Pyramiding wheat pre-harvest sprouting resistance genes in triticale breeding.

Pre -harvest sprouting (PHS) is an important problem in cereal production reducing yield and grain quality. After decades of improvement, triticale remains particularly susceptible to PHS but no resistance genes or QTLs were identified so far in this species. As wheat shares the A and B genomes with triticale, wheat PHS resistance genes can be introgressed into triticale genome by recombination after interspecific crosses. In this project, three PHS resistance genes have been transferred from wheat to triticale by marker-assisted interspecific crosses, followed by four backcrosses. The gene TaPHS1 from the 3AS chromosome of cultivar Zenkoujikomugi (Zen) and the TaMKK3 and TaQsd1, respectively located on the 4AL and 5BL chromosomes derived both from cultivar Aus1408, were pyramided in the triticale cultivar Cosinus. Only the TaPHS1 gene increases consistently the PHS resistance in triticale. The lack of efficacy of the other two genes, especially TaQsd1, could be the result of an imperfect linkage between the marker and the gene of interest. The introduction of PHS resistance genes did not alter agronomic nor disease resistance performances of triticale. This approach leads to two new, agronomically performant and PHS-resistant triticale cultivars. Today, two breeding triticale lines are ready to enter the official registration process.

Rapid cloning of genes in hexaploid wheat using cultivar-specific long-range chromosome assembly.

Cereal crops such as wheat and maize have large repeat-rich genomes that make cloning of individual genes challenging. Moreover, gene order and gene sequences often differ substantially between cultivars of the same crop species. A major bottleneck for gene cloning in cereals is the generation of high-quality sequence information from a cultivar of interest. In order to accelerate gene cloning from any cropping line, we report 'targeted chromosome-based cloning via long-range assembly' (TACCA). TACCA combines lossless genome-complexity reduction via chromosome flow sorting with Chicago long-range linkage to assemble complex genomes. We applied TACCA to produce a high-quality (N50 of 9.76 Mb) de novo chromosome assembly of the wheat line CH Campala Lr22a in only 4 months. Using this assembly we cloned the broad-spectrum Lr22a leaf-rust resistance gene, using molecular marker information and ethyl methanesulfonate (EMS) mutants, and found that Lr22a encodes an intracellular immune receptor homologous to the Arabidopsis thaliana RPM1 protein.

The cre1 and cre3 nematode resistance genes are located at homeologous loci in the wheat genome.

Differential responses in host-nematode pathotype interactions occur in wheat lines carrying different cereal cyst nematode resistance (Cre) genes. Cre1, located on chromosome 2B, confers resistance to most European nematodes and the sole Australian pathotype, while Cre3, present on chromosome 2D, is highly resistant to the Australian pathotype and susceptible to a number of European pathotypes. Genes encoding nucleotide binding site-leucine rich repeat (NBS-LRR) proteins that cosegregate with the Cre3 locus cross hybridize to homologues whose restriction fragment length polymorphism (RFLP) patterns distinguish near-isogenic Cre1 nematode-resistant wheat lines. Genetic mapping showed that the NBS-LRR gene members that distinguished the Cre1 near-isogenic lines were located on chromosome 2BL at a locus, designated Xcsl107, that cosegregates with the Cre1 locus. A haplotype of NBS-LRR genes from the Xcsl107 locus provides a diagnostic marker for the presence of Cre1 nematode resistance in a wide collection of wheat lines and segregating families. Genetic analysis of NBS-LRR haplotypes that cosegregate with Cre1 and Cre3 resistance, together with flanking cDNA markers and other markers from homoeologous group 2 chromosomes, revealed a conserved gene order that suggests Cre1 and Cre3 are homeoloci.

Surveying of pollen-mediated crop-to-crop gene flow from a wheat field trial as a biosafety measure.

Outcrosses from genetically modified (GM) to conventional crops by pollen-mediated gene flow (PMGF) are a concern when growing GM crops close to non-GM fields. This also applies to the experimental releases of GM plants in field trials. Therefore, biosafety measures such as isolation distances and surveying of PMGF are required by the regulatory authorities in Switzerland. For two and three years, respectively, we monitored crop-to-crop PMGF from GM wheat field trials in two locations in Switzerland. The pollen donors were two GM spring wheat lines with enhanced fungal resistance and a herbicide tolerance as a selection marker. Seeds from the experimental plots were sampled to test the detection method for outcrosses. Two outcrosses were found adjacent to a transgenic plot within the experimental area. For the survey of PMGF, pollen receptor plots of the conventional wheat variety Frisal used for transformation were planted in the border crop and around the experimental field up to a distance of 200 m. Although the environmental conditions were favorable and the donor and receptor plots flowered at the same time, only three outcrosses were found in approximately 185,000 tested seedlings from seeds collected outside the experimental area. All three hybrids were found in the border crop surrounding the experimental area, but none outside the field. We conclude that a pollen barrier (border crop) and an additional isolation distance of 5 m is a sufficient measure to reduce PMGF from a GM wheat field trial to cleistogamous varieties in commercial fields below a level that can be detected.