Landessaatzuchtanstalt

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Genetic dissection of phosphorus use efficiency in a maize association population under two P levels in the field
(2021) Li, Dongdong; Wang, Haoying; Wang, Meng; Li, Guoliang; Chen, Zhe; Leiser, Willmar L.; Weiß, Thea Mi; Lu, Xiaohuan; Wang, Ming; Chen, Shaojiang; Chen, Fanjun; Yuan, Lixing; Würschum, Tobias; Liu, Wenxin
Quantitative-genetic evaluation of resistances to five fungal diseases in a large triticale diversity panel (×Triticosecale)
(2022) Miedaner, Thomas; Flath, Kerstin; Starck, Norbert; Weißmann, Sigrid; Maurer, Hans Peter
The man-made cereal triticale was fully resistant to the biotrophic diseases powdery mildew, leaf rust, yellow rust, and stem rust from its introduction in Europe in the mid-1970s until about 1990. In the following years, new races that were able to infect at least some triticale genotypes developed in all four pathogen populations, and resistance breeding came into focus. Here, we analyzed 656 winter triticale cultivars from 12 countries for resistance to these biotrophic diseases and Fusarium head blight (FHB) at up to 8 location-year combinations (environments). FHB ratings were corrected for plant height and heading stage by comparing three statistical methods. Significant (p < 0.001) genetic variances were found for all resistances with moderate to high entry-mean heritabilities. All traits showed a normal distribution, with the exception of stem rust, where the ratings were skewed towards resistance. There were no substantial correlations among the five disease resistances (r = −0.04 to 0.26). However, several genotypes were detected with multi-disease resistance with a disease rating below average for all five diseases simultaneously. In future, such genotypes must be selected primarily to cope with future challenges of less pesticide use and global climate change.
Can we abandon phosphorus starter fertilizer in maize? Results from a diverse panel of elite and doubled haploid landrace lines of maize (Zea mays L.)
(2022) Roller, Sandra; Weiß, Thea M.; Li, Dongdong; Liu, Wenxin; Schipprack, Wolfgang; Melchinger, Albrecht E.; Hahn, Volker; Leiser, Willmar L.; Würschum, Tobias
The importance of phosphorus (P) in agriculture contrasts with the negative environmental impact and the limited resources worldwide. Reducing P fertilizer application by utilizing more efficient genotypes is a promising way to address these issues. To approach this, a large panel of maize (Zea mays L.) comprising each 100 Flint and Dent elite lines and 199 doubled haploid lines from six landraces was assessed in multi-environment field trials with and without the application of P starter fertilizer. The treatment comparison showed that omitting the starter fertilizer can significantly affect traits in early plant development but had no effect on grain yield. Young maize plants provided with additional P showed an increased biomass, faster growth and superior vigor, which, however, was only the case under environmental conditions considered stressful for maize cultivation. Importantly, though the genotype-by-treatment interaction variance was comparably small, there is genotypic variation for this response that can be utilized in breeding. The comparison of elite and doubled haploid landrace lines revealed a superior agronomic performance of elite material but also potentially valuable variation for early traits in the landrace doubled haploid lines. In conclusion, our results illustrate that breeding for P efficient maize cultivars is possible towards a reduction of P fertilizer in a more sustainable agriculture.
Do we need to breed for regional adaptation in soybean? - Evaluation of genotype-by-location interaction and trait stability of soybean in Germany
(2023) Döttinger, Cleo A.; Hahn, Volker; Leiser, Willmar L.; Würschum, Tobias
Soybean is a crop in high demand, in particular as a crucial source of plant protein. As a short-day plant, soybean is sensitive to the latitude of the growing site. Consequently, varieties that are well adapted to higher latitudes are required to expand the cultivation. In this study, we employed 50 soybean genotypes to perform a multi-location trial at seven locations across Germany in 2021. Two environmental target regions were determined following the latitude of the locations. Adaptation and trait stability of seed yield and protein content across all locations were evaluated using Genotype plus Genotype-by-Environment (GGE) biplots and Shukla’s stability variance. We found a moderate level of crossing-over type genotype-by-location interaction across all locations. Within the environmental target regions, the genotype-by-location interaction could be minimised. Despite the positive correlation (R = 0.59) of seed yield between the environmental target regions and the same best-performing genotype, the genotype rankings differed in part substantially. In conclusion, we found that soybean can be grown at a wide range of latitudes across Germany. However, the performance of genotypes differed between the northern and southern locations, with an 18.8% higher mean yield in the south. This in combination with the observed rank changes of high-performing genotypes between both environmental target regions suggests that selection targeted towards environments in northern Germany could improve soybean breeding for those higher latitude regions.
The antioxidant potential of various wheat crusts correlates with AGE content independently of acrylamide
(2023) Wächter, Kristin; Longin, Carl Friedrich H.; Winterhalter, Patrick R.; Bertsche, Ute; Szabó, Gábor; Simm, Andreas
Epidemiological studies have indicated that the consumption of whole-grain products is associated with a reduced risk of cardiovascular diseases, type II diabetes, and cancer. In the case of bread, high amounts of antioxidants and advanced glycation end products (AGEs) are formed during baking by the Maillard reaction in the bread crust; however, the formation of potentially harmful compounds such as acrylamide also occurs. This study investigated the antioxidant responses of different soluble extracts from whole-grain wheat bread crust extracts (WBCEs) in the context of the asparagine, AGE, and acrylamide content. For that, we analyzed nine bread wheat cultivars grown at three different locations in Germany (Hohenheim, Eckartsweier, and Oberer Lindenhof). We determined the asparagine content in the flour of the 27 wheat cultivars and the acrylamide content in the crust, and measured the antioxidant potential using the induced expression of the antioxidant genes GCLM and HMOX1 in HeLa cells. Our study uncovered, for the first time, that the wheat crust’s antioxidant potential correlates with the AGE content, but not with the acrylamide content. Mass spectrometric analyses of WBCEs for identifying AGE-modified proteins relevant to the antioxidant potential were unsuccessful. However, we did identify the wheat cultivars with a high antioxidant potential while forming less acrylamide, such as Glaucus and Lear. Our findings indicate that the security of BCEs with antioxidative and cardioprotective potential can be improved by choosing the right wheat variety.
Generation of high oleic acid sunflower lines using gamma radiation mutagenesis and high-throughput fatty acid profiling
(2023) Rozhon, Wilfried; Ramirez, Veronica E.; Wieckhorst, Silke; Hahn, Volker; Poppenberger, Brigitte
Sunflower (Helianthus annuus L.) is the second most important oil seed crop in Europe. The seeds are used as confection seeds and, more importantly, to generate an edible vegetable oil, which in normal varieties is rich in the polyunsaturated fatty acid linoleic acid. Linoleic acid is biosynthesized from oleic acid through activity of the oleate desaturase FATTY ACID DESATURASE 2 (FAD2), which in seeds is encoded by FAD2-1, a gene that’s present in single copy in sunflowers. Defective FAD2-1 expression enriches oleic acid, yielding the high oleic (HO) acid trait, which is of great interest in oil seed crops, since HO oil bears benefits for both food and non-food applications. Chemical mutagenesis has previously been used to generate sunflower mutants with reduced FAD2-1 expression and here it was aimed to produce further genetic material in which FAD2-1 activity is lost and the HO trait is stably expressed. For this purpose, a sunflower mutant population was created using gamma irradiation and screened for fad2-1 mutants with a newly developed HPLC-based fatty-acid profiling system that’s suitable for high-throughput analyses. With this approach fad2-1 knock-out mutants could be isolated, which stably hyper-accumulate oleic acid in concentrations of 85-90% of the total fatty acid pool. The genetic nature of these new sunflower lines was characterized and will facilitate marker development, for the rapid introgression of the trait into elite sunflower breeding material.
