The breakthrough paradox: How focusing on one form of innovation jeopardizes the advancement of science: How focusing on one form of innovation jeopardizes the advancement of science.

Research needs a balance of risk-taking in "breakthrough projects" and gradual progress. For building a sustainable knowledge base, it is indispensable to provide support for both.

Toward combining qualitative race-specific and quantitative race-nonspecific disease resistance by genomic selection.

KEY MESSAGE: A novel genomic selection strategy offers the unique opportunity to develop qualitative race-specific resistant varieties that possess high levels of the more durable quantitative race-nonspecific resistance in their genetic background. Race-specific qualitative resistance genes (R-genes) are conferring complete resistance in many pathosystems, but are frequently overcome by new virulent pathogen races. Once the deployed R-genes are overcome, a wide variation of quantitative disease resistance (QDR) can be observed in a set of previously race-specific, i.e., completely resistant genotypes-a phenomenon known as "vertifolia effect." This race-nonspecific QDR is considered to be more durable in the long term, but provides merely a partial protection against pathogens. This simulation study aimed to detangle race-specific R-gene-mediated resistance of pending selection candidates and the QDR in their genetic background by employing different genomic selection strategies. True breeding values that reflected performance data for rust resistance in wheat were simulated, and used in a recurrent genomic selection based on several prediction models and training population designs. Using training populations that were devoid of race-specific R-genes was thereby pivotal for an efficient improvement of QDR in the long term. Marker-assisted preselection for the presence of R-genes followed by a genomic prediction for accumulating the many small to medium effect loci underlying QDR in the genetic background of race-specific resistant genotypes appeared furthermore to be a promising approach to select simultaneously for both types of resistance. The practical application of such a knowledge-driven genomic breeding strategy offers the opportunity to develop varieties with multiple layers of resistance, which have the potential to prevent intolerable crop losses under epidemic situations by displaying a high level of QDR even when race-specific R-genes have been overcome by evolving pathogen populations.

Genotyping crossing parents and family bulks can facilitate cost-efficient genomic prediction strategies in small-scale line breeding programs.

KEY MESSAGE: Genomic relationship matrices based on mid-parent and family bulk genotypes represent cost-efficient alternatives to full genomic prediction approaches with individually genotyped early generation selection candidates. The routine usage of genomic selection for improving line varieties has gained an increasing popularity in recent years. Harnessing the benefits of this approach can, however, be too costly for many small-scale breeding programs, as in most genomic breeding strategies several hundred or even thousands of lines have to be genotyped each year. The aim of this study was thus to compare a full genomic prediction strategy using individually genotyped selection candidates with genomic predictions based on genotypes obtained from pooled DNA of progeny families as well as genotypes inferred from crossing parents. A population of 722 wheat lines representing 63 families tested in more than 100 multi-environment trials during 2010-2019 was for this purpose employed to conduct an empirical study, which was supplemented by a simulation with genotypic data from further 3855 lines. A similar or higher prediction ability was achieved for grain yield, protein yield, and the protein content when using mid-parent or family bulk genotypes in comparison with pedigree selection in the empirical across family prediction scenario. The difference of these methods with a full genomic prediction strategy became furthermore marginal if pre-existing phenotypic data of the selection candidates was already available. Similar observations were made in the simulation, where the usage of individually genotyped lines or family bulks was generally preferable with smaller family sizes. The proposed methods can thus be regarded as alternatives to full genomic or pedigree selection strategies, especially when pedigree information is limited like in the exchange of germplasm between breeding programs.

Simultaneous selection for grain yield and protein content in genomics-assisted wheat breeding.

