The wheat Seven in absentia gene is associated with increases in biomass and yield in hot climates.

Wheat (Triticum aestivum L.) productivity is severely reduced by high temperatures. Breeding of heat-tolerant cultivars can be achieved by identifying genes controlling physiological and agronomical traits when high temperatures occur and using these to select superior genotypes, but no gene underlying genetic variation for heat tolerance has previously been described. We advanced the positional cloning of qYDH.3BL, a quantitative trait locus (QTL) on bread wheat chromosome 3B associated with increased yield in hot and dry climates. The delimited genomic region contained 12 putative genes and a sequence variant in the promoter region of one gene, Seven in absentia, TaSINA. This was associated with the QTL's effects on early vigour, root growth, plant biomass, and yield components in two distinct wheat populations grown under various growth conditions. Near isogenic lines carrying the positive allele at qYDH.3BL underexpressed TaSINA and had increased vigour and water use efficiency early in development, as well as increased biomass, grain number, and grain weight following heat stress. A survey of worldwide distribution indicated that the positive allele became widespread from the 1950s through the CIMMYT wheat breeding programme but, to date, has been selected only in breeding programmes in Mexico and Australia.

Development of an Australian Bread Wheat Nested Association Mapping Population, a New Genetic Diversity Resource for Breeding under Dry and Hot Climates.

Genetic diversity, knowledge of the genetic architecture of the traits of interest and efficient means of transferring the desired genetic diversity into the relevant genetic background are prerequisites for plant breeding. Exotic germplasm is a rich source of genetic diversity; however, they harbor undesirable traits that limit their suitability for modern agriculture. Nested association mapping (NAM) populations are valuable genetic resources that enable incorporation of genetic diversity, dissection of complex traits and providing germplasm to breeding programs. We developed the OzNAM by crossing and backcrossing 73 diverse exotic parents to two Australian elite varieties Gladius and Scout. The NAM parents were genotyped using the iSelect wheat 90K Infinium SNP array, and the progeny were genotyped using a custom targeted genotyping-by-sequencing assay based on molecular inversion probes designed to target 12,179 SNPs chosen from the iSelect wheat 90K Infinium SNP array of the parents. In total, 3535 BC1F4:6 RILs from 125 families with 21 to 76 lines per family were genotyped and we found 4964 polymorphic and multi-allelic haplotype markers that spanned the whole genome. A subset of 530 lines from 28 families were evaluated in multi-environment trials over three years. To demonstrate the utility of the population in QTL mapping, we chose to map QTL for maturity and plant height using the RTM-GWAS approach and we identified novel and known QTL for maturity and plant height.

Genetic association of stomatal traits and yield in wheat grown in low rainfall environments.

BACKGROUND: In wheat, grain filling is closely related to flag leaf characteristics and function. Stomata are specialized leaf epidermal cells which regulate photosynthetic CO2 uptake and water loss by transpiration. Understanding the mechanisms controlling stomatal size, and their opening under drought, is critical to reduce plant water loss and maintain a high photosynthetic rate which ultimately leads to elevated yield. We applied a leaf imprinting method for rapid and non-destructive phenotyping to explore genetic variation and identify quantitative traits loci (QTL) for stomatal traits in wheat grown under greenhouse and field conditions. RESULTS: The genetics of stomatal traits on the adaxial surface of the flag leaf was investigated using 146 double haploid lines derived from a cross between two Australian lines of Triticum aestivum, RAC875 and Kukri. The drought tolerant line RAC875 showed numerous small stomata in contrast to Kukri. Significant differences between the lines were observed for stomatal densitity and size related traits. A negative correlation was found between stomatal size and density, reflecting a compensatory relationship between these traits to maintain total pore area per unit leaf surface area. QTL were identified for stomatal traits on chromosomes 1A, 1B, 2B, and 7A under field and controlled conditions. Most importantly some of these loci overlap with QTL on chromosome 7A that control kernel number per spike, normalized difference vegetation index, harvest index and yield in the same population. CONCLUSIONS: In this first study to decifer genetic relationships between wheat stomatal traits and yield in response to water deficit, no significant correlations were observed among yield and stomatal traits under field conditions. However we found some overlaps between QTL for stomatal traits and yield across environments. This suggested that stomatal traits could be an underlying mechanism increasing yield at specific loci and used as a proxy to track a target QTL in recombinant lines. This finding is a step-forward in understanding the function of these loci and identifying candidate genes to accelerate positional cloning of yield QTL in wheat under drought.

