Evaluation of the genetic architecture and the potential of genomics-assisted breeding of quality traits in two large panels of durum wheat.

KEY MESSAGE: New QTL for important quality traits in durum were identified, but for most QTL their effect varies depending on the investigated germplasm. Most of the global durum wheat (Triticum turgidum ssp. durum) production is used for human consumption via pasta and to a lower extent couscous and bulgur. Therefore, durum wheat varieties have to fulfill high demands regarding quality traits. In this study, we evaluated the quality traits protein content, sedimentation volume, falling number, vitreousity and thousand kernel weight in a Central European (CP) and a Southern and Western European panel (SP) with 183 and 159 durum lines, respectively, and investigated their genetic architecture by genome-wide association mapping. Except for protein content, we identified QTL explaining a large proportion of the genotypic variance for different traits. However, most of them were identified only in one panel. Nevertheless, for sedimentation volume a genomic region on chromosome 1B appeared important in both durum panels and a BLAST search against the emmer and bread wheat reference genomes points toward the candidate gene Glu-B3. This was further supported by the protein subunit banding pattern via SDS-PAGE gel electrophoresis. For vitreousity, genomic regions on chromosome 7A explained a larger proportion of the genotypic variance in both panels, whereas one QTL, possibly related to the Pinb-2 locus, also slightly influenced the protein content. Within each panel, high prediction abilities for genomic selection were obtained, which, however, dropped considerably when predicting across both panels. Nevertheless, the across-panel prediction ability was still larger than 0.4 for protein content and sedimentation volume, underlining the potential for genomics-aided durum breeding, if laboratory and logistical facilities are available.

Simultaneous improvement of grain yield and protein content in durum wheat by different phenotypic indices and genomic selection.

KEY MESSAGE: Simultaneous improvement of protein content and grain yield by index selection is possible but its efficiency largely depends on the weighting of the single traits. The genetic architecture of these indices is similar to that of the primary traits. Grain yield and protein content are of major importance in durum wheat breeding, but their negative correlation has hampered their simultaneous improvement. To account for this in wheat breeding, the grain protein deviation (GPD) and the protein yield were proposed as targets for selection. The aim of this work was to investigate the potential of different indices to simultaneously improve grain yield and protein content in durum wheat and to evaluate their genetic architecture towards genomics-assisted breeding. To this end, we investigated two different durum wheat panels comprising 159 and 189 genotypes, which were tested in multiple field locations across Europe and genotyped by a genotyping-by-sequencing approach. The phenotypic analyses revealed significant genetic variances for all traits and heritabilities of the phenotypic indices that were in a similar range as those of grain yield and protein content. The GPD showed a high and positive correlation with protein content, whereas protein yield was highly and positively correlated with grain yield. Thus, selecting for a high GPD would mainly increase the protein content whereas a selection based on protein yield would mainly improve grain yield, but a combination of both indices allows to balance this selection. The genome-wide association mapping revealed a complex genetic architecture for all traits with most QTL having small effects and being detected only in one germplasm set, thus limiting the potential of marker-assisted selection for trait improvement. By contrast, genome-wide prediction appeared promising but its performance strongly depends on the relatedness between training and prediction sets.

Optimum breeding strategies using genomic selection for hybrid breeding in wheat, maize, rye, barley, rice and triticale.

KEY MESSAGE: A breeding strategy with moderate nursery selection followed by genomic selection and one-stage phenotypic selection maximizes annual selection gain for grain yield across a wide range of hybrid breeding scenarios. Genomic selection (GS) is a promising method for the selection of quantitatively inherited traits but its most effective implementation in routine hybrid breeding schemes requires further research. We compared five breeding strategies and varied their available budget, the costs for doubled haploid (DH) line and hybrid seed production as well as variance components for grain yield in a wide range. In contrast to previous studies, we included a nursery selection for disease resistance just before GS on grain yield. The breeding strategy GSrapid with moderate nursery selection followed by one stage GS and one final stage with phenotypic selection on grain yield had the highest annual selection gain across all strategies, budgets, costs and variance components considered and we, therefore, highly recommend its use in hybrid breeding of cereals. Although selecting on traits not correlated with grain yield in the observation nursery, this selection reduced the selection gain of grain yield, especially in the breeding schemes with GS and for selected fractions smaller than 0.3. Owing to the very high number of test candidates entering breeding strategies with GS, the costs for DH line production had a larger impact on the annual selection gain than the hybrid seed production costs. The optimum allocation of test resources maximizing annual selection gain in classical two-stage phenotypic selection on grain yield and for the recommended breeding strategy GSrapid is finally explored for maize, wheat, rye, barley, rice and triticale.

Evaluation of multi-locus models for genome-wide association studies: a case study in sugar beet.

Association mapping has become a widely applied genomic approach to dissect the genetic architecture of complex traits. A major issue for association mapping is the need to control for the confounding effects of population structure, which is commonly done by mixed models incorporating kinship information. In this case study, we employed experimental data from a large sugar beet population to evaluate multi-locus models for association mapping. As in linkage mapping, markers are selected as cofactors to control for population structure and genetic background variation. We compared different biometric models with regard to important quantitative trait locus (QTL) mapping parameters like the false-positive rate, the QTL detection power and the predictive power for the proportion of explained genotypic variance. Employing different approaches we show that the multi-locus model, that is, incorporating cofactors, outperforms the other models, including the mixed model used as a reference model. Thus, multi-locus models are an attractive alternative for association mapping to efficiently detect QTL for knowledge-based breeding.

Cross-validation in association mapping and its relevance for the estimation of QTL parameters of complex traits.