The importance of Fusarium head blight resistance in the cereal breeding industry: Case studies from Germany and Austria
(2023) Miedaner, Thomas; Flamm, Clemens; Oberforster, Michael
Fusarium head blight (FHB) resistance in wheat and triticale has a high priority in the European Union because of the strict guidelines for the major mycotoxins deoxynivalenol (DON) and zearalenone (ZON) and the admission policy of the regulatory authorities. Potentially 70% of the arable land in Germany and about 60% in Austria can be affected by Fusarium. Although epidemics occur only in some years and/or some regions, DON and ZON are detected every year in varying amounts in wheat, rye and maize. Despite a high significance of FHB resistance in breeding companies, as validated by a recent survey, breeding progress in wheat is basically absent for FHB resistance in both countries. The main reasons are the complex inheritance of FHB resistance and the high proportions of the dwarfing allele Rht‐D1b in high‐yielding varieties promoting susceptibility. Despite this, some varieties with high FHB resistance (score 2–3 on the 1–9 scale) have been released that account, however, only for 11% and 18% of the multiplication area in Germany and Austria, respectively. For triticale, an official testing system for FHB resistance in terms of DON content exists in Germany and Austria, but not for the other cereals. Susceptibility to maize ear rot has been described in Austria, but not in Germany. Additionally, a testing system for stalk rot resistance in both countries should be established.
Spectroscopy‐based prediction of 73 wheat quality parameters and insights for practical applications
(2023) Nagel‐Held, Johannes; El Hassouni, Khaoula; Longin, Friedrich; Hitzmann, Bernd
Background and Objectives: Quality assessment of bread wheat is time-consuming and requires the determination of many complex characteristics. Because of its simplicity, protein content prediction using near-infrared spectroscopy (NIRS) serves as the primary quality attribute in wheat trade. To enable the prediction of more complex traits, information from Raman and fluorescence spectra is added to the NIR spectra of whole grain and extracted flour. Model robustness is assessed by predictions across cultivars, locations, and years. The prediction error is corrected for the measurement error of the reference methods. Findings: Successful prediction, robustness testing, and measurement error correction were achieved for several parameters. Predicting loaf volume yielded a corrected prediction error RMSECV of 27.5 mL/100 g flour and an R² of 0.86. However, model robustness was limited due to data distribution, environmental factors, and temporal influences. Conclusions: The proposed method was proven to be suitable for applications in the wheat value chain. Furthermore, the study provides valuable insights for practical implementations. Significance and Novelty With up to 1200 wheat samples, this is the largest study on predicting complex characteristics comprising agronomic traits; dough rheological parameters measured by Extensograph, micro-doughLAB, and GlutoPeak; baking trial parameters like loaf volume; and specific ingredients, such as grain protein content, sugars, and minerals.
Improving host resistance to Fusarium head blight in wheat (Triticum aestivum L.) and Gibberella ear rot in maize (Zea mays L.)
(2023) Akohoue, Félicien; Miedaner, Thomas
Fusarium head blight (FHB) in wheat and Fusarium (FER) and Gibberella ear rot (GER) in maize are major cereal diseases which reduce yield and contaminate kernels with several mycotoxins. In Europe, these diseases contribute to significant yield gaps and high mycotoxin risks across countries. However, existing management strategies related to agronomic practices are not fully effective, with some of them being cost-prohibitive. Enhancing host plant resistance is additionally required for managing the diseases more effectively and sustainably. Unfortunately, breeding for FHB resistance is challenged by complex interactions with morphological traits and the quantitative nature of the trait. In maize, available genetic resources have not been fully exploited to improve GER resistance in elite materials. In this work, we elucidated the complex interactions between FHB resistance and morphological traits, like plant height (PH) and anther retention (AR) in wheat. The effect of reduced height (Rht) gene Rht24 on AR and the contribution of genomic background (GB) to FHB resistance in semi-dwarf genotypes were also assessed. GB refers to all genomic loci, except major Rht genes, that affect the traits. To achieve this, 401 winter wheat cultivars were evaluated across five environments (location × year combination). All cultivars were genotyped using Illumina 25 K Infinium single-nucleotide polymorphism array. We performed correlation and path coefficient analysis, and combined single and multi-trait genome-wide association studies (GWAS). Our findings revealed significant genotypic correlations and path effects between FHB severity with PH and AR, which were controlled by several pleiotropic loci. FHB severity and PH shared both negatively and positively acting pleiotropic loci, while only positively acting pleiotropic loci were detected between FHB severity and AR. Rht-D1 is a major pleiotropic gene which exerted a negative effect on FHB resistance. These pleiotropic loci contribute to our understanding of the complex genetic basis of FHB resistance, and their exploitation can help to simultaneously select for FHB resistance with PH and AR. Contrary to Rht-D1b, Rht24b had no negative effect on FHB resistance and AR. This exhibits Rht24 as an important FHB-neutral Rht gene which can be integrated into breeding programs. Genomic estimated breeding values (GEBV) were calculated for each cultivar to assess GB. We observed highly negative GEBV for FHB severity within resistant wheat cultivars. Susceptible cultivars exhibited positive GEBV. Genomic prediction has a great potential and can be exploited by selecting for semi-dwarf winter wheat genotypes with higher FHB resistance due to their genomic background resistance. To tackle maize ear rot diseases, refined and stable quantitative trait loci (QTL) harboring candidate genes conferring resistances to FER and GER were identified. The effectiveness of introgression of two European flint landraces, namely “Kemater Gelb Landmais” (KE) and “Petkuser Ferdinand Rot” (PE) was evaluated. The prediction accuracy of using line performance as a predictor of hybrid performance for GER resistance was also evaluated within the two landraces. We applied a meta-QTL (MQTL) analysis based on 15 diverse SNP-based QTL mapping studies and performed gene expression analysis using published RNA-seq data on GER resistance. In total, 40 MQTL were identified, of which 14 most refined MQTL harbored promising candidate genes for use in breeding programs for improving FER and GER resistances. 28 MQTL were common to both FER and GER, with most of them being shared between silk (channel) and kernel resistances. This highlights the co-inheritance of FER and GER resistances as well as types of active resistance. Resistance genes can be transferred into elite cultivars by integrating refined MQTL into genomics-assisted breeding strategies. Afterwards, four GER resistant doubled haploid (DH) lines from both KE and PE landraces were crossed with two susceptible elite lines to generate six bi-parental populations with a total of 534 DH lines which were evaluated for GER resistance. GER severity within the six landrace-derived populations were reduced by 39−61% compared to the susceptible elite lines. Moderate to high genetic advance was observed within each population, and the use of KE landrace as a donor was generally more effective than PE landrace. This shows promise in enhancing resistance to GER in elite materials using the European flint landraces as donors. Furthermore, per se performance of 76 DH lines from both landraces was used to predict GER resistance of their corresponding testcrosses (TC). Moderate phenotypic and genomic prediction accuracy between TC and line per se performance was found for GER resistance. This implies that pre-selecting lines for GER resistance is feasible; however, TC should be additionally tested on a later selection stage to aim for GER-resistant hybrid cultivars.
