Browsing by Subject "QTL"

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Biometrical Analyses of Epistasis and the Relationship between Line per se and Testcross Performance of Agronomic Traits in Elite Populations of European Maize (Zea mays L.)
(2005) Mihaljevic, Renata; Melchinger, Albrecht E.
Relations of yield and other important agronomic traits of inbred lines to the same traits in hybrids have been studied from the time of initiation of hybrid breeding to the present. Because crossing lines to a tester and conducting yield trials are expensive and time-consuming, reliable information on inbred lines that is indicative of their testcross performance is crucial for optimum testing schemes in hybrid breeding as well as simultaneous improvement of commercial hybrids and their inbred parents. It has therefore been of great importance to determine the magnitude of correlation between line per se performance (LP) and testcross performance (TP) and investigate if epistasis influences this correlation. The comprehensive study on hand was performed with five populations (F3 to F6 lines) differing in size (ranging from 71 to 344), level of inbreeding, and the number of common parents. The populations employed were derived from three biparental crosses within the heterotic pool of European elite flint maize (Zea mays L.). All five populations were evaluated for TP (using an unrelated dent tester inbred) of five agronomically important quantitative traits: grain yield, grain moisture, kernel weight, protein concentration, and plant height. Four of these populations were also evaluated for LP of the same five traits. The objectives were to (i) estimate phenotypic and genotypic correlations between LP and TP within four populations for all five traits, (ii) map quantitative trait loci (QTL) for LP and TP in four and five populations, respectively, for all five traits, (iii) validate estimated QTL effects and positions for TP by assessing QTL congruency among testcross populations differing in size and genetic background, (iv) determine the value of LP-QTL for the prediction of TP, (v) estimate the importance of epistatic effects for LP and TP of grain yield and grain moisture by generation means analysis as well as genome-wide testing for epistatic marker pairs, and (vi) draw conclusions regarding the prospects of marker-assisted selection (MAS). Genotypic correlations between LP and TP, rg(LP, TP), estimated herein were comparable with those obtained for European flint or U.S. dent material. The magnitude of rg(LP, TP) was trait-specific: for traits of high heritability, i.e. grain moisture, kernel weight, protein concentration, and plant height, estimates were generally larger than 0.7 across all four populations, whereas for grain yield, estimates were consistently lower and did not exceed the intermediate level of 0.5. For grain yield, lowest rg(LP, TP) were estimated with lowest precision (largest confidence intervals). This requires testing for both LP and TP and/or combining the data in a selection index to ensure sufficient inbred performance (seed production) and yield improvement. However, combined selection for LP and TP proved less efficient than sole selection for TP unless unadapted material was employed. For kernel weight, protein concentration, and plant height, we detected "large" congruent QTL across testcross populations derived from the same cross, which individually explained up to 46% of the validated genotypic variance p. However, as the p values estimated from validation were still below the corresponding heritability estimates, MAS will be superior to phenotypic selection only if it is more cost-efficient. For the above traits, similar numbers of QTL for LP and TP were detected across populations. More than half of the QTL regions detected for LP were in common for LP and TP in the largest population (N = 280). To assess the value of QTL identified for LP in predicting TP, we calculated the genotypic correlation rg(MLP, YTP). This parameter assesses QTL congruency for LP and TP quantitatively and is thus the key parameter for assessing the prospects of MAS. The number of common QTL for LP and TP (qualitative QTL congruency) was generally not indicative of the magnitude of rg(MLP, YTP) due to the differences in the effect size of the respective QTL detected for LP and used for the prediction of TP. For all traits, rg(MLP, YTP) were smaller than rg(LP, TP). This is because rg(MLP, YTP) is only predictive for the validated proportion of genotypic variance explained by the QTL for LP, which was generally below 50% because of the limited power of QTL detection, in particular with small sample sizes below 100. Only if QTL detected for LP explain a substantial proportion of the genotypic variance, MAS based on these QTL can be applied, provided it is more cost-efficient than an indirect phenotypic selection for TP based on LP. QTL detection power was drastically reduced for the complex trait grain yield with a presumably large number of small QTL underlying its genetic architecture. Thus, the number of common QTL for LP and TP as well as the QTL congruency across testcross populations was much lower for grain yield than the other four traits. Estimated gene action of QTL detected for LP was primarily additive for grain yield. Evidence for dominance and/or epistasis, which may be a reason for the low rg(LP, TP) and the low number of common QTL for LP and TP was generally weak. Both generation means analysis for LP and TP and genome-wide search for epistatic marker pairs yielded no evidence for epistasis. This is not only because the detected epistatic effects could not be validated, but also because there is low chance to find epistasis unless the generation examined displays the full epistatic variance such as expected from doubled haploids produced from an F1 cross. Thus, it is anticipated that the relative importance of epistatic effects in hybrid maize breeding may strongly increase with the currently happening shift in line development from recurrent selfing towards the production of doubled haploids.
Dissection of the genetic architecture of stalk mechanical strength and in vivo haploid induction in maize
(2016) Hu, Haixiao; Melchinger, Albrecht E.