KEY MESSAGE: Large genetic improvement can be achieved by simultaneous genomic selection for grain yield and protein content when combining different breeding strategies in the form of selection indices. Genomic selection has been implemented in many national and international breeding programmes in recent years. Numerous studies have shown the potential of this new breeding tool; few have, however, taken the simultaneous selection for multiple traits into account that is though common practice in breeding programmes. The simultaneous improvement in grain yield and protein content is thereby a major challenge in wheat breeding due to a severe negative trade-off. Accordingly, the potential and limits of multi-trait selection for this particular trait complex utilizing the vast phenotypic and genomic data collected in an applied wheat breeding programme were investigated in this study. Two breeding strategies based on various genomic-selection indices were compared, which (1) aimed to select high-protein genotypes with acceptable yield potential and (2) develop high-yielding varieties, while maintaining protein content. The prediction accuracy of preliminary yield trials could be strongly improved when combining phenotypic and genomic information in a genomics-assisted selection approach, which surpassed both genomics-based and classical phenotypic selection methods both for single trait predictions and in genomic index selection across years. The employed genomic selection indices mitigated furthermore the negative trade-off between grain yield and protein content leading to a substantial selection response for protein yield, i.e. total seed nitrogen content, which suggested that it is feasible to develop varieties that combine a superior yield potential with comparably high protein content, thus utilizing available nitrogen resources more efficiently.

Combining grain yield, protein content and protein quality by multi-trait genomic selection in bread wheat.

KEY MESSAGE: Simultaneous genomic selection for grain yield, protein content and dough rheological traits enables the development of resource-use efficient varieties that combine superior yield potential with comparably high end-use quality. Selecting simultaneously for grain yield and baking quality is a major challenge in wheat breeding, and several concepts like grain protein deviations have been developed for shifting the undesirable negative correlation between both traits. The protein quality is, however, not considered in these concepts, although it is an important aspect and might facilitate the selection of genotypes that use available resources more efficiently with respect to the quantity and quality of the final end products. A population of 480 lines from an applied wheat breeding programme that was phenotyped for grain yield, protein content, protein yield and dough rheological traits was thus used to assess the potential of using integrated genomic selection indices to ease selection decisions with regard to the plethora of quality traits. Additionally, the feasibility of achieving a simultaneous genetic improvement in grain yield, protein content and protein quality was investigated to develop more resource-use efficient varieties. Dough rheological traits related to either gluten strength or viscosity were combined in two separate indices, both of which showed a substantially smaller negative trade-off with grain yield than the protein content. Genomic selection indices based on regression deviations for the two latter traits were subsequently extended by the gluten strength or viscosity indices. They revealed a large merit for identifying resource-use efficient genotypes that combine both superior yield potential with comparably high end-use quality. Hence, genomic selection opens up the opportunity for multi-trait selection in early generations, which will most likely increase the efficiency when developing new and improved varieties.

Genome-wide association mapping for pre-harvest sprouting in European winter wheat detects novel resistance QTL, pleiotropic effects, and structural variation in multiple genomes.

Pre-harvest sprouting (PHS), germination of seeds before harvest, is a major problem in global wheat (Triticum aestivum L.) production, and leads to reduced bread-making quality in affected grain. Breeding for PHS resistance can prevent losses under adverse conditions. Selecting resistant lines in years lacking pre-harvest rain, requires challenging of plants in the field or in the laboratory or using genetic markers. Despite the availability of a wheat reference and pan-genome, linking markers, genes, allelic, and structural variation, a complete understanding of the mechanisms underlying various sources of PHS resistance is still lacking. Therefore, we challenged a population of European wheat varieties and breeding lines with PHS conditions and phenotyped them for PHS traits, grain quality, phenological and agronomic traits to conduct genome-wide association mapping. Furthermore, we compared these marker-trait associations to previously reported PHS loci and evaluated their usefulness for breeding. We found markers associated with PHS on all chromosomes, with strong evidence for novel quantitative trait locus/loci (QTL) on chromosome 1A and 5B. The QTL on chromosome 1A lacks pleiotropic effect, for the QTL on 5B we detected pleiotropic effects on phenology and grain quality. Multiple peaks on chromosome 4A co-located with the major resistance locus Phs-A1, for which two causal genes, TaPM19 and TaMKK3, have been proposed. Mapping markers and genes to the pan-genome and chromosomal alignments provide evidence for structural variation around this major PHS-resistance locus. Although PHS is controlled by many loci distributed across the wheat genome, Phs-A1 on chromosome 4A seems to be the most effective and widely deployed source of resistance, in European wheat varieties.

