James Simmonds

A Genetic Framework for Grain Size and Shape Variation in Wheat

Using large-scale quantitative analysis, this work reveals that grain shape and size are independent traits in both modern and primitive wheat and are under the control of distinct genetic components. Moreover, the phenotypic diversity in grain morphology found in modern commercial wheat is the result of a recent and severe bottleneck. Grain morphology in wheat (Triticum aestivum) has been selected and manipulated even in very early agrarian societies and remains a major breeding target. We undertook a large-scale quantitative analysis to determine the genetic basis of the phenotypic diversity in wheat grain morphology. A high-throughput method was used to capture grain size and shape variation in multiple mapping populations, elite varieties, and a broad collection of ancestral wheat species. This analysis reveals that grain size and shape are largely independent traits in both primitive wheat and in modern varieties. This phenotypic structure was retained across the mapping populations studied, suggesting that these traits are under the control of a limited number of discrete genetic components. We identified the underlying genes as quantitative trait loci that are distinct for grain size and shape and are largely shared between the different mapping populations. Moreover, our results show a significant reduction of phenotypic variation in grain shape in the modern germplasm pool compared with the ancestral wheat species, probably as a result of a relatively recent bottleneck. Therefore, this study provides the genetic underpinnings of an emerging phenotypic model where wheat domestication has transformed a long thin primitive grain to a wider and shorter modern grain.

Uncovering hidden variation in polyploid wheat

Significance Pasta and bread wheat are polyploid species that carry multiple copies of each gene. Therefore, loss-of-function mutations in one gene copy are frequently masked by functional copies on other genomes. We sequenced the protein coding regions of 2,735 mutant lines and developed a public database including more than 10 million mutations. Researchers and breeders can search this database online, identify mutations in the different copies of their target gene, and request seeds to study gene function or improve wheat varieties. Mutations are being used to improve the nutritional value of wheat, increase the size of the wheat grains, and generate additional variability in flowering genes to improve wheat adaptation to new and changing environments. Comprehensive reverse genetic resources, which have been key to understanding gene function in diploid model organisms, are missing in many polyploid crops. Young polyploid species such as wheat, which was domesticated less than 10,000 y ago, have high levels of sequence identity among subgenomes that mask the effects of recessive alleles. Such redundancy reduces the probability of selection of favorable mutations during natural or human selection, but also allows wheat to tolerate high densities of induced mutations. Here we exploited this property to sequence and catalog more than 10 million mutations in the protein-coding regions of 2,735 mutant lines of tetraploid and hexaploid wheat. We detected, on average, 2,705 and 5,351 mutations per tetraploid and hexaploid line, respectively, which resulted in 35–40 mutations per kb in each population. With these mutation densities, we identified an average of 23–24 missense and truncation alleles per gene, with at least one truncation or deleterious missense mutation in more than 90% of the captured wheat genes per population. This public collection of mutant seed stocks and sequence data enables rapid identification of mutations in the different copies of the wheat genes, which can be combined to uncover previously hidden variation. Polyploidy is a central phenomenon in plant evolution, and many crop species have undergone recent genome duplication events. Therefore, the general strategy and methods developed herein can benefit other polyploid crops.

Susceptibility to Fusarium head blight is associated with the Rht-D1b semi-dwarfing allele in wheat

Fusarium head blight (FHB) is an important disease of wheat worldwide. The cultivar Spark is more resistant than most other UK winter wheat varieties but the genetic basis for this is not known. A mapping population from a cross between Spark and the FHB susceptible variety Rialto was used to identify quantitative trait loci (QTL) associated with resistance. QTL analysis across environments revealed nine QTL for FHB resistance and four QTL for plant height (PH). One FHB QTL was coincident with the Rht-1D locus and accounted for up to 51% of the phenotypic variance. The enhanced FHB susceptibility associated with Rht-D1b is not an effect of PH per se as other QTL for height segregating in this population have no influence on susceptibility. Experiments with near-isogenic lines supported the association between susceptibility and the Rht-D1b allele conferring the semi-dwarf habit. Our results demonstrate that lines carrying the Rht-1Db semi-dwarfing allele are compromised in resistance to initial infection (type I resistance) while being unaffected in resistance to spread within the spike (type II resistance).