Stalk lodging causes yield losses in maize cultivation ranging from 5 to 20% annually worldwide and stalk mechanical strength is widely accepted as an indirect indicator for its measurement. QTL mapping can reveal the genetic basis of stalk strength and provide information about markers suitable for marker-assisted selection (MAS). Constantly increasing market demands urge maize geneticists and breeders not only to enhance the field performance of new hybrids, but also to improve the breeding process. During the last decade, advances in the double haploid (DH) technology based on in vivo haploid induction (HI) shifted the breeding paradigm and greatly accelerated the breeding process in maize. Further spread of DH technology urgently demands a simple but efficient way for developing new inducers, which could be achieved by introducing the mandatory QTL/gene(s) of HI to advanced breeding lines. Therefore, the main goal of my thesis was to dissect the genetic architecture of stalk strength and detect the mandatory genomic region(s) of HI using genome-wide molecular markers. Several methods have been developed and applied in the literature to evaluate stalk mechanical strength, among which the rind penetrometer resistance (RPR) is a simple, rapid and non-destructive measurement during data collection, whereas stalk bending strength (SBS) is more closely associated with stalk lodging in the field. According to common knowledge in the mechanics of materials, SBS is reflected by the maximum load exerted to breaking (Fmax), the breaking moment (Mmax) and the critical stress (σmax). Thus, to have a complete understanding of the genetic architecture of stalk strength in maize, RPR and SBS (measured by Fmax, Mmax and σmax) were used to characterize stalk strength in our study. Utilizing a segregating population with 216 recombinant inbred lines, our analysis showed that stalk strength traits, RPR and SBS, have high heritability, ranging from 0.75 to 0.91. Nine QTL and one epistatic interaction between QTL were detected for RPR. Two, three and two QTL were detected for Fmax, Mmax and σmax, respectively. All QTL showed minor effects and only one QTL on chromosome 10 had overlapping support intervals between RPR and SBS. Co-locations of QTL and high positive correlations between stalk strength traits and other stalk traits suggested presence of pleiotropism and a complex genetic architecture of stalk strength. Owing to lack of major QTL, MAS solely based on molecular markers was found to be less effective than classical phenotypic selection for stalk strength. However, for SBS we observed considerably higher proportions of genetic variance explained by a genomic selection approach than obtained in QTL mapping with cross validation. Therefore, genomic selection might be a promising tool to improve the efficiency of breeding for stalk strength. All QTL mapping studies conducted hitherto for unraveling the genetic architecture of HI rate detected a major QTL, termed qhir1, in bin 1.04. Dong et al. (2013) further narrowed down this QTL to a 243 kb region. Considering the complex genetic architecture of HI and genetic background noise possibly affecting fine mapping of qhir1, we attempted to validate these results with an alternative approach before embarking on map-based gene isolation. Utilizing 51 maize haploid inducers and 1,482 non-inducers collected worldwide, we were able to investigate the genetic diversity between inducers and non-inducers and detect genomic regions mandatory for HI. The genetic diversity analyses indicated that the inducer group was clearly separated from other germplasm groups and had high familial relatedness. Analyzing our data by a case-control association approach failed because the segregation of HI was heavily confounded with population structure. Moreover, selective sweep approaches commonly used in the literature that are designed for capturing selective sweeps in a long-term evolutionary context failed due to high familial relatedness among inducers. To solve this problem, we developed a novel genome scan approach to detect fixed segments among inducers. With this approach, we detected a segment, termed qhir12, 4.0 Mb in length, within the support interval of the qhir1. This segment was the longest genomic segment detected by our novel approach and was entirely absent in all non-inducers analyzed. However, qhir12 has no overlap with the fine mapping region of Dong et al. (2013), termed qhir11. This indicates that the genomic region harboring the mandatory gene of HI should be confirmed by further experiments to corroborate its existence and identify its location in the maize genome.
Effects of non-adapted quantitative trait loci (QTL) for Fusarium head blight resistance on European winter wheat and Fusarium isolates
(2010) Ohe, Christiane von der; Miedaner, Thomas
Fusarium head blight (FHB), caused by Fusarium graminearum and F. culmorum, is a devastating disease responsible for tremendous damage in wheat fields and contamination of grain with mycotoxins deoxynivalenol (DON) and nivalenol (NIV), rendering the harvest unsafe for human and animal consumption. The variability of Fusarium populations is high and changes in aggressiveness, chemotypes or species within and among Fusarium populations are known. Stable FHB resistance combined with high yield is one main target in wheat breeding programs. Mapping studies detected several quantitative trait loci (QTL) for FHB resistance in non-adapted sources, such as Sumai3 from China. The two most important and commonly used major QTL are located on chromosome 3BS (Fhb1) und 5A (Qfhs.ifa-5A). However, negative side effects of non-adapted resistance sources introgressed in elite winter wheat material are feared in Europe. Furthermore, the stability of the QTL effect against changing Fusarium populations is unknown. The objectives of this research were to analyze whether (1) the QTL Fhb1 and Qfhs.ifa-5A introgressed from a non-adapted resistance source into two winter wheat varieties have possible side effects on agronomic and quality performance, (2) 3-ADON and 15-ADON chemotypes are significantly different in their aggressiveness and DON production, (3) competition among Fusarium isolates in mixtures exists, and if so, how the resistant host will influence this competition. In conclusion, both resistance QTL are effective and stable in elite spring and winter wheat backgrounds. For improvement of FHB resistance both QTL are valuable, but Qfhs.ifa-5A would suffice for European breeding programs. Due to chemotype shifts, 3-ADON isolates could pose a greater risk to food safety than 15-ADON but breeding and use of highly resistant lines can reduce the risks associated with DON in wheat. Accordingly, resistant spring wheat lines were less affected by the tested Fusarium isolates and mixtures and, therefore, confirmed a high stability of these QTL. Directed selection of highly aggressive isolates due to the resistance QTL seems to be unlikely in the short term.
Genetic analysis of resistance to ear rot and mycotoxin contamination caused by Fusarium graminearum in European maize
(2012) Martin, Matthias; Melchinger, Albrecht E.