Deciphering the potential of large-scale proteomics to improve product quality and nutritional value in different wheat species
(2022) Afzal, Muhammad; Longin, Friedrich
Wheat (Triticum aestivum) is one of the most important staple crops globally, which provides on average ~20% of the dietary intake of protein, starch and further important ingredients like fiber, minerals, vitamins, and essential amino acids for humans. Besides common wheat, there exist further wheat species with global to only local importance, i.e., durum, spelt, emmer and einkorn. Common wheat and durum are relatively widely cultivated whereas the other three species are cultivated only in specific regions. Apart from other functions, wheat proteins largely influence the end-use quality of products such as bread and pasta quality. Furthermore, wheat proteins can induce inflammatory reactions in humans such as celiac disease, wheat allergy and non-celiac wheat sensitivity. Thus, proteome profiles of different wheat species and cultivars within these species are of high relevance for stakeholders along the wheat supply chain. Proteomic technology has made breakthrough advancements in the recent times capable of quantifying thousands of proteins in 1.5–2 hours. Also, the wheat reference genome has been published and extended recently. These developments are extremely helpful in studying the wheat proteome at a high resolution. However, the modern large-scale proteomics has yet neither been applied to perform comparative investigation of the proteomes of different wheat species nor to study the proteomes of different types of breads and flours nor to study its application in the context of plant breeding. Therefore, we utilized modern large-scale proteomics to fill these gaps within the framework of this PhD work. First of all, an optimized data analysis pipeline was designed to deal with big proteomics data. This was necessary to estimate a multitude of quantitative genetics parameters for each protein and perform a comparative investigation of the proteomes. Optimization included implementation of data filtering based on the quantification of a protein in a given proportion of the samples, cultivars and environments. Different tests such as test for normal distribution of each protein in the context of statistical modelling and test to check the equality of variance between groups to apply the appropriate t-test were incorporated into a semi-automated workflow. In parallel, we adjusted and improved the lab methodology to deal with hundreds of samples within a short time period. We introduced a novel hybrid liquid chromatography-mass spectrometry (LC-MS) approach that combines quantification concatamer (QconCAT) technology with short microflow LC gradients and data-independent acquisition (DIA). The proposed approach measures the proteome by label-free quantification (LFQ) while concurrently providing accurate QconCAT-based absolute quantification of the key amylase/trypsin inhibitors (ATIs). These methods were then applied to compare different wheat species based on dozens of cultivars grown at multiple locations. First, we compared common wheat and spelt and identified 3,050 proteins overall. Of total proteins, 1,555 proteins in spelt and 1,166 in common wheat were only detected in a subset of the field locations. There were 1,495 and 1,604 proteins in spelt and common wheat, respectively, which were consistently expressed across all test locations in at least one cultivar. Finally, there were 84 and 193 unique proteins for spelt and common wheat, respectively, as well as 396 joint proteins, which were significantly differentially expressed between the two species. Using potentially allergenic proteins – annotated as amylase/trypsin inhibitors, serpins, and wheat germ agglutinin – we calculated an equally weighted “allergen index” that largely varied across cultivars ranging from –13.32 to 10.88 indicating the potential to select for cultivars with favorable proteome profiles. Next, we examined the proteomes of six different flours (wholegrain and superfine flours) and 14 different bread types (yeast and sourdough fermented breads and common wheat breads plus/minus bread improver) from common wheat, spelt and rye. Proteins that could cause allergies were functionally classified and comparatively measured by LFQ in flours and breads. Our findings showed that allergenic proteins were more prevalent in common wheat and spelt than rye and were not specifically degraded during bread manufacturing. In terms of abundance of the allergenic proteins, there was almost no difference between spelt and common wheat and the type of grain is likely more important for allergenicity than milling or traditional fermentation techniques. In a further study, we generated the flour reference proteomes for five wheat species, identifying at least 2,540 proteins in each species. More than 50% of the proteins significantly differed between species. Particularly, einkorn expressed 5.4 and 7.2 times less allergens and amylase/trypsin inhibitors than common wheat, respectively, emerging as a potential alternative cereal crop for people with sensitivities to cereal allergens. Lastly, we studied the application of large-scale proteomics for plant breeding. We found a significant impact of the environmental factors on protein expression. Only a fraction of proteins was stably expressed in all environments in at least one cultivar. Environmental influence was observed not only in the form of absolute expression or suppression of a certain protein at one or more environments but also in the form of low heritability (H2). High coefficients of variation across wheat cultivars indicate that the protein profiles of different cultivars vary considerably. Although, heritability was low for many proteins, we were able to identify hundreds of proteins with H²>0.5 – including key proteins for baking quality and human health. It should be possible to specifically manipulate the expression of functionally important proteins with high heritability by selecting and breeding for superior wheat cultivars along the wheat supply chain. Nevertheless, a successful implementation in plant breeding programs needs an improvement in the speed of protein quantification methods and in the validation of protein functions and annotations. In a nutshell, high number of proteins can be quantified in cereal grains utilizing cutting-edge proteomics techniques, opening new avenues for their use in the wheat supply chain. We generated lists of intriguing candidate proteins for further investigations on wheat sensitivity, and proteins with high heritability and important biological functions. Current research work has significant implications for the scientific and business communities across multiple disciplines including breeding, agriculture, cereal technology, nutritional science, health, and medicine. Political decision-makers and stakeholders in the food supply chain can benefit from the findings of this PhD project.
Mapping stem rust and leaf rust resistances in winter rye (Secale cereale L.)