The Inhibitor of wax 1 locus (Iw1) prevents formation of β‐ and OH‐β‐diketones in wheat cuticular waxes and maps to a sub‐cM interval on chromosome arm 2BS

Glaucousness is described as the scattering effect of visible light from wax deposited on the cuticle of plant aerial organs. In wheat, two dominant genes lead to non-glaucous phenotypes: Inhibitor of wax 1 (Iw1) and Iw2. The molecular mechanisms and the exact extent (beyond visual assessment) by which these genes affect the composition and quantity of cuticular wax is unclear. To describe the Iw1 locus we used a genetic approach with detailed biochemical characterization of wax compounds. Using synteny and a large number of F2 gametes, Iw1 was fine-mapped to a sub-cM genetic interval on wheat chromosome arm 2BS, which includes a single collinear gene from the corresponding Brachypodium and rice physical maps. The major components of flag leaf and peduncle cuticular waxes included primary alcohols, β-diketones and n-alkanes. Small amounts of C19-C27 alkyl and methylalkylresorcinols that have not previously been described in wheat waxes were identified. Using six pairs of BC2 F3 near-isogenic lines, we show that Iw1 inhibits the formation of β- and hydroxy-β-diketones in the peduncle and flag leaf blade cuticles. This inhibitory effect is independent of genetic background or tissue, and is accompanied by minor but consistent increases in n-alkanes and C24 primary alcohols. No differences were found in cuticle thickness and carbon isotope discrimination in near-isogenic lines differing at Iw1.

Identification and independent validation of a stable yield and thousand grain weight QTL on chromosome 6A of hexaploid wheat ( Triticum aestivum L .)

BackgroundGrain yield in wheat is a polygenic trait that is influenced by environmental and genetic interactions at all stages of the plant’s growth. Yield is usually broken down into three components; number of spikes per area, grain number per spike, and grain weight (TGW). In polyploid wheat, studies have identified quantitative trait loci (QTL) which affect TGW, yet few have been validated and fine-mapped using independent germplasm, thereby having limited impact in breeding.ResultsIn this study we identified a major QTL for TGW, yield and green canopy duration on wheat chromosome 6A of the Spark x Rialto population, across 12 North European environments. Using independent germplasm in the form of BC2 and BC4 near isogenic lines (NILs), we validated the three QTL effects across environments. In four of the five experiments the Rialto 6A introgression gave significant improvements in yield (5.5%) and TGW (5.1%), with morphometric measurements showing that the increased grain weight was a result of wider grains. The extended green canopy duration associated with the high yielding/TGW Rialto allele was comprised of two independent effects; earlier flowering and delayed final maturity, and was expressed stably across the five environments. The wheat homologue (TaGW2) of a rice gene associated with increased TGW and grain width was mapped within the QTL interval. However, no polymorphisms were identified in the coding sequence between the parents.ConclusionThe discovery and validation through near-isogenic lines of robust QTL which affect yield, green canopy duration, thousand grain weight, and grain width on chromosome 6A of hexaploid wheat provide an important first step to advance our understanding of the genetic mechanisms regulating the complex processes governing grain size and yield in polyploid wheat.

Speed breeding is a powerful tool to accelerate crop research and breeding

The growing human population and a changing environment have raised significant concern for global food security, with the current improvement rate of several important crops inadequate to meet future demand1. This slow improvement rate is attributed partly to the long generation times of crop plants. Here, we present a method called ‘speed breeding’, which greatly shortens generation time and accelerates breeding and research programmes. Speed breeding can be used to achieve up to 6 generations per year for spring wheat (Triticum aestivum), durum wheat (T. durum), barley (Hordeum vulgare), chickpea (Cicer arietinum) and pea (Pisum sativum), and 4 generations for canola (Brassica napus), instead of 2–3 under normal glasshouse conditions. We demonstrate that speed breeding in fully enclosed, controlled-environment growth chambers can accelerate plant development for research purposes, including phenotyping of adult plant traits, mutant studies and transformation. The use of supplemental lighting in a glasshouse environment allows rapid generation cycling through single seed descent (SSD) and potential for adaptation to larger-scale crop improvement programs. Cost saving through light-emitting diode (LED) supplemental lighting is also outlined. We envisage great potential for integrating speed breeding with other modern crop breeding technologies, including high-throughput genotyping, genome editing and genomic selection, accelerating the rate of crop improvement.Fully enclosed, controlled-environment growth chambers can accelerate plant development. Such ‘speed breeding’ reduces generation times to accelerate crop breeding and research programmes, and can integrate with other modern crop breeding technologies.