Maize is affected by a number of diseases. Among the various ear rots of maize, Gibberella ear rot (GER) caused by Fusarium graminearum is prevalent in Central Europe. This fungal pathogen produces secondary metabolites (mycotoxins), which adversely affect the health of humans and animals. Two important mycotoxins are the immunosuppressant deoxynivalenol (DON) and the mycoestrogen zearalenone (ZEA). The most efficient method to reduce mycotoxin contamination in maize is cultivation of resistant varieties. However, resistance breeding using classical phenotypic selection is laborious and time-consuming. Therefore, marker-assisted selection (MAS) may be a promising alternative to classical selection. Furthermore, for setting up a breeding program, knowledge about the relevance of the different modes of gene action and genotypic correlations among resistance and agronomic traits is required. The objectives of this study were to (1) estimate quantitative genetic parameters for GER severity and mycotoxin concentration in connected populations of doubled haploid (DH) lines, (2) map quantitative trait loci (QTL) for GER resistance and reduced mycotoxin contamination in these populations, (3) examine the congruency of QTL in these populations, (4) evaluate the prospects of using MAS to breed for GER resistance and reduced mycotoxin contamination, (5) estimate the genotypic correlation between the resistance of DH lines per se and the resistance of their testcrosses, (6) evaluate the influence of selection for increased resistance on agronomic performance of hybrids and (7) examine the relevance of different modes of gene action involved in the expression of the resistance in flint maize. Three field experiments were conducted, each of which comprised a different set of plant material. Experiment I comprised five DH line populations derived from the following F1 crosses among elite flint inbred lines: D152×UH006, D152×UH007, UH007×UH006, UH009×UH006 and UH009×UH007. Experiment II comprised testcross progenies of 94 DH lines and a dent single cross tester. Experiment III comprised the five F1 crosses, from which the DH populations had been derived, the F2 and the first backcross generations to the parents (BC1-P1, BC1-P2) as well as the two parent lines of each cross. Plants were artificially infected with spores of F. graminearum shortly after mid-silking using the silk channel inoculation technique. The DH lines were genotyped with simple sequence repeat (SSR) DNA markers, genetic linkage maps were constructed and QTL analyses were performed for resistance to GER, DON and ZEA contamination. Estimates of genotypic and genotype-by-environment interaction variances in Experiment I for GER severity and mycotoxin concentration were significant and heritabilities were moderately high to high in all populations. Thus, differences among DH lines for the resistance traits were mainly caused genetically and the resistance response varied depending on the environment. Owing to the effectiveness of artificial inoculation, the prospects are good to improve line resistance using a small number of test environments. QTL were detected in the four largest populations. Depending on the population, the mapped QTL together explained 21-51% of the genotypic variance for GER severity and 19-45% for DON concentration and 52% for ZEA concentration. Additive gene action was more important than digenic interactions of QTL, as indicated by the number of QTL having significant additive effects, their relative contributions to the total genotypic variance explained and the magnitude of their effects. Colocalized QTL for resistance to GER and mycotoxin contamination were identified in each mapping population. This was in agreement with strong genotypic correlations among these traits. QTL located at similar positions were detected across three populations in two chromosomal regions and across two populations in additional two regions. The results of this study indicated that a combination of classical phenotypic selection and MAS is a promising strategy for resistance breeding. In Experiment II, significant genotypic variation for resistance in lines and testcrosses showed that selection will be successful in both groups. Owing to low genotypic correlations between lines and testcrosses, however, resources should be mainly allocated to the evaluation of GER in testcrosses. Correlations of resistance with agronomic traits were weak or not significant. Therefore, selection for resistance and better agronomic performance can be carried out simultaneously. In Experiment III, generation means analysis indicated a prevalence of additive gene action for resistance. Significant dominance effects were found in only one cross for resistance to GER, but in four crosses for resistance to DON contamination. Owing to prevalence of additive gene action, the prospects are good to improve the resistance of the flint germplasm and to accumulate more favorable gene combinations in future breeding lines. Comparing the hybrid performance of flint×flint crosses of Experiment II and flint×dent crosses of Experiment III with their corresponding mid-parent performances indicated mid-parent heterosis for resistance. Therefore, prediction of hybrid performance based on performance of their parents will be possible only to a very limited extent. Future research should focus on fine mapping and validating of the detected QTL. For an efficient use of the QTL in a marker-assisted breeding program, knowledge about their effects in different genetic backgrounds is needed. Of particular importance are thereby the QTL effects in flint×dent crosses, which represent the preferred type of hybrid in Central European maize breeding programs.
Genetic approaches to dissect iron efficiency in maize
(2015) Benke, Andreas; Stich, Benjamin
Maize is susceptible to severe Fe-deficiency symptoms when growing on soils with high pH. Therefore, development of Fe-efficient maize genotypes would aid to overcome Fe limitation on these soils. However, Fe-efficiency is a quantitative trait depending on complex mechanism interactions. The determination of these mechanisms would provide a better understanding of the complex trait Fe-efficiency. In the actual study, the determination of Fe-efficiency involved mechanisms were tackled by population and quantitative genetics. In fact, population genetics facilitate the discovery of genes being important to crop improvement based on a comparison of gene evolution and its ancestral genetic material. Linkage mapping and association analyses require both phenotypic variation and polymorphic markers to determine important quantitative trait loci (QTL). The objective of this research was to dissect the genetic architecture of Fe-efficiency in maize by applying different genetic approaches. Artificial selection during domestication and (or) crop improvement can result in limitation of sequence variation at candidate genes that could limit their detection by quantitative genetic approaches. The objectives of our study were to (i) describe patterns of sequence variation of 14 candidate genes for mobilization, uptake, and transport of Fe in maize, as well as regulatory function and (ii) determine if these genes were targets of selection during domestication. This study was based on 14 candidate genes sequences of 27 diverse maize inbreds, 18 teosinte inbreds, and one Zea luxurians strain as an outgroup. The experimental results suggested that the majority of candidate genes for Fe-efficiency examined in this study were not target of artificial selection. Nevertheless, the genes NAAT1, NAS1, and MTK coding for enzymes involved in phytosiderophore production, NRAMP3 responsible for Fe remobilization during germination, and YS1 transporting PS-Fe-complexes into the root showed signatures of selection. These genes might be important for the adaptation of maize to diverse environments with different Fe availabilities. This in turn suggests, that Fe-efficiency was an adaptive trait during maize domestication from teosinte. Identification of QTL provides information on the chromosomal locations contributing to the quantitative variation of complex traits. The benefit of QTL mapping compared to mutant screenings is the possibility to detect multiple genes which may be associated with the phenotypic trait. The objectives of our studies were to (i) identify QTLs for morphological and physiological traits related to Fe homeostasis, (ii) analyze Fe-dependent expression levels of genes known to be involved in Fe homeostasis as well as positional candidate genes from QTL analysis, and (iii) identify QTLs which control the mineral nutrient concentration difference. Our studies were based on experimental data of 85 genotypes from the IBM population cultivated in a hydroponic system. The QTL mapping of morphological and physiological traits provided new putative candidate genes like Ferredoxin 1, putative ferredoxin PETF, MTP4, and MTP8 which complement the genes already known as being responsible for efficient Fe homeostasis at both, deficient and sufficient Fe regime. Furthermore, the candidate gene expression indicated a trans-acting regulation for DMAS1, NAS3, NAS1, FDH1, IDI2, IDI4, and MTK. The mineral element trait QTL confidence intervals comprised candidate genes that sequestrate Cd in vacuoles (HMA3), transport Fe2+into the root cells (ZIP10), protect the cell against oxidative stress (glutaredoxin), ensure micro nutrient homeostasis during sufficient iron regime (NRAMP2), regulate protein activities (PP2C), and prevent deleterious accumulation and interaction of specific elements within cells (PHT1;5, ZIP4). Association mapping is promising to overcome the limitations of low allele diversity and absent recombinations events causing poor resolution in detecting QTL by linkage mapping. In order to unravel the genetic architecture of Fe-efficiency a vast association mapping panel comprising 267 maize inbred lines was used to (i) detect polymorphisms affecting the morphological/physiological trait formation and (ii) fine map QTL confidence intervals determined according to linkage mapping. Some of the SNPs located beyond coding regions of genes that might be important cis-binding-sites for transcription factors. Furthermore, genes detected at the Fe-deficient regime indicate to be involved in universal stress response. However, genes linked to SNPs detected at Fe-sufficient regime might comprise alleles of Fe inefficient genotypes causing inferior trait expression. The combination of several approaches provided a valuable resource of candidate genes which might aid to increase our understanding of the mechanisms of Fe-efficiency in maize and foster the efforts in breeding superior cultivars by applying molecular marker techniques.