(2023) Gruner, Paul; Miedaner, Thomas
Rye (Seale cereale L.) is one of the few cross-pollinating small-grain cereals and is mainly used for bread baking, biogas production and as animal feed. In its largest cultivation area (Northern, Central and Eastern Europe, including the Russian Federation) two major rust diseases, stem rust (SR) caused by Puccinia graminis f. sp. secalis and leaf rust (LR) caused by Puccinia recondita, can cause severe yield losses. Whereas LR can be found in most rye growing areas every year, SR is occurring less regularly, but can become epidemic in some years. The general occurrence of stem rust in Germany is becoming more regular, especially when hot summers provide optimum conditions for the growth and the spread of this fungus. Resistant cultivars can be a successful way to control both diseases, but SR is not assessed in the (German) variety registration and still several cultivars can be found that are susceptible or medium resistant for LR. Before the studies of this thesis were conducted, no marker-associated SR resistance gene locus was known and only six LR resistance loci had been reported. Rust resistances can be classified into all-stage resistances (ASR), that are usually caused by single R-genes and adult-plant resistances (APR), that are characterized by smaller (quantitative) effects and can only be observed in the adult-plant stage and thus make field tests mandatory. This thesis aimed on identifying resistant genotypes and respective resistance loci for SR and LR resistances in the rye genome. Two different material groups were used: biparental populations composed of inbred lines and populations composed of self-incompatible single plants. In total ten biparental populations and two additional testcross populations were studied, each constituting 68-90 genotypes. Self-incompatible populations were genetic resources from the Russian Federation, Austria and the United States of America and had 68-74 single plants each. Inbred lines were assessed in multi-environmental field trials (4-6 environments per population) and to guarantee high disease pressure, SR was artificially inoculated in contrast to naturally occurring LR in all environments. In addition, two different kind of seedling tests, one based on inoculations of entire seedling plants and one based on inoculation of detached leaves, were used to assess SR resistance. Mixed linear models were used to analyze the phenotypic data from field experiments and (mixed) cumulative logit models were used to analyze ordinal data resulting from seedling tests. Due to small sample size of a single detached leaf per genotype and isolate in self-incompatible populations, the results based on cumulative modes were cross checked with a non-parametric test. Both, progenies from biparental populations and single plants from self-incompatible populations were genotyped with single nucleotide polymorphism (SNP) based markers (Illumina iSelect 10K SNP chip or DArTseqTM) and appropriate statistical tests for phenotype-marker association were applied. This was achieved by extending phenotypic models with additive and dominant marker effects and their respective interaction with the environment or the isolates. Two marker-associated SR ASR loci (Pgs1, Pgs3.1) could be identified in biparental populations that were responsible for (large) qualitative differences between resistant and susceptible plants in the field and/or seedling stage. Additionally, 14 quantitative trait loci (QTLs) were shown to be responsible for SR APR. For LR, except one QTL found at similar position compared to a previous study, two new genes (Pr7, Pr8) and three QTLs were identified. Self-incompatible rye populations were used for the first time for association mapping and three SR resistance loci (Pgs1 - Pgs3) could be identified. Two thereof were also found within biparental mapping populations by means of QTL mapping and this was considered as prove of this new method. Throughout all studies, the natural cross-pollinating character of rye had to be considered in choosing appropriate methods and for developing rust resistant rye hybrids. This thesis includes breeding material from the largest European rye breeding companies and experiments were conducted in close cooperation with them. The characterization of breeding material for SR and LR infection, development of (new) mapping approaches, detection of resistance loci and marker candidates in the rye genome and finally the discussion of selection strategies provides a solid basis for breeders to develop the most durable SR and LR resistant rye cultivars. For scientists, new research topics could be, for example, the cloning of rye genes or a more thorough understanding of pathogen dynamics to finally achieve durable resistance in future.
Phenotypic and genomics-assisted breeding of soybean for Central Europe : from environmental adaptation to tofu traits
(2022) Kurasch, Alena; Würschum, Tobias
Soybean (Glycine max Merr.) is one of the major crops in the world providing an important source of protein and oil for food and feed; however it is still a minor crop in Central Europe. Soybean cultivation can play an important role in a more sustainable agricultural system by increasing local and regional protein production in Europe. The demand for locally produced soybean products is still growing in Europe. The key for a successful establishment of soybean cultivation in Europe is adaptation of soybean varieties to the Central European growing conditions. For the latitudinal adaptation to long-day conditions in Central to Northern Europe, an adapted early flowering and maturity time is of crucial importance for a profitable cultivation. The key traits flowering and maturity are quantitatively inherited and mainly affected by photoperiod responsiveness and temperature sensitivity. The most important loci for an early flowering and maturity are E1-E4 and the various allelic combinations condition soybean flowering and maturity time and therefore strongly contribute to the wide adaptability (Jiang et al., 2014; Tsubokura et al., 2014; M. Xu et al., 2013). Besides the main usage as protein source for animal feeding, soybean is also a very valuable source for human consumption. Tofu is enjoying ever greater popularity in Europe, as it is one of the best sources of plant protein with additional health benefits, rich in essential amino acids, beneficial lipids, vitamins, and minerals, as well as other bioactive compounds, such as isoflavones, soyasaponin, and others, (Lima et al., 2017; Zhang et al., 2018). Thus, plant breeding has to provide not only well-adapted varieties with good agronomic and quality properties, but also provide varieties well-suited to the further processing into soymilk and tofu. Therefore, a good knowledge about the breeding target, how to assess it and how it is inherited is crucial. The conducted studies covered a broad range of aspects relevant to improve a soybean breeding program. By combining environmental analysis, E-gene analysis, genomic approaches (QTL mapping and genomic prediction), and tofu phenotyping, breeder decisions become more accurate and targeted in the way of selection thereby increasing the genetic gain. In addition, combining the results of the different aspects helps to optimize the resources of a breeding program. Increasing the knowledge about the different aspects from environment to tofu QTL enables a breeder to be more precise and focused. But the more targeted and specific, the more complex a breeding program gets, which requires adequate tools to handle all the different information in a meaningful and efficient way to enable a quick and precise breeding decision.
Molecular and phenotypic diversity in populations of Fusarium culmorum on cereal hosts
(2022) Castiblanco Vargas, Eveline Valheria; Miedaner, Thomas
Fusarium head blight is one of the most devastating diseases of cereals globally and responsible for large harvest losses, not only due to the reduction in productivity but also due to the contamination of the grain with mycotoxins. The major causal agent worldwide is Fusarium graminearum; in Europe also other Fusarium species, among them Fusarium culmorum (FC) play an important role. The interaction between Fusarium species and cereals has been categorized as quantitative according to previous phenotypic and genetic observations. We studied the molecular and phenotypic diversity of natural populations of FC and how they interact with four cereals (bread wheat, durum wheat, triticale, rye) as host. Specifically, we sought (i) to understand the interaction between host and isolate, and between isolate and environment using the variance partition approach offered by mixed models applied to analyze multi-environmental studies; (ii) to identify or validate the association of Fusarium genes previously assigned as candidates using field aggressiveness and deoxynivalenol (DON) production; and (iii) to compare the application and results of the candidate gene association mapping approach applied to the same population of FC isolates but with different phenotypic data obtained from inoculation in different hosts-bread wheat and rye. Phenotyping was based on multi-environmental field experiments where each plot of the host plant was artificially inoculated with spores of the respective isolate in accordance with the experimental design. Aggressiveness was visually quantified as the percentage of spikelets with symptoms per plot and was repeatedly evaluated over time. The content of the mycotoxin deoxynivalenol (DON) in the harvested grain was evaluated by double enzyme linked immunosorbent assays (ELISA). Genes previously reported in the literature as related to aggressiveness were selected for sequencing. Using the available F. graminearum genome sequence, specific primers were constructed to amplify and sequence the most variable regions of the respective genes. The partitioning of the phenotypic variance using mixed models, for a subpopulation of 38 FC isolates in four cereal hosts, allowed to disaggregate the magnitude of the genotypic and environmental variance, and the environmental variance in turn into its different components. The genotypic variance was significant, but was exceeded by the magnitude of the environmental variance and its interactions with genotype, showing that the role of plasticity in the pathosystem of Fusarium culmorum and its cereal hosts is highly important. In contrast, the variance associated with the host factor and the interactions with host were not significant, confirmed by high values of genetic correlation amogn host. This result supports the categorization of the cereal/Fusarium culmorum interaction as unspecific and quantitatively inherited also from the view of the pathogen. For the present study, plasticity was understood as the changes in the phenotype of the pathogen that could be attributed to changes induced by the environment. Our data revealed the year as factor with the highest influence on plasticity, meaning that the isolates with high performance values under humid conditions did not exhibit the same high values under dry conditions. Because the environmental conditions are erratic between the years, the lack of a constant selection pressure in the same direction reduces the probability of achieving a speciation event per environment. The phenotypic data of the DON content in harvested grain showed a high correlation with the aggressiveness data. An association mapping study with 17 candidate genes for aggressiveness using a population of 100 isolates of FC inoculated on bread wheat revealed the significant association of the HOG1 gene, explaining 10.29% of the genetic variance of aggressiveness and 6.05% of the genetic variance corresponding to the accumulation of DON in mature grain. HOG1 is a kinase-like protein involved in the communication within the oxidative metabolism of the fungus. In a similar study using the same population of FC isolates and the same candidate genes but rye as host, the gene CUT showed a significant association with aggressiveness, explaining 16.05% of the genetic variance. The CUT gene encodes a cutinase protein, belonging to the secretome and involved in the process of unleashing the membranes and cuticles of the host plant. Taken together, our results suggest that i) field trials of breeding for resistance to FC in cereals should be carried out in several years to properly account for the genotype-by-year interaction; ii) despite the fact that molecular communication may present some type of host specificity the high plasticity guarantees that the effects on the phenotype are very similar among the cereal hosts; and iii) the high genetic correlation of aggressiveness for different cereals invites to involve non-cereal crops in the rotation plans focused on Fusarium disease management.