Genomic selection of parents and crosses beyond the native gene pool of a breeding program.

Genomic selection has become a valuable tool for selecting cultivar candidates in many plant breeding programs. Genomic selection of elite parents and crossing combinations with germplasm developed outside a breeding program has, however, hardly been explored until now. The aim of this study was to assess the potential of this method for commonly ranking and selecting elite germplasm developed within and beyond a given breeding program. A winter wheat (Triticum aestivum L.) population consisting of 611 in-house and 87 externally developed lines was used to compare training population compositions and statistical models for genomically predicting baking quality in this framework. Augmenting training populations with lines from other breeding programs had a larger influence on the prediction ability than adding in-house generated lines when aiming to commonly rank both germplasm sets. Exploiting preexisting information of secondary correlated traits resulted likewise in more accurate predictions both in empirical analyses and simulations. Genotyping germplasm developed beyond a given breeding program is moreover a convenient way to clarify its relationships with a breeder's own germplasm because pedigree information is oftentimes not available for this purpose. Genomic predictions can thus support a more informed diversity management, especially when integrating simply to phenotype correlated traits to partly circumvent resource reallocations for a costly phenotyping of germplasm from other programs.

Preparation of uniformly labelled 13C- and 15N-plants using customised growth chambers.

BACKGROUND: Stable isotopically labelled organisms have found wide application in life science research including plant physiology, plant stress and defense as well as metabolism related sciences. Therefore, the reproducible production of plant material enriched with stable isotopes such as 13C and 15N is of considerable interest. A high degree of enrichment (> 96 atom %) with a uniformly distributed isotope (global labelling) is accomplished by a continuous substrate supply during plant growth/cultivation. In the case of plants, 13C-labelling can be achieved by growth in 13CO2(g) atmosphere while global 15N-labelling needs 15N- containing salts in the watering/nutrient solution. Here, we present a method for the preparation of 13C and 15N-labelled plants by the use of closed growth chambers and hydroponic nutrient supply. The method is exemplified with durum wheat. RESULTS: In total, 330 g of globally 13C- and 295 g of 15N-labelled Triticum durum wheat was produced during 87 cultivation days. For this, a total of 3.88 mol of 13CO2(g) and 58 mmol of 15N were consumed. The degree of enrichment was determined by LC-HRMS and ranged between 96 and 98 atom % for 13C and 95-99 atom % for 15N, respectively. Additionally, the isotopically labelled plant extracts were successfully used for metabolome-wide internal standardisation of native T.durum plants. Application of an isotope-assisted LC-HRMS workflow enabled the detection of 652 truly wheat-derived metabolites out of which 143 contain N. CONCLUSION: A reproducible cultivation which makes use of climate chambers and hydroponics was successfully adapted to produce highly enriched, uniformly 13C- and 15N-labelled wheat. The obtained plant material is suitable to be used in all kinds of isotope-assisted research. The described technical equipment and protocol can easily be applied to other plants to produce 13C-enriched biological samples when the necessary specific adaptations e.g. temperature and light regime, as well as nutrient supply are considered. Additionally, the 15N-labelling method can also be carried out under regular glasshouse conditions without the need for customised atmosphere.

A reference-gene-based quantitative PCR method as a tool to determine Fusarium resistance in wheat.