Genetic dissection of phosphorus use efficiency and genotype-by-environment interaction in maize
(2022) Li, Dongdong; Li, Guoliang; Wang, Haoying; Guo, Yuhang; Wang, Meng; Lu, Xiaohuan; Luo, Zhiheng; Zhu, Xintian; Weiß, Thea Mi; Roller, Sandra; Chen, Shaojiang; Yuan, Lixing; Würschum, Tobias; Liu, Wenxin
Genotype-by-environment interaction (G-by-E) is a common but potentially problematic phenomenon in plant breeding. In this study, we investigated the genotypic performance and two measures of plasticity on a phenotypic and genetic level by assessing 234 maize doubled haploid lines from six populations for 15 traits in seven macro-environments with a focus on varying soil phosphorus levels. It was found intergenic regions contributed the most to the variation of phenotypic linear plasticity. For 15 traits, 124 and 31 quantitative trait loci (QTL) were identified for genotypic performance and phenotypic plasticity, respectively. Further, some genes associated with phosphorus use efficiency, such as Zm00001eb117170, Zm00001eb258520, and Zm00001eb265410, encode small ubiquitin-like modifier E3 ligase were identified. By significantly testing the main effect and G-by-E effect, 38 main QTL and 17 interaction QTL were identified, respectively, in which MQTL38 contained the gene Zm00001eb374120, and its effect was related to phosphorus concentration in the soil, the lower the concentration, the greater the effect. Differences in the size and sign of the QTL effect in multiple environments could account for G-by-E. At last, the superiority of G-by-E in genomic selection was observed. In summary, our findings will provide theoretical guidance for breeding P-efficient and broadly adaptable varieties.
Genetic variation for cold tolerance in two nested association mapping populations
(2023) Revilla, Pedro; Butrón, Ana; Rodriguez, Víctor Manuel; Rincent, Renaud; Charcosset, Alain; Giauffret, Catherine; Melchinger, Albrecht E.; Schön, Chris-Carolin; Bauer, Eva; Altmann, Thomas; Brunel, Dominique; Moreno-González, Jesús; Campo, Laura; Ouzunova, Milena; Álvarez, Ángel; Ruíz de Galarreta, José Ignacio; Laborde, Jacques; Malvar, Rosa Ana
Cold reduces maize (Zea mays L.) production and delays sowings. Cold tolerance in maize is very limited, and breeding maize for cold tolerance is still a major challenge. Our objective was to detect QTL for cold tolerance at germination and seedling stages. We evaluated, under cold and control conditions, 919 Dent and 1009 Flint inbred lines from two nested association mapping designs consisting in 24 double-haploid populations, genotyped with 56,110 SNPs. We found a large diversity of maize cold tolerance within these NAM populations. We detected one QTL for plant weight and four for fluorescence under cold conditions, as well as one for plant weight and two for chlorophyll content under control conditions in the Dent-NAM. There were fewer significant QTL under control conditions than under cold conditions, and half of the QTL were for quantum efficiency of photosystem II. Our results supported the large genetic discrepancy between optimal and low temperatures, as the quantity and the position of the QTL were very variable between control and cold conditions. Furthermore, as we have not found alleles with significant effects on these NAM designs, further studies are needed with other experimental designs to find favorable alleles with important effects for improving cold tolerance in maize.
Genome-wide association mapping of molecular and physiological component traits in maize
(2013) Riedelsheimer, Christian; Melchinger, Albrecht E.
Genome-wide association (GWA) mapping emerged as a powerful tool to dissect complex traits in maize. Yet, most agronomic traits were found to be highly polygenic and the detected associations explained together only a small portion of the total genetic variance. Hence, the majority of genetic factors underlying many agronomically important traits are still unknown. New approaches are needed for unravelling the chain from the genes to the phenotype which is still largely unresolved for most quantitative traits in maize. Instead of further enlarging the mapping population to increase the power to detect even smaller QTL, this thesis research aims to present an alternative route by mapping not the polygenic trait of primary interest itself, but genetically correlated molecular and physiological component traits. As such components represent biological sub-processes underlying the trait of interest, they are supposed to be genetically less complex and thus, more suitable for genetic mapping. Using large diversity panels of maize inbred lines, this approach is demonstrated with (i) biomass yield by using metabolites and lipids as molecular component traits and with (ii) chilling sensitivity by using physiological component traits such as photosynthesis parameters derived from chlorophyll fluorescence measurements. In a first step, we developed a sampling and randomization scheme which allowed us to obtain metabolic and lipid profiles from large-scale field trials. Both profiles were found to be inten- sively structured reflecting their functional grouping. They also showed repeatabilities higher than in comparable profiles obtained in previous studies with the model plant Arabidopsis under controlled conditions. By applying GWAS with 56,110 SNPs to metabolites and lipids, large-scale genetic associations explaining more than 30 % of the genetic variance were detected. Confounding with structure was found to be a problem of less extent for molecular components than for agronomic traits like flowering time. The lipidome was also found to show a multilevel control architecture similar as employed in controlling complex mechanical systems. In several instances, direct links between candidate genes underlying the detected associations and agronomic traits could be established. An example is cinnamoyl-CoA reductase, a key enzyme in the lingin biosynthesis pathway. It was found to be a candidate gene underlying a major QTL found for several intermediates in the lignin biosynthesis pathways. These intemediates were in turn found to be correlated with plant height, lignin content, and dry matter yield at the end of the vegetation period. The different signs of these correlations indicated that the relationships between pathway intermediates and the final product is not simple. Directly modeling complex traits with individual component traits may therefore require consideration of feedback loops and other interdependencies. Such connections were however found difficult to be established with physiological components underlying chilling sensitivity. The main reasons for this were the weak correlations between physiological components under controlled conditions and chilling sensitivity in the field as well as high levels of genotype × environment interactions caused by the complex and environment- dependent responses of maize after perception of chilling temperatures. The approach explored in this thesis research uses component traits to gain biological insights about the genetic control of biomass yield and chilling sensitivity evaluated in diverse populations of still manageable sizes. We showed that GWAS with 56k SNPs can identify large additive effects for component traits correlated with these traits. For mapping epistatic interactions and rare variants, classical linkage mapping with biparental populations will be a reasonable complementary approach. However, controlling and modeling genotype × environment interactions remains an important issue for understanding the genetic basis of especially chilling sensitivity. If the goal is merely to predict the phenotypic value in a given set of en- vironments, black-box genomic selection methods with either SNPs, molecular profiles, or a combination of both, are very promising strategies to achieve this goal.