Analyzing resistance to ergot caused by Claviceps purpurea [Fr.] Tul. and alkaloid contamination in winter rye (Secale cereale L.)
(2022) Kodisch, Anna; Miedaner, Thomas
Ergot caused by Claviceps purpurea [Fr.] Tul. is one of the oldest well-known plant diseases leading already in medieval times to severe epidemic outbreaks. After the occurrence of honeydew, the characteristic ergot bodies called sclerotia are formed on the ear. These are containing toxic ergot alkaloids (EAs). Strict limits are set within the European Union. Rye (Secale cereale L.) as cross-pollinating crop is particularly vulnerable to ergot since the competitive situation of fungal spores and pollen during flowering. Nevertheless, even today the threat is real as agricultural practice is changing and screening studies revealed EAs in samples of the whole cereal value chain frequently. The aims were to establish a harmonized method to test ergot resistance and EA contamination in winter rye, to clarify major significant factors and their relevance and to reveal the suitability of one commercial ELISA test. Further, effort was paid to examine the covariation of ergot amount and EA content considering different factors because of prospective legislative changes. Genotypes showed significant variation for ergot severity and pollen-fertility restoration after natural infection as well as artificial inoculation whereas a high positive correlation could be found between both treatments. Additionally, variances of environment, general combining ability (GCA), specific combining ability (SCA), and interactions were significant. Although male pollen-fertility restoration was of utmost importance, the female component was also significant. This illustrates that apart from promising selection of high restoration ability the maternal restorability could be exploited in future breeding programs especially when a high pollen amount is already reached. A large-scale calibration study was performed to clarify the covariation of ergot severity, EA content (HPLC, ELISA) considering genotypes, locations, countries, years, and isolates. EA profile was rather stable across country-specific isolates although large differences regarding the EA content were detected. The moderate covariation between ergot severity and EA content (HPLC) indicates that a reliable prediction of the EA content based on ergot severity is not possible what can also not be explained by grouping effects of the factors. Further, EAs seem not to act as virulence factor in the infection process since EA content showed no relationship to disease severity. Additionally, the missing correlation of ELISA and HPLC leads to the conclusion that the ELISA is not an appropriate tool what can be used safely to screen samples regarding ergot in the daily life. The genetic variation of male-sterile CMS-single crosses was analysed in a special design without pollen in field and greenhouse to identify resistance mechanisms and to clarify whether ergot can be reduced in the female flower. At this, comparison of needle and spray inoculation revealed medium to high correlations illustrating that both methods were suitable for this research. Significant environment and genotype by environment interaction variances were detected. So, testing across several environments is necessary also without pollen. Further, small but significant genotypic variation and identification of one more ergot-resilient candidate revealed that selection of female lines could be promising to further reduce ergot. The EA content was lower for less susceptible genotypes. Thus, EA content can be considerably reduced by breeding. A strong positive correlation could be found for ergot severity and EA content when analysing 15 factorial single crosses. The male pollen-fertility restoration was also here the most relevant component but the female component contributed an obviously higher proportion for the EA content than for ergot severity. In conclusion, this thesis demonstrate that implementing of a high and environmental stable male fertility restoration ability via exotic Rf genes can effectively reduce ergot although also the female restorability enables great opportunities. The unpredictable covariation between ergot amount and EA content illustrates that both traits have to be assessed, in particular the EA content by a valid HPLC approach to guarantee food and feed safety.
Advancing soybean adaptation to Central European growth conditions with novel breeding tools
(2020) Jähne, Felix; Würschum, Tobias
According to the European Soy Monitor 2018 (European Soy Monitor, 2018), there is a wide discrepancy in the EU between market demands and general sustainability aims regarding soybean products. Europe needs to take action, if it wants to maintain its protein demands and at the same time requests a reduction in the destruction of globally important tropical and subtropical ecosystems. One step towards more sustainable soybean products lies in the increase of domestic production which has the potential to decrease soybean imports from areas of unsustainable cultivation. An augmented EU production of soybeans can be achieved for example by increasing the yield potential of soybeans in areas where successful cultivation already takes place or by expanding the cultivation area to more northern parts of Central Europe. Breeding for new, improved and adapted soybean cultivars that meet those terms, is a key activity towards that aim. This dissertation elucidates three different ways how the adaptation of soybeans to the climatic and photoperiodic conditions of Central Europe can be assisted and even accelerated: 1) By using off-season climate-controlled LED chambers to enable a speed breeding single seed descent approach. A 10 h light regime, rich in blue and deprived of far-red light emission is capable to significantly reduce and synchronise the generation time of soybeans. It was possible to shorten the life cycle for a panel of 8 soybean cultivars from different maturity groups to 77 days. This allows several generations of soybeans to be grown within one year. For the short day crops rice and amaranth on the other hand, different light quality parameters were favoured. In those crops mean flowering time was accelerated when far-red light was included in the light protocol. This underlines the importance of a crop-specific light regime in order to realise the full potential of LED-based speed breeding single seed descent. 2) By including experiments in climate-control chambers in combination with molecular tools (i.e. genomic prediction) to advance cold tolerance in soybeans. This quantitatively inherited key trait is necessary to adapt soybeans to colder regions and consequently extend growing areas of this crop to higher latitudes in Europe. In the biparental soybean population Merlin × Sigalia (103 recombinant inbred lines) three QTL for cold tolerance during pod onset were found on chromosomes 7, 11 and 13. The relatively small proportion of genotypic variance for this trait explained by these QTL underlines the quantitative nature of cold tolerance. Genomic prediction was shown to be a promising approach to select for cold stress tolerance. Scenarios with different test set sizes and prediction models were evaluated. In scenarios with smaller test set sizes prediction accuracies increased if known and confirmed QTL were included in the prediction model. 3) By incorporating citizen science into the breeding process. The citizen science project ‘1000 Gärten’ from 2016 approached this topic. Phenotypic data from soybean cultivars and breeding lines were collected by citizen scientists in 2492 gardens throughout Germany which generated a unique dataset. Among many other results this study was able to show that in 2016 and within the early maturity segment of soybeans the factor temperature influenced flowering and maturity to a higher degree than photoperiod although day length differed by over an hour between the north and the south of Germany during the time of flowering. It was shown that this admittedly challenging tool can realise a significant impact not only regarding the possibility of a highly multi-environmental screening of breeding material but also by connecting plant breeding, agriculture and potential future costumers in order to raise awareness and acceptance of a crop in larger parts of the society - a factor that may not be highlighted enough when a new crop is introduced to our agriculture. These approaches should not be seen as an alternative to classical plant breeding, but rather considered as valuable additional tools that can contribute to conventional breeding of soybeans, as well as other crops. If applied, the presented tools may assist plant breeding to pave Europe’s way towards a greener and more sustainable future that is urgently needed.
