 400 nm thick chromatin thread forming barley mitotic chromatids. Chromatin from adjacent turns of the helix intermingles due to the stochastic positioning of chromatin loops inside the chromonema. Helical turn size varies along chromosome length, correlating with chromatin density. Constraints on the observable dimensions of sister chromatid exchanges further supports the helical chromonema model.
- MeSH
- Chromatids * chemistry MeSH
- Chromatin genetics MeSH
- Chromosomes, Plant MeSH
- Chromosomes MeSH
- Hordeum * cytology MeSH
- Metaphase * MeSH
- Microscopy MeSH
- Sister Chromatid Exchange MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Chromatin MeSH
The wild relatives and progenitors of wheat have been widely used as sources of disease resistance (R) genes. Molecular identification and characterization of these R genes facilitates their manipulation and tracking in breeding programmes. Here, we develop a reference-quality genome assembly of the wild diploid wheat relative Aegilops sharonensis and use positional mapping, mutagenesis, RNA-Seq and transgenesis to identify the stem rust resistance gene Sr62, which has also been transferred to common wheat. This gene encodes a tandem kinase, homologues of which exist across multiple taxa in the plant kingdom. Stable Sr62 transgenic wheat lines show high levels of resistance against diverse isolates of the stem rust pathogen, highlighting the utility of Sr62 for deployment as part of a polygenic stack to maximize the durability of stem rust resistance.
Powdery mildew, a common cereal disease caused by the fungus Blumeria graminis, is a major limiting factor of barley production and genetic resistance is the most appropriate protection against it. To aid the breeding of new cultivars and their marketing, resistance genes can be postulated in homogeneous accessions. Although hybrid cultivars (F1) should be homogeneous, they are often not genetically uniform, especially if more than two genotypes are involved in their seed production or due to undesirable self-pollination, out-crossing and mechanical admixtures. To overcome these problems the accepted method of postulating specific resistance genes based on comparing response type arrays (RTAs) of genetically homogeneous cultivars with RTAs of standard genotypes was substituted by analysing the frequency of response types to clusters of pathogen isolates in segregating F2 generations. This method combines a genetic and phytopathological approach for identifying resistance genes. To assess its applicability six hybrid cultivars were screened and from three to seven with a total of 14 resistance genes were found. Two genes were newly located at the Mla locus and their heritability determined. In addition, three unknown dominant genes were detected. This novel, comprehensive and efficient method to identifying resistance genes in hybrid cultivars can also be applied in other cereals and crops.
- MeSH
- Ascomycota pathogenicity MeSH
- Hordeum genetics growth & development microbiology MeSH
- Quantitative Trait, Heritable MeSH
- Chromosome Mapping MeSH
- Plant Diseases microbiology MeSH
- Disease Resistance * MeSH
- Gene Expression Regulation, Plant MeSH
- Plant Proteins genetics MeSH
- Plant Breeding MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Plant Proteins MeSH
Reference genomes of important cereals, including barley, emmer wheat and bread wheat, were released recently. Their comparison with genome size estimates obtained by flow cytometry indicated that the assemblies represent not more than 88-98% of the complete genome. This work is aimed at identifying the missing parts in two cereal genomes and proposing techniques to make the assemblies more complete. We focused on tandemly organised repetitive sequences, known to be underrepresented in genome assemblies generated from short-read sequence data. Our study found arrays of three tandem repeats with unit sizes of 1242 to 2726 bp present in the bread wheat reference genome generated from short reads. However, this and another wheat genome assembly employing long PacBio reads failed in integrating correctly the 2726-bp repeat in the pseudomolecule context. This suggests that tandem repeats of this size, frequently incorporated in unassigned scaffolds, may contribute to shrinking of pseudomolecules without reducing size of the entire assembly. We demonstrate how this missing information may be added to the pseudomolecules with the aid of nanopore sequencing of individual BAC clones and optical mapping. Using the latter technique, we identified and localised a 470-kb long array of 45S ribosomal DNA absent from the reference genome of barley.
- Keywords
- BAC, barley, bread wheat, genome assembly, optical mapping, ribosomal DNA,
- MeSH
- Chromosomes, Plant genetics MeSH
- Genome, Plant * MeSH
- Hordeum genetics MeSH
- Triticum genetics MeSH
- Tandem Repeat Sequences * MeSH
- Publication type
- Journal Article MeSH
Cereal grasses of the Triticeae tribe have been the major food source in temperate regions since the dawn of agriculture. Their large genomes are characterized by a high content of repetitive elements and large pericentromeric regions that are virtually devoid of meiotic recombination. Here we present a high-quality reference genome assembly for barley (Hordeum vulgare L.). We use chromosome conformation capture mapping to derive the linear order of sequences across the pericentromeric space and to investigate the spatial organization of chromatin in the nucleus at megabase resolution. The composition of genes and repetitive elements differs between distal and proximal regions. Gene family analyses reveal lineage-specific duplications of genes involved in the transport of nutrients to developing seeds and the mobilization of carbohydrates in grains. We demonstrate the importance of the barley reference sequence for breeding by inspecting the genomic partitioning of sequence variation in modern elite germplasm, highlighting regions vulnerable to genetic erosion.
- MeSH
- Cell Nucleus genetics MeSH
- Centromere genetics MeSH
- Chromatin genetics metabolism MeSH
- Chromosomes, Plant genetics MeSH
- Genetic Variation MeSH
- Genome, Plant genetics MeSH
- Genomics MeSH
- Haplotypes genetics MeSH
- Hordeum genetics MeSH
- Chromosome Mapping MeSH
- Meiosis genetics MeSH
- Repetitive Sequences, Nucleic Acid genetics MeSH
- Seeds genetics MeSH
- Chromosomes, Artificial, Bacterial genetics MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Names of Substances
- Chromatin MeSH