Association mapping has become a widely applied genomic approach to identify quantitative trait loci (QTL) and dissect the genetic architecture of complex traits. However, approaches to assess the quality of the obtained QTL results are lacking. We therefore evaluated the potential of cross-validation in association mapping based on a large sugar beet data set. Our results show that the proportion of the population that should be used as estimation and validation sets, respectively, depends on the size of the mapping population. Generally, a fivefold cross-validation, that is, 20% of the lines as independent validation set, appears appropriate for commonly used population sizes. The predictive power for the proportion of genotypic variance explained by QTL was overestimated by on average 38% indicating a strong bias in the estimated QTL effects. The cross-validated predictive power ranged between 4 and 50%, which are more realistic estimates of this parameter for complex traits. In addition, QTL frequency distributions can be used to assess the precision of QTL position estimates and the robustness of the detected QTL. In summary, cross-validation can be a valuable tool to assess the quality of QTL parameters in association mapping.

Relatedness severely impacts accuracy of marker-assisted selection for disease resistance in hybrid wheat.

The accuracy of genomic selection depends on the relatedness between the members of the set in which marker effects are estimated based on evaluation data and the types for which performance is predicted. Here, we investigate the impact of relatedness on the performance of marker-assisted selection for fungal disease resistance in hybrid wheat. A large and diverse mapping population of 1739 elite European winter wheat inbred lines and hybrids was evaluated for powdery mildew, leaf rust and stripe rust resistance in multi-location field trials and fingerprinted with 9 k and 90 k SNP arrays. Comparison of the accuracies of prediction achieved with data sets from the two marker arrays revealed a crucial role for a sufficiently high marker density in genome-wide association mapping. Cross-validation studies using test sets with varying degrees of relationship to the corresponding estimation sets revealed that close relatedness leads to a substantial increase in the proportion of total genotypic variance explained by the identified QTL and consequently to an overoptimistic judgment of the precision of marker-assisted selection.

Optimum design of family structure and allocation of resources in association mapping with lines from multiple crosses.

Family mapping is based on multiple segregating families and is becoming increasingly popular because of its advantages over population mapping. Athough much progress has been made recently, the optimum design and allocation of resources for family mapping remains unclear. Here, we addressed these issues using a simulation study, resample model averaging and cross-validation approaches. Our results show that in family mapping, the predictive power and the accuracy of quatitative trait loci (QTL) detection depend greatly on the population size and phenotyping intensity. With small population sizes or few test environments, QTL results become unreliable and are hampered by a large bias in the estimation of the proportion of genotypic variance explained by the detected QTL. In addition, we observed that even though good results can be achieved with low marker densities, no plateau is reached with our full marker complement. This suggests that higher quality results could be achieved with greater marker densities or sequence data, which will be available in the near future for many species.

Comparison of biometrical models for joint linkage association mapping.

Joint linkage association mapping (JLAM) combines the advantages of linkage mapping and association mapping, and is a powerful tool to dissect the genetic architecture of complex traits. The main goal of this study was to use a cross-validation strategy, resample model averaging and empirical data analyses to compare seven different biometrical models for JLAM with regard to the correction for population structure and the quantitative trait loci (QTL) detection power. Three linear models and four linear mixed models with different approaches to control for population stratification were evaluated. Models A, B and C were linear models with either cofactors (Model-A), or cofactors and a population effect (Model-B), or a model in which the cofactors and the single-nucleotide polymorphism effect were modeled as nested within population (Model-C). The mixed models, D, E, F and G, included a random population effect (Model-D), or a random population effect with defined variance structure (Model-E), a kinship matrix defining the degree of relatedness among the genotypes (Model-F), or a kinship matrix and principal coordinates (Model-G). The tested models were conceptually different and were also found to differ in terms of power to detect QTL. Model-B with the cofactors and a population effect, effectively controlled population structure and possessed a high predictive power. The varying allele substitution effects in different populations suggest as a promising strategy for JLAM to use Model-B for the detection of QTL and then to estimate their effects by applying Model-C.

Molecular marker assisted broadening of the Central European heterotic groups in rye with Eastern European germplasm.

Broadening the genetic base of heterotic pools is a key to ensure continued genetic gains in hybrid breeding and extend hybrid cultivation to new areas. In the present study, two Central European heterotic pools (Carsten and Petkus) and five Eastern European open-pollinated varieties (OPVs, Pop-1 to Pop-5) were studied with the objectives to (1) investigate the genetic diversity in OPVs and the heterotic pools using molecular and field data, (2) evaluate the molecular diversity among OPVs, (3) examine the combining ability for grain yield of the OPVs when crossed with testers in field trials, and (4) develop a strategy for targeted introgression of OPV germplasm into the heterotic pools. In total, 610 S(0) plants, 347 from OPVs and 263 from heterotic pools, were developed. Clones of the S(0) plants of OPVs were crossed with two testers belonging to each heterotic pool, while clones of heterotic pools were crossed with only the opposite tester. Testcrosses were evaluated for grain yield in multi-location trials. In addition, 589 S(0) plants were fingerprinted with 30 SSR markers. The data revealed that the Carsten pool has a narrow genetic base and should be the primary target for broadening the established heterotic pattern. Mean and genetic variance suggested that Pop-2 and Pop-4 are good candidates for introgression in Petkus pool and Pop-5 in Carsten pool. Nevertheless, introgression of Pop-5 in Carsten could reduce the genetic diversity between heterotic pools. Therefore, we suggest that either selected plants of Pop-5 should be introgressed or more Eastern European germplasm should be fingerprinted and field evaluated to identify promising germplasm for broadening the established heterotic pattern.