Integration of hyperspectral, genomic, and agronomic data for early prediction of biomass yield in hybrid rye (Secale cereale L.)
(2021) Galán, Rodrigo José; Miedaner, Thomas
Currently, the combination of a growing bioenergy demand and the need to diversify the dominant cultivation of energy maize opens a highly attractive scenario for alternative biomass crops. Rye (Secale cereale L.) stands out for its vigorous growth and increased tolerance to abiotic and biotic stressors. In Germany, less than a quarter of the total harvest is used for food production. Consequently, rye arises as a source of renewables with a reduced bioenergy-food tradeoff, emerging biomass as a new breeding objective. However, rye breeding is mainly driven by grain yield while biomass is destructively evaluated in later selection stages by expensive and time-consuming methods. The overall motivation of this research was to investigate the prospects of combining hyperspectral, genomic, and agronomic data for unlocking the potential of hybrid rye as a dual-purpose crop to meet the increasing demand for renewable sources of energy affordably. A specific aim was to predict the biomass yield as precisely as possible at an early selection stage. For this, a panel of 404 elite rye lines was genotyped and evaluated as testcrosses for grain yield and a subset of 274 genotypes additionally for biomass. Field trials were conducted at four locations in Germany in two years (eight environments). Hyperspectral fingerprints consisted of 400 discrete narrow bands (from 410 to 993 nm) and were collected in two points of time after heading for all hybrids in each site by an uncrewed aerial vehicle. In a first study, population parameters were estimated for different agronomic traits and a total of 23 vegetation indices. Dry matter yield showed significant genetic variation and was stronger correlated with plant height (r_g=0.86) than with grain yield (r_g=0.64) and individual vegetation indices (r_g: =<|0.35|). A multiple linear regression model based on plant height, grain yield, and a subset of vegetation indices surpassed the prediction ability for dry matter yield of models based only on agronomic traits by about 6 %. In a second study, whole-spectrum data was used to indirectly estimate dry matter yield. For this, single-kernel models based on hyperspectral reflectance-derived (HBLUP) and genomic (GBLUP) relationship matrices, a multi-kernel model combining both matrices, and a bivariate model fitted also with plant height as a secondary trait, were considered. HBLUP yielded superior predictive power than the models based on vegetation indices previously tested. The phenotypic correlations between individual wavelengths and dry matter yield were generally significant (p < 0.05) but low (r_p: =< |0.29|). Across environments and training set sizes, the bivariate model yielded the highest prediction abilities (0.56 – 0.75). All models profited from larger training populations. However, if larger training sets cannot be afforded, HBLUP emerged as a promising approach given its higher prediction power on reduced calibration populations compared to the well-established GBLUP. Before its incorporation into prediction models, filtering the hyperspectral data available by the least absolute shrinkage and selection operator (Lasso) was worthwhile to deal with data dimensionally. In a third study, the effects of trait heritability, as well as genetic and environmental relatedness on the prediction ability of GBLUP and HBLUP for biomass-related traits were compared. While the prediction ability of GBLUP (0.14 - 0.28) was largely affected by genetic relatedness and trait heritability, HBLUP was significantly more accurate (0.41 - 0.61) across weakly connected datasets. In this context, dry matter yield could be better predicted (up to 20 %) by a bivariate model. Nevertheless, due to environmental variances, genomic and reflectance-enabled predictions were strongly dependant on a sufficient environmental relationship between data used for model training and validation. In summary, to affordably breed rye as a double-purpose crop to meet the increasing bioenergy demands, the early prediction of biomass across selection cycles is crucial. Hyperspectral imaging has proven to be a suitable tool to select high-yielding biomass genotypes across weakly linked populations. Due to the synergetic effect of combining hyperspectral, genomic, and agronomic traits, higher prediction abilities can be obtained by integrating these data sources into bivariate models.
Genetic architecture of quality traits in wheat
(2021) Rapp, Matthias; Longin, Friedrich
Quality traits in wheat are of great importance, as they are required for the production of a wide range of food products. In Europe, bread wheat (Triticum aestivum ssp. aestivum) for human consumption is primarily used in pastries. For durum wheat (Triticum turgidum ssp. durum) that is used almost exclusively for pasta production, quality traits are at least as important as in bread wheat. In Central Europe, the bread wheat subspecies spelt (Triticum aestivum ssp. spelta) is characterized by a different quality compared to bread wheat. In addition, it is produced for a niche market with a particular focus on the final product quality. The high number of demanded quality traits of a wheat variety represents a great challenge for wheat breeders. Thus, knowledge about the genetic architecture and interrelation of quality traits is of high value for wheat breeding. Due to the long list of quality traits in wheat, we focused on currently important quality traits in each of the three wheat species. In durum wheat, I was interested in traits with a high importance for durum millers and pasta producers. The protein content and the sedimentation volume are of high importance for pasta producers as they influence the firmness of cooked pasta, better known as “al dente”. A low falling number may lead to brown instead of light yellow pasta, which goes back to an increased maillard reaction during pasta production and drying. The vitreousity, representing the glassy appearance of durum grains, and the thousand kernel mass influence the semolina yield and are therefore of great interest for durum millers. In the genome-wide association mapping, I identified several putative QTL for these quality traits. For the sedimentation volume, a genomic region on chromosome 1B appeared to be important. A BLAST search against the reference genomes of emmer and bread wheat revealed the Glu-B3 gene as a likely candidate. For vitreousity, genomic regions on chromosome 7A explained a larger proportion of the genotypic variance. One of these QTL, possibly related to the Pinb-2 locus, also slightly influenced the protein content. Thus, this genomic region might be a genomic reason for the positive correlation between vitreousity and protein content. For TKM we detected a putative QTL, which explained a large proportion of the genetic variance, but could not be attributed to a known gene. Besides a good performance for quality traits, a modern durum wheat variety should be complemented by a good agronomic performance, in particular a high grain yield. This poses a great challenge for plant breeders, since grain yield and protein content are negatively correlated. With regard to simultaneously improving grain yield and protein content, the protein yield or the grain protein deviation (GPD) were proposed. We evaluated those and further selection indices for their potential to be utilized for the simultaneous improvement of grain yield and protein content. Our results indicated that a simultaneous improvement of the two traits grain yield and protein content by means of an index seems possible. However, its efficiency largely depends on the weighting of the single traits. The selection for a high GPD would mainly increase the protein content whereas a selection based on protein yield would mainly improve the grain yield. Nevertheless, a combination of different indices allows balancing this selection. Compared to the primary traits grain yield and protein content, the