573 research outputs found
Characterisation of Perennial Ryegrass Parental Inbred Lines for Generating Recombinant Inbred Lines for Fine Mapping and Gene Cloning
Intermated recombinant inbred lines (IRIs) are a powerful tool for fine mapping and cloning of genes. Such population structures have been particularly helpful for cloning of genes in the model genetic plant Arabidopsis thaliana. IRIs or recombinant inbred lines (RILs) would be valuable for perennial ryegrass (Lolium perenne), but its allogamous character means that the construction of RILs is a difficult task. The international Lolium community would benefit from the development of such lines. The aims of the projects are to characterise the parental lines and initial generations at the (1) phenotypic, (2) molecular and (3) molecular cytogenetic level
Introgression of chromosome segments from multiple alien species in wheat breeding lines with wheat streak mosaic virus resistance
Pyramiding of alien-derived Wheat streak mosaic virus (WSMV) resistance and resistance enhancing genes in wheat is a costeffective and environmentally safe strategy for disease control. PCR-based markers and cytogenetic analysis with genomic in situ hybridisation were applied to identify alien chromatin in four genetically diverse populations of wheat (Triticum aestivum) lines incorporating chromosome segments from Thinopyrum intermedium and Secale cereale (rye). Out of 20 experimental lines, 10 carried Th. intermedium chromatin as T4DL*4Ai#2S translocations, while, unexpectedly, 7 lines were positive for alien chromatin (Th. intermedium or rye) on chromosome 1B. The newly described rye 1RS chromatin, transmitted from early in the pedigree, was associated with enhanced WSMV resistance. Under field conditions, the 1RS chromatin alone showed some resistance, while together with the Th. intermedium 4Ai#2S offered superior resistance to that demonstrated by the known resistant cultivar Mace. Most alien wheat lines carry whole chromosome arms, and it is notable that these lines showed intra-arm recombination within the 1BS arm. The translocation breakpoints between 1BS and alien chromatin fell in three categories: (i) at or near to the centromere, (ii) intercalary between markers UL-Thin5 and Xgwm1130 and (iii) towards the telomere between Xgwm0911 and Xbarc194. Labelled genomic Th. intermedium DNA hybridised to the rye 1RS chromatin under high stringency conditions, indicating the presence of shared tandem repeats among the cereals. The novel small alien fragments may explain the difficulty in developing well-adapted lines carrying Wsm1 despite improved tolerance to the virus. The results will facilitate directed chromosome engineering producing agronomically desirable WSMV-resistant germplasm
Diverged subpopulations in tropical Urochloa (Brachiaria) forage species indicate a role for facultative apomixis and varying ploidy in their population structure and evolution
Abstract
Background
Urochloa (syn. Brachiaria) is a genus of tropical grasses sown as forage feedstock, particularly in marginal soils. Here we aimed to clarify the genetic diversity and population structure in Urochloa species to understand better how population evolution relates to ploidy level and occurrence of apomictic reproduction.
Methods
We explored the genetic diversity of 111 accessions from the five Urochloa species used to develop commercial cultivars. These accessions were conserved from wild materials collected at their centre of origin in Africa, and they tentatively represent the complete Urochloa gene pool used in breeding programmes. We used RNA-sequencing to generate 1.1 million single nucleotide polymorphism loci. We employed genetic admixture, principal component and phylogenetic analyses to define subpopulations.
Results
We observed three highly differentiated subpopulations in U. brizantha, which were unrelated to ploidy: one intermixed with U. decumbens, and two diverged from the former and the other species in the complex. We also observed two subpopulations in U. humidicola, unrelated to ploidy; one subpopulation had fewer accessions but included the only characterized sexual accession in the species. Our results also supported a division of U. decumbens between diploids and polyploids, and no subpopulations within U. ruziziensis and U. maxima.
Conclusions
Polyploid U. decumbens are more closely related to polyploid U. brizantha than to diploid U. decumbens, which supports the divergence of both polyploid groups from a common tetraploid ancestor and provides evidence for the hybridization barrier of ploidy. The three differentiated subpopulations of apomictic polyploid U. brizantha accessions constitute diverged ecotypes, which can probably be utilized in hybrid breeding. Subpopulations were not observed in non-apomictic U. ruziziensis. Sexual Urochloa polyploids were not found (U. brizantha, U. decumbens) or were limited to small subpopulations (U. humidicola). The subpopulation structure observed in the Urochloa sexual–apomictic multiploidy complexes supports geographical parthenogenesis, where the polyploid genotypes exploit the evolutionary advantage of apomixis, i.e. uniparental reproduction and clonality, to occupy extensive geographical areas
Complex polyploid and hybrid species in an apomictic and sexual tropical forage grass group: genomic composition and evolution in Urochloa (Brachiaria) species
Background and Aims
Diploid and polyploid Urochloa (including Brachiaria, Panicum and Megathyrsus species) C-4 tropical forage grasses originating from Africa are important for food security and the environment, often being planted in marginal lands worldwide. We aimed to characterize the nature of their genomes, the repetitive DNA and the genome composition of polyploids, leading to a model of the evolutionary pathways within the group including many apomictic species.
