Bread wheat (Triticum aestivum) is a globally important crop, accounting for 20 per cent of the calories consumed by humans. Major efforts are underway worldwide to increase wheat production by extending genetic diversity and analysing key traits, and genomic resources can accelerate progress. But so far the very large size and polyploid complexity of the bread wheat genome have been substantial barriers to genome analysis. Here we report the sequencing of its large, 17-gigabase-pair, hexaploid genome using 454 pyrosequencing, and comparison of this with the sequences of diploid ancestral and progenitor genomes. We identified between 94,000 and 96,000 genes, and assigned two-thirds to the three component genomes (A, B and D) of hexaploid wheat. High-resolution synteny maps identified many small disruptions to conserved gene order. We show that the hexaploid genome is highly dynamic, with significant loss of gene family members on polyploidization and domestication, and an abundance of gene fragments. Several classes of genes involved in energy harvesting, metabolism and growth are among expanded gene families that could be associated with crop productivity. Our analyses, coupled with the identification of extensive genetic variation, provide a resource for accelerating gene discovery and improving this major crop.

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Analysis of the bread wheat genome using whole-genome shotgun sequencing

We report the map-based cloning of the leaf rust resistance gene Lr21, previously mapped to a gene-rich region at the distal end of chromosome arm 1DS of bread wheat (Triticum aestivum L.). Molecular cloning of Lr21 was facilitated by diploid/polyploid shuttle mapping strategy. Cloning of Lr21 was confirmed by genetic transformation and by a stably inherited resistance phenotype in transgenic plants. Lr21 spans 4318 bp and encodes a 1080-amino-acid protein containing a conserved nucleotide-binding site (NBS) domain, 13 imperfect leucine-rich repeats (LRRs), and a unique 151-amino-acid sequence missing from known NBS-LRR proteins at the N terminus. Fine-structure genetic analysis at the Lr21 locus detected a noncrossover (recombination without exchange of flanking markers) within a 1415-bp region resulting from either a gene conversion tract of at least 191 bp or a double crossover. The successful map-based cloning approach as demonstrated here now opens the door for cloning of many crop-specific agronomic traits located in the gene-rich regions of bread wheat.

https://www.genetics.org/content/genetics/164/2/655.full.pdf

Map-based cloning of leaf rust resistance gene Lr21 from the large and polyploid genome of bread wheat.

One hundred and seventy-eight loci have been mapped in Triticum tauschii (Coss.) Schmal. (2n = 14, DD) and Triticum aestivum L. em. Thell. (2n = 42, AABBDD). Thirty-five loci were mapped by aneuplo...

A genetic linkage map of Triticum tauschii (DD) and its relationship to the D genome of bread wheat (AABBDD)

To improve our understanding of the organization and evolution of the wheat (Triticum aestivum) genome, we sequenced and annotated 13-Mb contigs (18.2 Mb) originating from different regions of its largest chromosome, 3B (1 Gb), and produced a 2x chromosome survey by shotgun Illumina/Solexa sequencing. All regions carried genes irrespective of their chromosomal location. However, gene distribution was not random, with 75% of them clustered into small islands containing three genes on average. A twofold increase of gene density was observed toward the telomeres likely due to high tandem and interchromosomal duplication events. A total of 3222 transposable elements were identified, including 800 new families. Most of them are complete but showed a highly nested structure spread over distances as large as 200 kb. A succession of amplification waves involving different transposable element families led to contrasted sequence compositions between the proximal and distal regions. Finally, with an estimate of 50,000 genes per diploid genome, our data suggest that wheat may have a higher gene number than other cereals. Indeed, comparisons with rice (Oryza sativa) and Brachypodium revealed that a high number of additional noncollinear genes are interspersed within a highly conserved ancestral grass gene backbone, supporting the idea of an accelerated evolution in the Triticeae lineages.

/pdf/megabase-level-sequencing-reveals-contrasted-organization-zzfhx3djjs.pdf

Megabase Level Sequencing Reveals Contrasted Organization and Evolution Patterns of the Wheat Gene and Transposable Element Spaces

Arabidopsis thaliana is a small flowering plant with various properties that make it an excellent organism for experiments in molecular genetics. These properties include having a small nuclear genome, a near absence of dispersed repetitive DNA, and a generation time of 4 to 5 weeks. In addition, mutations that affect hormone synthesis and response, many different enzyme activities, and numerous developmental processes have been identified and characterized.

https://science.sciencemag.org/content/sci/229/4719/1214.full.pdf

Arabidopsis thaliana and Plant Molecular Genetics.

Reassociation kinetics of sheared hexaploid wheat DNA in 0.18 M Na+ at 60 °C show the presence of three major classes of nucleotide sequences; (1) a very rapidly reannealing fracton that comprises about 4 to 10% of the genome and reanneals virtually instantly. This fraction contains sequences which appear to be randomly distributed through at least 40% of the genome; they may be palindromic sequences; (2) a heterogeneous intermediate reannealing fraction comprising about 70–80% of the genome which consists of families with apparent repetition frequencies ranging from about 100–100000; and (3) a slow reannealing fraction. These fractions have been isolated and studied separately. Seventy percent of the slow reannealing fraction (12 to 20% of the genome) was found to contain sequences present in approximately six copies per hexaploid genome. The single copy sequences in the three constituent diploid genomes of hexaploid wheat appear to show near complete homology to one another.

D. B. Smith

Papers

Characterisation of the wheat genome by renaturation kinetics