selection indices did not essentially differ in the complexity of their genetic architecture. In bread wheat, we focused on the acrylamide precursor asparagine. Acrylamide is formed in potentially harmful concentrations when cereals are treated with high temperatures over a long period during the processing to food products. A promising strategy to reduce the acrylamide formation would be to decrease the precursors in the raw material. The wide range of variation for asparagine content showed that variety selection might have a large influence on the occurrence of acrylamide in the final product. In addition, the moderately high heritability suggested that successful breeding for lower asparagine content is possible. This conclusion is supported by the observation of no strong negative correlations between asparagine content and a number of other important traits. The genome-wide association mapping resulted in the detection of eight putative QTL, which jointly explained 78.5% of the genetic variance. A putative QTL on chromosome 7B explained with, 18.4%, the highest proportion of the genetic variance for a single marker. For spelt wheat, we assessed a high number of quality traits but placed a special emphasis on the flavor and odor of bread produced from 30 different varieties. Interestingly, we observed a significant genetic variation for bread flavor and a heritability estimate of moderate magnitude. This suggests that even for bread flavor a successful selection appears possible. Taken together, for most traits the genome-wide association mapping resulted in the detection of a high number of putative QTL. This indicates a complex genetic architecture, typical for predominantly quantitatively inherited traits. However, few of the putative QTL explained a large proportion of the genetic variance, so that they might have the potential to be used in marker-assisted selection. In order to examine the potential of genomic selection, I performed a five-fold cross validation for the different quality traits. I could confirm previous findings that the integration of QTL information as fixed effects in the genomic prediction model increased the prediction abilities considerably. The average prediction abilities for most traits suggested a high potential for genomic selection in breeding programs. In conclusion or results form a good basis for further research but more importantly already deliver valuable knowledge that can be used as guideline to advance wheat breeding programs for improved quality.
Genomics-assisted breeding strategies for quantitative resistances to Northern corn leaf blight in maize (Zea mays L.) and Fusarium diseases in maize and in triticale (× Triticosecale Wittm.)
(2021) Galiano Carneiro, Ana Luísa; Miedaner, Thomas
Fusarium head blight (FHB) in triticale (× Triticosecale Wittm.), Gibberella ear rot (GER) and Northern corn leaf blight (NCLB) in maize (Zea mays L.) are devastating crop diseases causing yield losses and/or reducing grain quality worldwide. Resistance breeding is the most efficient and sustainable approach to reduce the damages caused by these diseases. For all three pathosystems, a quantitative inheritance based on many genes with small effects has been described in previous studies. Hence, this thesis aimed to assess the potential of genomics-assisted breeding strategies to reduce FHB, GER and NCLB in applied breeding programs. In particular, the objectives were to: (i) Dissect the genetic architecture underlying quantitative variation for FHB, GER and NCLB through different quantitative trait loci (QTL) and association mapping approaches; (ii) assess the potential of genomics-assisted selection to select superior triticale genotypes harboring FHB resistance; (iii) phenotype and characterize Brazilian resistance donors conferring resistance to GER and NCLB in multi-environment trials in Brazil and in Europe; and (iv) evaluate approaches for the introgression and integration of NCLB and GER resistances from tropical to adapted germplasm. The genome-wide association study (GWAS) conducted for FHB resistance in triticale revealed six QTL that reduced damages by 5 to 8%. The most prominent QTL identified in our study was mapped on chromosome 5B and explained 30% of the genotypic variance. To evaluate the potential of genomic selection (GS), we performed a five-fold cross-validation study. Here, weighted genomic selection increased the prediction accuracy from 0.55 to 0.78 compared to the non-weighted GS model, indicating the high potential of the weighted genomic selection approach. The successful application of GS requires large training sets to develop robust models. However, large training sets based on the target trait deoxynivalenol (DON) are usually not available. Due to the rather moderate correlation between FHB and DON, we recommend a negative selection based on genomic estimated breeding values (GEBVs) for FHB severity in early breeding stages. In the long-run, however, we encourage breeders to build and test GS calibrations for DON content in triticale. The genetic architecture of GER caused by Fusarium graminearum in maize was investigated in Brazilian tropical germplasm in multi-environment trials. We observed high genotype-by-environment interactions which requires trials in many environments for the identification of stable QTL. We identified four QTL that explained between 5 to 22% of the genotypic variance. Most of the resistance alleles identified in our study originated from the Brazilian tropical parents indicating the potential of this exotic germplasm as resistance source. The QTL located on chromosome bin 1.02 was identified both in Brazilian and in European trials, and across all six biparental populations. This QTL is likely stable, an important feature for its successful employment across different genetic backgrounds and environments. This stable QTL is a great candidate for validation and fine mapping, and subsequent introgression in European germplasm but possible negative linkage drag should be tackled. NCLB is another economically important disease in maize and the most devastating leaf disease in maize grown in Europe. Virulent races have already overcome the majority of known qualitative resistances. Therefore, a constant monitoring of S. turcica races is necessary to assist breeders on the choice of effective resistances in each target environment. We investigated the genetic architecture of NCLB in Brazilian tropical germplasm and identified 17 QTL distributed along the ten chromosomes of maize explaining 4 to 31% of the trait genotypic variance each. Most of the alleles reducing the infections originated from Brazilian germplasm and reduced NCLB between 0.3 to 2.5 scores in the 1-9 severity scale, showing the potential of Brazilian germplasm to reduce not only GER but also NCLB severity in maize. These QTL were identified across a wide range of environments comprising different S. turcica race compositions indicating race non-specific resistance and most likely stability. Indeed, QTL 7.03 and 9.03/9.04 were identified both in Brazil and in Europe being promising candidates for trait introgression. These major and stable QTL identified for GER and NCLB can be introgressed into elite germplasm by marker-assisted selection. Subsequently, an integration step is necessary to account for possible negative linkage drag. A rapid genomics-assisted breeding approach for the introgression and integration of exotic into adapted germplasm has been proposed in this thesis. Jointly, our results demonstrate the high potential of genomics-assisted breeding strategies to efficiently increase the quantitative resistance levels of NCLB in maize and Fusarium diseases in maize and in triticale. We identified favorable QTL to increase resistance levels in both crops. In addition, we successfully characterized Brazilian germplasm for GER and NCLB resistances. After validation and fine mapping, the introgression and integration of the QTL identified in this study might contribute to the release of resistant cultivars, an important pillar to cope with global food security.
