BackgroundRye (Secale cereale L.) belongs to tribe Triticeae and is an important temperate cereal. It is one of the parents of man-made species Triticale and has been used as a source of agronomically important genes for wheat improvement. The short arm of rye chromosome 1 (1RS), in particular is rich in useful genes, and as it may increase yield, protein content and resistance to biotic and abiotic stress, it has been introgressed into wheat as the 1BL.1RS translocation. A better knowledge of the rye genome could facilitate rye improvement and increase the efficiency of utilizing rye genes in wheat breeding.ResultsHere, we report on BAC end sequencing of 1,536 clones from two 1RS-specific BAC libraries. We obtained 2,778 (90.4%) useful sequences with a cumulative length of 2,032,538 bp and an average read length of 732 bp. These sequences represent 0.5% of 1RS arm. The GC content of the sequenced fraction of 1RS is 45.9%, and at least 84% of the 1RS arm consists of repetitive DNA. We identified transposable element junctions in BESs and developed insertion site based polymorphism markers (ISBP). Out of the 64 primer pairs tested, 17 (26.6%) were specific for 1RS. We also identified BESs carrying microsatellites suitable for development of 1RS-specific SSR markers.ConclusionThis work demonstrates the utility of chromosome arm-specific BAC libraries for targeted analysis of large Triticeae genomes and provides new sequence data from the rye genome and molecular markers for the short arm of rye chromosome 1.
Crossing over-based recombination is a powerful tool for generating new allelic combinations during sexual reproduction. It usually occurs between homologous chromosomes. However, under some conditions, homoeologues may also be capable of crossing over. Whether homologous and homoeologous crossovers are equivalent and governed by the same rules has never been established. Here we report on chromosome distribution of homoeologous crossovers in a unique system of Festuca × Lolium hybrids. Unlike in most other hybrids, in these intergeneric hybrids, homoeologous chromosomes are capable of pairing and crossing over with frequencies approaching that of homologues. At the same time, genome divergence makes cytological detection of chromosome recombination feasible. We analyzed the distribution of homoeologous recombination along individual chromosomes in a complete set of intergeneric single chromosome substitutions fromF. pratensisinto tetraploid L. multiflorum. Homoeologous recombination sites were not evenly distributed along the chromosomes, being concentrated in intercalary regions of the arms and reduced in proximal and distal regions. Several recombination hotspots and cold spots were found along individual chromosomes and the recombination was not affected by the presence of a secondary constriction. Our results indicate that despite the uneven distribution of homoeologous recombination, introgression of any part of the F. pratensis genome into L. multiflorum is feasible.
SummaryWild emmer wheat (Triticum turgidum ssp. dicoccoides) is the progenitor of wheat. We performed chromosome‐based survey sequencing of the 14 chromosomes, examining repetitive sequences, protein‐coding genes, miRNA/target pairs and tRNA genes, as well as syntenic relationships with related grasses. We found considerable differences in the content and distribution of repetitive sequences between the A and B subgenomes. The gene contents of individual chromosomes varied widely, not necessarily correlating with chromosome size. We catalogued candidate agronomically important loci, along with new alleles and flanking sequences that can be used to design exome sequencing. Syntenic relationships and virtual gene orders revealed several small‐scale evolutionary rearrangements, in addition to providing evidence for the 4AL‐5AL‐7BS translocation in wild emmer wheat. Chromosome‐based sequence assemblies contained five novel miRNA families, among 59 families putatively encoded in the entire genome which provide insight into the domestication of wheat and an overview of the genome content and organization.
Analyses of large genomes are hampered by high proportions of repetitive DNA, that make the assembly of short sequence reads difficult. This is also the case for meadow fescue (Festuca pratensis Huds.), one of predominant grass species in temperate and Northern regions with the genome size estimated at 1C = 3,175 Mbp. This species is known for its ability to survive under freezing conditions and it has been used widely in intergeneric hybridization with various ryegrass species to produce superior Festulolium cultivars. Here we describe attempts to dissect the meadow fescue's genome into smaller fractions-individual chromosomes and groups of chromosomes. Following the methods of flow cytogenetics developed for legumes and cereals, we have developed a chromosome sorting protocol for grasses and currently we are able to sort F. pratensis chromosome 4 (the largest in the genome) and two groups of three chromosomes each: 2, 3, 7 and 1, 5, 6. As the first step we sequenced chromosome 4 by Illumina with 50x coverage and assembled low copy and genic regions. This facilitated detailed comparative analysis with sequenced genomes of rice, Brachypodium and sorghum and provided the first insight into the genome composition of this species. The possibility to purify chromosome 4 opens the way for a more efficient analysis of genetic loci on this chromosome that control important agronomic traits, such as freezing tolerance. Moreover, purified chromosomes are excellent templates for PCR screening as well as cytogenetic and physical mapping.
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