Transcripts of wheat at a target locus on chromosome 6B associated with increased yield, leaf mass and chlorophyll index under combined drought and heat stress.

Drought and heat stress constrain wheat (Triticum aestivum L.) yields globally. To identify putative mechanisms and candidate genes associated with combined drought and heat stress tolerance, we developed bread wheat near-isogenic lines (NILs) targeting a quantitative trait locus (QTL) on chromosome 6B which was previously associated with combined drought and heat stress tolerance in a diverse panel of wheats. Genotyping-by-sequencing was used to identify additional regions that segregated in allelic pairs between the recurrent and the introduced exotic parent, genome-wide. NILs were phenotyped in a gravimetric platform with precision irrigation and exposed to either drought or to combined drought and heat stress from three days after anthesis. An increase in grain weight in NILs carrying the exotic allele at 6B locus was associated with thicker, greener leaves, higher photosynthetic capacity and increased water use index after re-watering. RNA sequencing of developing grains at early and later stages of treatment revealed 75 genes that were differentially expressed between NILs across both treatments and timepoints. Differentially expressed genes coincided with the targeted QTL on chromosome 6B and regions of genetic segregation on chromosomes 1B and 7A. Pathway enrichment analysis showed the involvement of these genes in cell and gene regulation, metabolism of amino acids and transport of carbohydrates. The majority of these genes have not been characterized previously under drought or heat stress and they might serve as candidate genes for improved abiotic stress tolerance.

Novel Alleles for Combined Drought and Heat Stress Tolerance in Wheat.

Drought and heat waves commonly co-occur in many wheat-growing regions causing significant crop losses. The identification of stress associated quantitative trait loci, particularly those for yield, is problematic due to their association with plant phenology and the high genetic × environment interaction. Here we studied a panel of 315 diverse, spring type accessions of bread wheat (Triticum aestivum) in pots in a semi-controlled environment under combined drought and heat stress over 2 years. Importantly, we treated individual plants according to their flowering time. We found 134 out of the 145 identified loci for grain weight that were not associated with either plant phenology or plant height. The majority of loci uncovered here were novel, with favorable alleles widespread in Asian and African landraces providing opportunities for their incorporation into modern varieties through breeding. Using residual heterozygosity in lines from a nested association mapping population, we were able to rapidly develop near-isogenic lines for important target loci. One target locus on chromosome 6A contributed to higher grain weight, harvest index, thousand kernel weight, and grain number under drought and heat stress in field conditions consistent with allelic effects demonstrated in the genome-wide association study.

Novel Informatic Tools to Support Functional Annotation of the Durum Wheat Genome.

Seed mutagenesis is one strategy to create a population with thousands of useful mutations for the direct selection of desirable traits, to introduce diversity into varietal improvement programs, or to generate a mutant collection to support gene functional analysis. However, phenotyping such large collections, where each individual may carry many mutations, is a bottleneck for downstream analysis. Targeting Induced Local Lesions in Genomes (TILLinG), when coupled with next-generation sequencing allows high-throughput mutation discovery and selection by genotyping. We mutagenized an advanced durum breeding line, UAD0951096_F2:5 and performed short-read (2x125 bp) Illumina sequencing of the exome of 100 lines using an available exome capture platform. To improve variant calling, we generated a consolidated exome reference using the recently available genome sequences of the cultivars Svevo and Kronos to facilitate the alignment of reads from the UAD0951096_F2:5 derived mutants. The resulting exome reference was 484.4 Mbp. We also developed a user-friendly, searchable database and bioinformatic analysis pipeline that allowed us to predict zygosity of the mutations discovered and extracts flanking sequences for rapid marker development. Here, we present these tools with the aim of allowing researchers fast and accurate downstream selection of mutations discovered by TILLinG by sequencing to support functional annotation of the durum wheat genome.