Identification of a major QTL controlling the content of B-type starch granules in Aegilops

Starch within the endosperm of most species of the Triticeae has a unique bimodal granule morphology comprising large lenticular A-type granules and smaller near-spherical B-type granules. However, a few wild wheat species (Aegilops) are known to lack B-granules. Ae. peregrina and a synthetic tetraploid Aegilops with the same genome composition (SU) were found to differ in B-granule number. The synthetic tetraploid had normal A- and B-type starch granules whilst Ae. peregrina had only A-granules because the B-granules failed to initiate. A population segregating for B-granule number was generated by crossing these two accessions and was used to study the genetic basis of B-granule initiation. A combination of Bulked Segregant Analysis and QTL mapping identified a major QTL located on the short arm of chromosome 4S that accounted for 44.4% of the phenotypic variation. The lack of B-granules in polyploid Aegilops with diverse genomes suggests that the B-granule locus has been lost several times independently during the evolution of the Triticeae. It is proposed that the B-granule locus is susceptible to silencing during polyploidization and a model is presented to explain the observed data based on the assumption that the initiation of B-granules is controlled by a single major locus per haploid genome.

Genetic dissection of grain size and grain number trade-offs in CIMMYT Wheat Germplasm

Grain weight (GW) and number per unit area of land (GN) are the primary components of grain yield in wheat. In segregating populations both yield components often show a negative correlation among themselves. Here we use a recombinant doubled haploid population of 105 individuals developed from the CIMMYT varieties Weebill and Bacanora to understand the relative contribution of these components to grain yield and their interaction with each other. Weebill was chosen for its high GW and Bacanora for high GN. The population was phenotyped in Mexico, Argentina, Chile and the UK. Two loci influencing grain yield were indicated on 1B and 7B after QTL analysis. Weebill contributed the increasing alleles. The 1B effect, which is probably caused by to the 1BL.1RS rye introgression in Bacanora, was a result of increased GN, whereas, the 7B QTL controls GW. We concluded that increased in GW from Weebill 7B allele is not accompanied by a significant reduction in grain number. The extent of the GW and GN trade-off is reduced. This makes this locus an attractive target for marker assisted selection to develop high yielding bold grain varieties like Weebill. AMMI analysis was used to show that the 7B Weebill allele appears to contribute to yield stability.

Inheritance and QTL mapping of leaf rust resistance in the European winter wheat cultivar ‘Beaver’

Genetic studies were conducted on an European winter wheat cultivar, Beaver, to determine the mode of inheritance of leaf rust resistance at seedling and adult plant growth stages using a recombinant doubled haploid population, Beaver/Soissons. Greenhouse studies indicated the involvement of genes Lr13 and Lr26 in governing leaf rust resistance at seedling growth stages, whereas, adult plant resistance (APR) in the field with pathotypes carrying virulence individually for Lr13 and Lr26 showed trigenic inheritance for the population. Marker regression analysis of adult plant field data indicated the involvement of six significant QTLs (chromosomes 1B, 3B, 3D, 4B, 4D and 5A) in year 2005, four QTLs (1B, 3B, 4B and 5A) in 2006, and six QTLs (1A, 1B, 3B, 4A, 4B and 5A) in 2007 for reducing leaf rust severity. QTLs on chromosomes 1B, 4B and 5A were considered the most important because of their detection across years, whereas QTLs on chromosomes 1A, 3B, 3D and 4A were either inconsistent or non-significant and unexplained. Based on an association of closely linked markers with phenotypic data, putative single gene stocks were identified for each consistent QTL and crossing was initiated to develop populations segregating for each to permit fine mapping of the identified regions.

The wheat Phs-A1 pre-harvest sprouting resistance locus delays the rate of seed dormancy loss and maps 0.3 cM distal to the PM19 genes in UK germplasm

Highlight Phs-A1 confers resistance to sprouting in wheat by delaying the rate of seed dormancy loss and is distinct from the previously proposed PM19 candidate genes.