Methods
Some 362 forage grass accessions from international germplasm collections were studied, and ploidy was determined using an optimized flow cytometry method. Whole-genome survey sequencing and molecular cytogenetic analysis were used to identify chromosomes and genomes in Urochloa accessions belonging to the 'brizantha' and 'humidicola' agamic complexes and U. maxima.
Key Results
Genome structures are complex and variable, with multiple ploidies and genome compositions within the species, and no clear geographical patterns. Sequence analysis of nine diploid and polyploid accessions enabled identification of abundant genome-specific repetitive DNA motifs. In situ hybridization with a combination of repetitive DNA and genomic DNA probes identified evolutionary divergence and allowed us to discriminate the different genomes present in polyploids.
Conclusions
We suggest a new coherent nomenclature for the genomes present. We develop a model of evolution at the whole-genome level in diploid and polyploid accessions showing processes of grass evolution. We support the retention of narrow species concepts for Urochloa brizantha, U. decumbens and U. ruziziensis, and do not consider diploids and polyploids of single species as cytotypes. The results and model will be valuable in making rational choices of parents for new hybrids, assist in use of the germplasm for breeding and selection of Urochloa with improved sustainability and agronomic potential, and assist in measuring and conserving biodiversity in grasslands
Cytomolecular identification of individual wheat-wheat chromosome arm associations in wheat-rye hybrids
Chromosome pairing in the meiotic metaphase I of wheatrye
hybrids has been characterized by sequential genomic
and fluorescent in situ hybridization allowing not only the
discrimination of wheat and rye chromosomes, but also the
identification of the individual wheat and rye chromosome
arms involved in the chromosome associations. The majority
of associations (93.8%) were observed between the wheat
chromosomes. The largest number of wheat-wheat chromosome
associations (53%) was detected between the A and D
genomes, while the frequency of B-D and A-B associations
was significantly lower (32 and 8%, respectively). Among the
A-D chromosome associations, pairing between the 3AL and
3DL arms was observed with the highest frequency, while
the most frequent of all the chromosome associations (0.113/
cell) was found to be the 3DS-3BS. Differences in the pairing
frequency of the individual chromosome arms of wheat-rye
hybrids have been discussed in relation to the homoeologous
relationships between the constituent genomes of
hexaploid wheat
Application of a modified drop method for high-resolution pachytene chromosome spreads in two Phalaenopsis species
Epigenetic marks in the mature pollen of Quercus suber L. (Fagaceae)
We have analysed the distribution of epigenetic
marks for histone modifications at lysine residues H3 and
H4, and DNA methylation, in the nuclei of mature pollen
cells of the Angiosperm tree Quercus suber; a monoecious
wind pollinated species with a protandrous system, and a
long post-pollination period. The ultrasonic treatment
developed for the isolation of pollen nuclei proved to be a
fast and reliable method, preventing the interference of cell
wall autofluorescence in the in situ immunolabelling
assays. In contrast with previous studies on herbaceous
species with short progamic phases, our results are consistent
with a high level of silent (5-mC and H3K9me2)
epigenetic marks on chromatin of the generative nucleus,
and the prevalence of active marks (H3K9me3 and H4Kac)
in the vegetative nucleus. The findings are discussed in
terms of the pollination/fertilization timing strategy adopted
by this plant specie
Maintenance and expansion of genetic and trait variation following domestication in a clonal crop
Aegilops-Secale amphiploids: chromosome categorisation, pollen viability and identification of fungal disease resistance genes
The aim of this study was to assess the potential breeding value of goatgrass-rye amphiploids, which we are using as a “bridge” in a transfer of Aegilops chromatin (containing, e.g. leaf rust resistance genes) into triticale. We analysed the chromosomal constitution (by genomic in situ hybridisation, GISH), fertility (by pollen viability tests) and the presence of leaf rust and eyespot resistance genes (by molecular and endopeptidase assays) in a collection of 6× and 4× amphiploids originating from crosses between five Aegilops species and Secale cereale. In the five hexaploid amphiploids Aegilops kotschyi × Secale cereale (genome UUSSRR), Ae. variabilis × S. cereale (UUSSRR), Ae. biuncialis × S. cereale (UUMMRR; two lines) and Ae. ovata × S. cereale (UUMMRR), 28 Aegilops chromosomes were recognised, while in the Ae. tauschii × S. cereale amphiploid (4×; DDRR), only 14 such chromosomes were identified. In the materials, the number of rye chromosomes varied from 14 to 16. In one line of Ae. ovata × S. cereale, the U-R translocation was found. Pollen viability varied from 24.4 to 75.4%. The leaf rust resistance genes Lr22, Lr39 and Lr41 were identified in Ae. tauschii and the 4× amphiploid Ae. tauschii × S. cereale. For the first time, the leaf rust resistance gene Lr37 was found in Ae. kotschyi, Ae. ovata, Ae. biuncialis and amphiploids derived from those parental species. No eyespot resistance gene Pch1 was found in the amphiploids
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