Gibberella ear rot resistance in European maize : genetic analysis by complementary mapping approaches and improvement with genomic selection
(2022) Han, Sen; Melchinger, Albrecht E.
During the last decades, implementation of molecular markers such as single nucleotide polymorphisms (SNPs) has transformed plant breeding practices from conventional phenotypic selection to marker-assisted selection (MAS) and genomic selection (GS) that are more precise, faster and less resource-consuming. In this dissertation, we investigated these three selection approaches for improving the polygenic trait Gibberella ear rot (GER) resistance in maize (Zea mays L.), which is an important fungal disease in Europe and North America leading to reduced grain yield and grain contaminated with mycotoxins such as deoxynivalenol (DON) and zearalenone (ZON). Three different sets of materials were evaluated in multiple environments and analyzed for different objectives. In the first study, five flint doubled-haploid (DH) families (with size 43 to 204) inter-connected at various levels through common parents, were generated in an incomplete half-diallel design with four parental lines developed by the University of Hohenheim. Significant genotypic variances and generally high heritabilities were observed for all three traits (i.e., GER, DON and days to silking (DS)) in all families, implying good prospects for resistance breeding and phenotypic selection against GER across different environments in European maize germplasm. Genetic correlations were extremely tight between DON and GER and moderately negative for DS with DON or GER, suggesting that indirect selection against GER would be efficient to reduce DON, but maturity should be considered in GER resistance breeding. Using a high-density consensus map with 2,472 marker loci, we compared classical bi-parental mapping of QTL (quantitative trait locus/loci) with multi-parental QTL mapping conducted with joint families and using four different biometric models. Multi-parental QTL mapping models identified all and even further QTL than the bi-parental QTL mapping model conducted within each family. Interestingly, QTL for DON and GER were mostly family-specific, yet multiple families had several common QTL for DS. Many QTL displayed large additive effects and most favorable alleles originated from the highly resistant parent. Interactions between detected QTL and genetic background (family) were rare and had comparatively small effects. Multi-parental QTL mapping models generally did not yield higher prediction accuracy than the bi-parental QTL mapping model for all traits. In the second study, two diversity panels consisting of 130 elite European dent and 114 flint lines, respectively, from the University of Hohenheim were evaluated and subject to a genome-wide association study within each pool. Similar to the first study, highly significant genotypic and genotype × environment interaction variances were observed for GER, DON and DS. Heritabilities were moderately high for GER and DON and high for DS in both pools. Estimated genomic correlations between pools were close to zero for DON and DS, and slightly higher for GER. The detected QTL for DON were all specific to each heterotic pool and none of them was in common with previously detected QTL. Furthermore, no QTL was detected for GER and DS in both pools. Genomic prediction (GP) across pools yielded low or even negative prediction accuracy for all traits. When the training set (TS) size was increased by combining lines from both heterotic pools, the combined-pool GP approaches had no higher prediction accuracy than the within-pool GP approach. Different from expectation, method BayesB did not outperform genomic best linear unbiased prediction (GBLUP). In the third study, we analyzed two backcross (BC) families derived from a resistant and a susceptible recurrent parent. Both BC populations differed substantially in their means for all traits, suggesting that the two recurrent parents have different QTL alleles for GER resistance. Relatively high correlations were observed between DON and ZON concentrations measured by immunoassays and GER visual severity scoring and NIRS (near-infrared spectroscopy) within each BC population. Thus, the mycotoxin content in grain can reliably be reduced by directional selection for GER severity and NIRS measurements that are less expensive and less laborious. In conclusion, GER resistance in European maize germplasm can be effectively improved through breeding with resistant donor lines. GER visual severity scoring and NIRS measurements were found to be reliable predictors for DON and ZON concentrations in grain. We observed that QTL for GER and DON are mostly specific to a few families or a limited number of materials, whereas QTL for DS are more commonly shared between families. The multi-parental QTL mapping approach is complementary to the classical bi-parental QTL mapping in that the latter has generally higher power to identify rare but large-effect QTL for traits such as GER and DON, whereas the former is superior in detecting common but small-effect QTL for traits such as DS. Composing the TS with materials more closely related to the prediction set and increasing the TS size generally resulted in higher prediction accuracy for MAS and GS, irrespective of the trait and statistical model.
Inheritance of Barley yellow dwarf virus resistance in maize
(2015) Horn, Frederike; Stich, Benjamin
Barley yellow dwarf (BYD) is one of the economically most important virus diseases in cereals. Due to increasing winter temperatures it is expected that BYD will become an increasing problem in maize cultivation. In earlier studies, it was reported that BYD has a negative impact on plant performance of maize. BYD virus (BYDV) is transmitted by aphids and the best control of the virus is the development of resistant maize cultivars. Therefore, the first objectives of my thesis research were to (i) determine phenotypic and genotypic variation in five segregating populations and in a broad germplasm set of maize with respect to BYDV tolerance and resistance as well as to (ii) quantify the influence of BYDV infection on the plant traits plant height, ear height, and flowering time. I observed a negative impact of BYDV infection on maize plant traits which shows that the development of resistant maize cultivars is of high importance for maize cultivation. Furthermore, in the connected biparental populations as well as in the association mapping population, I observed a high genotypic variance with regard to BYDV resistance which is the requirement for successful breeding and the identification of genome regions which contribute to BYDV resistance. The evaluation of BYDV resistance by the inoculation with BYDV and by double antibody sandwich enzyme-linked immunosorbent assay (DASELISA) is dificult to be included in the breeding process. Therefore, molecular markers are of high importance for the improvement of BYDV resistance by breeding. Therefore, the objective of this study was the (iii) identification of genome regions which are involved in the BYDV resistance by a genome wide association study (GWAS). For the BYDV resistance traits, significantly (α=0.01) associated SNPs were identified in the GWAS on chromosome 10 and 4. The SNPs identified for virus extinction on chromosome 10 explained in a simultaneous fit 25% of the phenotypic variance and were located in gene regions which were in other plants described to be involved in resistance mechanisms. This suggests that BYDV resistance is inherited oligogenically and that genes involved in general resistance mechanisms are also involved in BYDV resistance in maize. GWAS has the advantage that a large number of alleles per locus can be surveyed simultaneously, and because historical recombinations can be used, the mapping resolution is higher compared to classical linkage mapping. Nevertheless, genes contributing to phenotypic variation which show a low allele frequency can remain undetected. Due to a balanced allele frequency in segregating populations, linkage mapping has the advantage of higher QTL detection power compared to GWAS. Therefore, the objective of this study was to (iv) validate the genome regions with a linkage analysis in connected biparental crosses. The genome region on chromosome 10 which was identified in the GWAS to be linked to BYDV resistance could be validated in the linkage mapping study with connected populations as well as in the single populations. Furthermore, the QTL on chromosome 10 colocalized with the QTL identified in controlled greenhouse conditions. In earlier studies, QTL for other virus resistances were identified on chromosome 10. This suggests that these genes are involved in multiple virus resistances. The identified genome regions explain 45% of the phenotypic variance and are, therefore, promising for the use in MAS. The broad genotypic variation with regard to BYDV resistance, observed in my thesis research, provided a good basis for the successful identification of molecular markers which are associated with BYDV resistance in maize. The markers identified in my study by GWAS were validated by a linkage mapping approach and are promising for the use in marker assisted selection on BYDV resistance in maize breeding.