In recent years, plant breeders made great progress in breeding Fusarium-tolerant wheat lines. However, total resistance to this genus of plant pathogenic fungi has not yet been achieved as the resistance genes are located on several distinct genetic regions. Visual scoring of disease symptoms in combination with the analysis of mycotoxins is commonly applied to assess the tolerance of new lines. Both approaches are indirect methods and do not mandatorily determine the accumulated fungal biomass. Quantitative PCR is a useful tool to assess fungal biomass based on the abundance of organism-specific DNA. The aim of this study was the development of a quantitative PCR assay for trichothecene-producing Fusarium species and to adapt this method for resistance assessment of wheat lines artificially infected with Fusarium graminearum and Fusarium culmorum. Several DNA-extraction methods for wheat samples were evaluated and optimized for downstream real-time PCR analysis and furthermore, a new reference-gene-based approach for more accurate quantification of Fusarium biomass in cereals is presented. The co-determination of a plant gene was used to compensate for unequal DNA-extraction efficiencies.

Improving and Maintaining Winter Hardiness and Frost Tolerance in Bread Wheat by Genomic Selection.

Winter hardiness is a major constraint for autumn sown crops in temperate regions, and thus an important breeding goal in the development of new winter wheat varieties. Winter hardiness is though influenced by many environmental factors rendering phenotypic selection under field conditions a difficult task due to irregular occurrence or absence of winter damage in field trials. Controlled frost tolerance tests in growth chamber experiments are, on the other hand, even with few genotypes, often costly and laborious, which makes a genomic breeding strategy for early generation selection an attractive alternative. The aims of this study were thus to compare the merit of marker-assisted selection using the major frost tolerance QTL Fr-A2 with genomic prediction for winter hardiness and frost tolerance, and to assess the potential of combining both measures with a genomic selection index using a high density marker map or a reduced set of pre-selected markers. Cross-validation within two training populations phenotyped for frost tolerance and winter hardiness underpinned the importance of Fr-A2 for frost tolerance especially when upweighting its effect in genomic prediction models, while a combined genomic selection index increased the prediction accuracy for an independent validation population in comparison to training with winter hardiness data alone. The prediction accuracy could moreover be maintained with pre-selected marker sets, which is highly relevant when employing cost reducing fingerprinting techniques such as targeted genotyping-by-sequencing. Genomic selection showed thus large potential to improve or maintain the performance of winter wheat for these difficult, costly, and laborious to phenotype traits.

Trichothecenes in Cereal Grains - An Update.

Trichothecenes are sesquiterpenoid mycotoxins produced by fungi from the order Hypocreales, including members of the Fusarium genus that infect cereal grain crops. Different trichothecene-producing Fusarium species and strains have different trichothecene chemotypes belonging to the Type A and B class. These fungi cause a disease of small grain cereals, called Fusarium head blight, and their toxins contaminate host tissues. As potent inhibitors of eukaryotic protein synthesis, trichothecenes pose a health risk to human and animal consumers of infected cereal grains. In 2009, Foroud and Eudes published a review of trichothecenes in cereal grains for human consumption. As an update to this review, the work herein provides a comprehensive and multi-disciplinary review of the Fusarium trichothecenes covering topics in chemistry and biochemistry, pathogen biology, trichothecene toxicity, molecular mechanisms of resistance or detoxification, genetics of resistance and breeding strategies to reduce their contamination of wheat and barley.

Targeted modification of plant genomes for precision crop breeding.

The development of gene targeting and gene editing techniques based on programmable site-directed nucleases (SDNs) has increased the precision of genome modification and made the outcomes more predictable and controllable. These approaches have achieved rapid advances in plant biotechnology, particularly the development of improved crop varieties. Here, we review the range of alterations which have already been implemented in plant genomes, and summarize the reported efficiencies of precise genome modification. Many crop varieties are being developed using SDN technologies and although their regulatory status in the USA is clear there is still a decision pending in the EU. DNA-free genome editing strategies are briefly discussed because they also present a unique regulatory challenge. The potential applications of genome editing in plant breeding and crop improvement are highlighted by drawing examples from the recent literature.