Breeding for resistance to Fusarium ear diseases in maize and small-grain cereals using genomic tools
(2021) Gaikpa, David Sewordor; Miedaner, Thomas
The world’s human and livestock population is increasing and there is the need to increase quality food production to achieve the global sustainable development goal 3, zero hunger by year 2030 (United Nations, 2015). However, biotic stresses such as Fusarium ear infections pose serious threat to cereal crop production. Breeding for host plant resistance against toxigenic Fusarium spp. is a sustainable way to produce more and safer cereal crops such as maize and small-grain winter cereals. Many efforts have been made to improve maize and small-grain cereals for ear rot (ER) and Fusarium head blight (FHB) resistances, using conventional and genomic techniques. Among small-grain cereals, rye had the shortest maturity period followed by the descendant, hexaploid triticale while both wheat species had the longest maturity period. In addition, rye and triticale were more robust to Fusarium infection and deoxynivalenol accumulation, making them safer grain sources for human and animal consumption. However, a few resistant cultivars have been produced by prolonged conventional breeding efforts in durum wheat and bread wheat. High genetic variation was present within each crop species and can be exploited for resistance breeding. In this thesis, the genetic architecture of FHB resistance in rye was investigated for the first time, by means of genome-wide association study (GWAS) and genomic prediction (GP). GWAS detected 15 QTLs for Fusarium culmorum head blight severity, of which two had major effects. Both weighted and unweighted GP approaches yielded higher prediction abilities than marker-assisted selection (MAS) for FHB severity, heading stage and plant height. Genomics-assisted breeding can shorten the duration of breeding rye for FHB resistance. In the past decade, genetic mapping and omics were used to identify a multitude of QTLs and candidate genes for ear rot resistances and mycotoxin accumulation in maize. The polygenic nature of resistance traits, high genotype x environment interaction, and large-scale phenotyping remain major bottlenecks to increasing genetic gains for ear rots resistance in maize. Phenotypic and molecular analyses of DH lines originating from two European flint landraces (“Kemater Landmais Gelb”, KE, and “Petkuser Ferdinand Rot”, PE) revealed high variation for Gibberella ear rot (GER) severity and three agronomic traits viz. number of days to female flowering, plant height and proportion of kernels per cob. By employing multi-SNP GWAS method, we found four medium-effect QTLs and many small-effect (10) QTLs for GER severity in combined DH libraries (when PCs used as fixed effects), none co-localized with the QTLs detected for the three agronomic traits analyzed. However, one major QTL was detected within KE DH library for GER severity. Two prioritized SNPs detected for GER resistance were associated with 25 protein-coding genes placed in various functional categories, which further enhances scientific knowledge on the molecular mechanisms of GER resistance. Genome-based approaches seems promising for tapping GER resistance alleles from European maize landraces for applied breeding. After several cycles of backcrossing and sufficient selection for agronomic adaptation traits, the resistant lines identified in this thesis can be incorporated into existing maize breeding programs to improve immunity against F. graminearum ear infection. Breeding progress can be faster using KE landrace than PE. A successful validation of QTLs identified in this thesis can pave way for MAS in rye and marker-assisted backcrossing in maize. Effective implementation of genomic selection requires proper design of the training and validation sets, which should include part of the current breeding population.
Prospects of genomic selection for disease resistances in winter wheat (Triticum aestivum L.)
(2019) Grote, Cathérine Pauline; Miedaner, Thomas
Die Ziele dieser Arbeit waren (i) die erstmalige Evaluierung des Effekts des Zwerggens Rht24 auf FHB- und STB-Resistenzen, Wuchshöhe und Ährenschieben im Vergleich zum weit genutzten Locus Rht-D1, (ii) die Untersuchung des Potenzials der nichtadaptierten QTL Fhb1 und Fhb5 für die Entwicklung von Kurzstrohweizen, (iii) die Analyse der Vorhersagegenauigkeit von GS innerhalb und zwischen Familien durch die Anwendung der beiden Modelle RR-BLUP (ridge-regression best linear unbiased prediction) und wRR-BLUP (weighted RR-BLUP) und (iv) die Berechnung des Selektionsgewinns bzw. die Bestimmung der korrekt selektierten Top-10 %-Genotypen für FHB- und STB-Resistenzen durch GS. Die Ergebnisse dieser Studie zeigten, dass das gibberellinsäuresensitive Zwerggen Rht24 auf Chromosom 6 die Wuchshöhe um durchschnittlich 8,96 cm senkte, ohne dabei die FHB- und STB-Resistenzen oder den Zeitpunkt des Ährenschiebens ungünstig zu beeinflussen. Demgegenüber senkte das weitläufig verwendete Allel Rht-D1b die FHB-Resistenz um durchschnittlich 10,05 Prozentpunkte in einer Winterweizenpopulation bestehend aus acht biparentalen Familien, die für diese Resistenzloci segregierten. Diese Arbeit hat zusätzlich aufgezeigt, dass die Resistenzallele von Fhb1 und Fhb5 die FHB-Anfälligkeit um 6,54 bzw. 11,33 Prozentpunkte reduzierten und somit bereits allein das nicht-adaptierte Allel Fhb5b in der Lage ist, den negativen Effekt von Rht-D1b auf die FHB-Resistenz im untersuchten Material auszugleichen. Das verdeutlicht, dass die Wahl der Zwerg- und Resistenzgene in Zuchtprogrammen, in denen FHB-Resistenz ein Selektionsmerkmal ist, von entscheidender Bedeutung ist. In dieser Studie wurde des Weiteren das Potenzial der GS innerhalb und zwischen Familien untersucht. Die Vorhersagegenauigkeiten innerhalb einer Familie waren für alle Zielmerkmale höher als die zwischen Familien und unterschieden sich zwischen den einzelnen Familien und Vorhersagekonstellationen. Die stärkere Gewichtung von signifikanten Markern durch das wRR-BLUP-Modell führte zu einer Verbesserung der Vorhersagegenauigkeit im Vergleich zum weit genutzten RR-BLUP-Modell, wenn einzelne Gene, wie Rht-D1, oder Major-QTL, wie Fhb5, vorhanden waren. In dieser Studie wurden die genomisch geschätzten Zuchtwerte (GEBVs) von 2.500 ungeprüften Genotypen bestimmt, basierend auf einer partiell verwandten Trainingspopulation von 1.120 Genotypen. Die 10 % FHB- und STB-resistentesten Linien und eine zufällige Stichprobe wurden unter Berücksichtigung der Wuchshöhe genomisch selektiert und phänotypisch in einem vierortigen Feldversuch evaluiert. Für die FHB-Resistenz wurde ein Selektionserfolg von 10,62 Prozentpunkten relativ zur zufällig selektierten Populationsstichprobe ermittelt. Die GS erhöhte die STB-Resistenz allerdings nur um 2,14 Prozentpunkte. Auch die Selektion von neuen Kreuzungseltern auf der Basis von GS erscheint nicht ausreichend zuverlässig, da nur 19 % der Top-10 %-Individuen korrekt selektiert wurden. Zusammenfassend stellt die GS ein wertvolles Werkzeug dar, um den Zuchtfortschritt für die komplex vererbte FHB-Resistenz über kürzere Zyklen und größere Populationen zu unterstützen. In Kombination mit der Nutzung geeigneter Zwerggene und des nicht adaptierten QTL Fhb5 kann dadurch eine Steigerung der FHB-Resistenz im Winterweizen erzielt werden.

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