Joint QTL analysis of three connected F2-crosses in pigs
(2012) Rückert, Christine; Bennewitz, Jörn
Mapping Quantitative Trait Loci (QTL) has received considerable attention in livestock genetic research over the last two decades. Knowledge of the location, the mode of inheritance and the size of effects of QTL contribute to a deeper understanding of the genetic architecture of quantitative or complex traits. Furthermore, mapped QTL were envisaged for use in so-called marker assisted selection programs. Before the era of genomics started, microsatellites were usually used as genetic markers for QTL mapping. In pigs, F2-crosses were frequently established from divergently selected founder breeds. Usually, the sizes of these F2-experiments are in the range of 300 individuals, which is too small to obtain sufficient statistical power to map QTL precisely. One large F2-experiment was set up in the 90th of the last century at the University of Hohenheim. Three F2-crosses from three genetically different founder breeds (Meishan, Pietrain and European Wild Boar) with almost 1000 individuals were genotyped and phenotyped for around 50 quantitative traits. In further studies, each of the crosses were analysed separately and more complex modes of inheritance were ignored. However, several researchers showed that a combined analysis with several QTL experiments can boost statistical power. Additionally, the mode of inheritance is sometimes not restricted to additive and dominant gene action. The overall aim of this thesis was the joint analysis of these three F2-crosses with more appropriate statistical models and to draw more precise conclusions about the QTL segregating within these experimental designs.
Mapping of quantitative-trait loci (QTL) for adult-plant resistance to Septoria tritici in five wheat populations (Triticum aestivum L.)
(2010) Risser, Peter; Miedaner, Thomas
Septoria tritici blotch (STB), caused by Septoria tritici (teleomorph Mycosphaerella graminicola), is one of the most important diseases in wheat varieties worldwide, responsible for severe damage of the leaves causing yield losses between 30 and 40 %. Control of STB includes crop rotation, soil tillage, fungicide application, and cultivation of resistant varieties. Profit-making wheat growers are forced to apply narrow crop rotations under reduced tillage. Some fungicides including widely-used strobilurins are no longer effective due to mutations in the highly variable pathogen population of S. tritici. Therefore, resistance breeding using genetic mapping to identify quantitative-trait loci (QTL) associated with STB resistance provides a promising strategy for controlling the disease. The main goal of this study was to detect chromosomal regions for quantitative adult-plant resistance of winter wheat to STB. Besides this, we analyzed the genetic diversity of 24 European varieties after inoculation with four different isolates of S. tritici. Multienvironmental field trials inoculated with S. tritici were applied to test isolates and varieties and to phenotype mapping populations. In detail, the objectives were to (1) compare natural infection and inoculation, (2) evaluate genotypic variation of adult-plant resistance to STB in European varieties, (3) analyze genotype x environment (G x E) interaction, (4) evaluate and analyze phenotypic data including STB severity, heading date (HED), and plant height (PLH) of five mapping populations, (5) construct genetic linkage maps of these populations using AFLP, DArT, and SSR markers, (6) determine number, positions, and genetic effects of QTL for evaluated traits, and (7) reveal QTL regions for multiple-disease resistance within mapping populations using QTL meta-analysis. In all trials, inoculation with one to four preselected isolates was performed and STB severity was visually scored plotwise as percentage coverage of flag leaves with lesions bearing pycnidia. 24 winter wheat varieties were chosen with maximal differentiation in resistance to STB and evaluated across three years including nine environments. Five mapping populations, Florett/Biscay, Tuareg/Biscay, History/Rubens, Arina/Forno, and Solitär/Bussard, each comprising a cross of a resistant and a susceptible variety, with population sizes ranging from 81 to 316, were phenotyped across four to six environments. In parallel, 221 to 491 polymorphic genetic markers were assigned to linkage groups covering 1,314 to 3,305 cM of the genome. Based on these linkage maps, the number, positions, and genetic effects of QTL could be determined by composite interval mapping. Furthermore, raw data of different experiments evaluated for resistance to two other pathogens, Fusarium head blight and Stagnospora glume blotch, were used to reveal multiple-disease resistance QTL within Arina/Forno and History/Rubens populations by the software package PLABMQTL. Results of inoculated field trials coincided with not inoculated trials showing natural infection (r = 0.84 to 0.99, P < 0.01), thus inoculation method was accurate to evaluate STB severity in the field. Genotypic variation between 24 varieties ranged from 8 % (Solitär) to 63 % (Rubens) flag leaf area infected. In the analysis of variance, genotypic variance had highest impact followed by G x E interaction (P < 0.01). Therefore, environmental stability of varieties should be a major breeding goal. The varieties Solitär, History, and Florett were most stable, as revealed by a regression approach. In contrast, disease symptoms of Biscay ranged from 19 to 72 % within the three experimental years. Phenotypic data revealed significant (P < 0.01) genotypic differentiation for STB, HED, and PLH within all five mapping populations and between the parents. Entry-mean heritabilities (h²) ranged from 0.69 to 0.87 for STB, the only exception was Tuareg/Biscay (h² = 0.38). For HED (h² = 0.78 to 0.93) and PLH (h² = 0.92 to 0.98) heritabilities were high. All correlations between STB and HED (r = -0.18 to -0.33) as well as between STB and PLH (r = -0.13 to -0.45) were negative and moderate. The exception was History/Rubens which is segregating at the Rht-D1 locus showing considerably higher correlation between STB and PLH (r = -0.55, P < 0.01). The five mapping populations showed a wide and continuous distribution of mean STB severity averaged across three to six environments in field trials at adult-plant stage. In QTL analysis, one to nine, zero to nine, and four to eleven QTL were detected for STB, HED, and PLH, respectively, across five wheat populations using composite interval mapping. One to two major QTL for resistance to STB were detected consistently across environments in each population (QStb.lsa_fb-3B, QStb.lsa_fb-6D, QStb.lsa_tb-4B, QStb.lsa_tb-6B, QStb.lsa_hr-4D, QStb.lsa_hr-5B.1, QStb.lsa_af-3B, QStb.lsa_bs-7A) explaining more than 10 % of normalized adjusted phenotypic variance. Altogether, resistance QTL explained 14 to 55 % of adjusted phenotypic variance. Both parents contributed resistant alleles. Major QTL, however, were all from the resistant parent. QTL meta-analysis revealed each of four loci for multiple-disease resistance located on chromosomes 3B, 4B, 5B, and 6D in Arina/Forno, and on chromosomes 2B, 4D, 5B, and 7B in History/Rubens. The most effective meta QTL was on chromosome 4D in History/Rubens closely linked to Rht-D1. The resistance allele from History reduced disease severity by 9.8 % for STB and 6.3 % for FHB, thus explaining 47 % and 60 % of partial phenotypic variance. In general, European wheat varieties showed a wide range of genotypic variation for STB resistance useful for breeding. Although the influence of environment and G x E interaction was high, some resistant varieties which were stable across multiple environments were found (Solitär, History, Florett). Genomic regions associated with STB resistance were mapped across 13 out of 21 wheat chromosomes. Together with the continuous distribution of five segregating populations for flag leaf infection, it can be concluded that the adult-plant resistance to S. tritici was inherited quantitatively depending on several loci explaining part of phenotypic variance. QTL meta-analysis across three severe pathogens, including Fusarium head blight, Stagnospora glume blotch, and STB, within two populations revealed eight loci for multiple-disease resistance with closely linked markers applicable in resistance breeding. Combining detected major QTL as well as meta QTL in present breeding material by applying marker-assisted selection seems a promising approach to the breeding of varieties with improved resistance to Septoria tritici blotch, Fusarium head blight, and Stagnospora glume blotch.
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.
Markergestützte Vererbungsanalyse der Pollenfertilitätsrestauration bei Winterroggen (Secale cereale L.)
(2001) Wolf, Markus; Geiger, Hartwig H.
In rye (Secale cereale L.) as in a number of other crossfertilized crops, hybrid breeding allows a targeted use of heterosis for improving yield and yield stability. The creation of hybrids in outbred rye requires an efficient system of pollination control because of the monoclinous inflorescence. In commercial seed production, mostly the Pampa (P-) Cytoplasma is used, which induces cytoplasmatic male sterility (CMS). Restoration of pollen fertility is achieved by means of pollinator lines carrying nuclear dominant restorer genes. To obtain information about the number, localization, and efficacy of the restorer genes, molecular marker based inheritance analysis in rye was performed. The results were compared to already localized restorer sources in rye and other species. Ninety-two marker loci were mapped by RFLP technique, which covered a genome-length of 845 cM. QTL (quantitative trait loci) analysis revealed a major gene on the short arm of chromosome 1R. Two RFLP markers, psr596 and bcd1124, were linked with a distance of about 0.3 cM to this major gene. Moreover, two modifying minor genes on chromosome arms 3RL and 5RL were detected. One of these minor genes showed epistatic interaction with the above mentioned major gene.
Molecular and phenotypic analyses of pathogenicity, aggressiveness, mycotoxin production, and colonization in the wheat-Gibberella zeae pathosystem
(2004) Cumagun, Christian Joseph R.; Miedaner, Thomas
Fusarium head blight (FHB), caused by Gibberella zeae (Schwein.) Petch (anamorph: Fusarium graminearum Schwabe), is one of the principal diseases responsible for extensive damage in wheat fields and contamination of grain with the mycotoxins deoxynivalenol (DON) and nivalenol (NIV), rendering the harvest unsafe for human and animal consumption. Control of FHB is difficult because of the complex nature of host-pathogen-environment interaction and the nonavailability of highly effective fungicides. Agronomic practices and resistance breeding, therefore, offer the best strategies for disease management. Mapping by molecular markers provides an accurate approach for genetic analyses of simple and complex traits particularly pathogenicity, aggressiveness, and mycotoxin production. Pathogenicity, as defined here, is the ability to cause disease whereas aggressiveness is the quantity of disease induced by a pathogenic isolate on a susceptible host in which isolates do not interact differentially with host cultivars. The project aims to (1) map pathogenicity and aggressiveness of G. zeae based on a published genetic map (2) estimate genetic diversity of four parent isolates by PCR-based markers (3) examine the inheritance of pathogenicity, aggressiveness, mycotoxin type (DON/NIV), and DON production on a phenotypic basis, (4) analyse genetic covariation among aggressiveness, DON, and fungal colonization, (5) and compare aggressiveness of 42 isolates in greenhouse and field environments. Two crosses of G. zeae using nit (nitrate nonutilizing) marker technique were performed: (1) pathogenic DON-producing Z-3639 (Kansas, USA) x nonpathogenic NIV-producing R-5470 (Japan) belonging to lineage 7 and 6, respectively, and (2) DON-producing FG24 (Hungary) x FG3211 (Germany), both aggressive lineage 7 isolates. For the first cross, 99 progeny segregated in a consistent 61:38 for pathogenicity: nonpathogenicity in a two-year greenhouse experiment. Among the 61 pathogenic progeny, disease severity, measured as percentage infected spikelets, varied significantly (P = 0.01). Heritability for aggressiveness was high. Pathogenicity locus was mapped on linkage group IV near loci PIG1 (red pigment production), TOX1 (trichothecene toxin amount), and PER1 (perithecial production) explaining 60%, 43%, and 51% of the phenotypic variation, respectively. Two large aggressiveness QTLs were mapped on linkage group I linked to the locus TRI5 (trichodiene synthase in the trichothecene gene cluster) and an amplified fragment length polymorphism (AFLP) marker (EAAMTG0655K), explaining 51% and 29% of the observed phenotypic variation, respectively. These unlinked loci suggest that genetic basis between pathogenicity and aggressiveness were different. TRI5 is located in the same gene cluster as a previously identified gene known as TRI13, which determines whether DON or NIV will be produced. DON-producing progeny were, on average, twice as aggressive as were those producing NIV. Loci were only detected in the two linkage groups mentioned from the nine linkage groups present in the map. For the second cross FG24 x FG3211 with 153 progeny, head blight rating and relative plot yield were used as aggressiveness traits. DON production was measured by a commercial kit enzyme immunoassay. These three traits were quantitatively inherited among 153 progeny across three environments. Repeatabilities within each environment were medium to high but heritabilities across environments were medium only due to high progeny-environment interaction. DON was a less environmentally stable trait than aggressiveness. Transgressive segregants were detected frequently. This implies that even a cross within a lineage could lead to an increase in aggressiveness. Mapping of this cross was not initiated because the parents were not polymorphic enough to construct a genetic map. Instead, the parents were analysed for polymorphism in comparison to the parents of the first cross using 31 AFLP primer combinations and 56 random amplified polymorphic DNA (RAPD) primers. Polymorphism between Z-3639 and R-5470 was about three to four times higher than between FG24 and FG3211. Cluster analysis revealed that R-5470 was genetically separated from the other three parents, thus confirming the lineage assignments. Among preselected 50 progeny from the same field experiments that showed normal distribution for aggressiveness - head blight rating, fungal colonization, and DON production were correlated (r = 0.7, P = 0.01). Fungal colonization measured as Fusarium exoantigen (ExAg) content using enzyme-linked immunosorbent assay (ELISA) varied also quantitatively, but heritability was lower due to high progeny-environment interaction and error. Strong correlations among all traits indicate control by similar genes or gene complexes. No significant variation was observed for DON/ExAg ratio. Aggressiveness traits and DON production were more environmentally stable compared to Fusarium ExAg content. Our findings imply that aggressiveness may have other components apart from mycotoxin production. Genotypic variation for aggressiveness among the 42 progeny in one greenhouse and three field environments was significant and their correlation was moderate (r = 0.7, P = 0.01). High heritability in both environments again indicates that aggressiveness was a relatively stable trait, although methods of inoculation differed, i.e., injection for greenhouse and spraying for field experiments. Greenhouse aggressiveness could predict aggressiveness in the field, and thereby should reduce costs for resistance and phytopathological studies. In conclusion, we consider G. zeae as medium-risk pathogen with the potential to evolve to a higher level of aggressiveness due to sexual recombination. Erosion of quantitative resistance in FHB cannot be ignored, especially if host resistances with oligogenic inheritance, e.g. Sumai 3 from China, are used on a large acreage. Consequently, the rather simple inheritance of pathogenicity and aggressiveness in G. zeae could lead to a gradual increase of aggressiveness. These results should enhance efforts of plant breeders to use several, genetic distinct sources of resistance in order to avoid possible FHB outbreaks in the future.
Multiomics based association mapping in wheat reveals genetic architecture of quality and allergenic related proteins
(2023) El Hassouni, Khaoula; Afzal, Muhammad; Steige, Kim A.; Sielaff, Malte; Curella, Valentina; Neerukonda, Manjusha; Tenzer, Stefan; Schuppan, Detlef; Longin, Carl Friedrich Horst; Thorwarth, Patrick
Wheat is an important staple crop since its proteins contribute to human and animal nutrition and are important for its end-use quality. However, wheat proteins can also cause adverse human reactions for a large number of people. We performed a genome wide association study (GWAS) on 114 proteins quantified by LC-MS-based proteomics and expressed in an environmentally stable manner in 148 wheat cultivars with a heritability > 0.6. For 54 proteins, we detected quantitative trait loci (QTL) that exceeded the Bonferroni-corrected significance threshold and explained 17.3–84.5% of the genotypic variance. Proteins in the same family often clustered at a very close chromosomal position or the potential homeolog. Major QTLs were found for four well-known glutenin and gliadin subunits, and the QTL segregation pattern in the protein encoding the high molecular weight glutenin subunit Dx5 could be confirmed by SDS gel-electrophoresis. For nine potential allergenic proteins, large QTLs could be identified, and their measured allele frequencies open the possibility to select for low protein abundance by markers as long as their relevance for human health has been conclusively demonstrated. A potential allergen was introduced in the beginning of 1980s that may be linked to the cluster of resistance genes introgressed on chromosome 2AS from Triticum ventricosum. The reported sequence information for the 54 major QTLs can be used to design efficient markers for future wheat breeding.
Phänotypische und molekulare Analyse von Kreuzungsnachkommen auf Resistenz gegen Ährenfusariosen bei Triticale (x Triticosecale Wittmack)
(2011) Großmann, Maren; Großmann, Maren
Fusarium head blight (FHB) is an epidemic disease of cereals but disease control still is insufficiently possible. Resistance breeding can be addressed as one approach to reduce the mycotoxin contents. Since 2006 the European Union had strict rules governing the values for Desoxinivalenol (DON) in food products. The extraordinary loads through mycotoxins in triticale, which are caused by FHB, lead to large reductions in grain yield and quality. Beyond the fact that it can cause health issues in animals, especially in pig feed, it may induce financial burdens for farming industries. Resistance against FHB is inherited quantitatively with mainly additive effects and therefore is only recorded by complex field experiments. Molecular markers accelerate resistance breeding and enable specific introgressions of favorable QTL. The main topic of this thesis is to show the application possibilities of molecular markers for the investigation of FHB resistance in triticale. In detail this study aims to analyze factors leading to a minimized mycotoxin accumulation in perennial field trials at several locations. Furthermore genetic maps of two different crossing populations were compiled and QTL mapping for ear appearance, plant height, DON content and FHB resistance was implemented. For all populations significant varieties had been displayed for all characteristics. The average values for FHB rate were between 8 and 43 % depending on year and location. The heritability?s showed high values for each population (h2 = 0.7 ? 0.82). The substantial genotype-environment-interaction pointed out the importance of field experiments. Furthermore no significant correlation was obtained between ear appearance and plant height. Due to the fact that the correlation factor between DON-content and FHB rate was very low (r = 0.32) to moderate (r = 0,65). Therefore no assumption about the DON-content subjected to the FHB rate could be made. The mapping of both populations LASKO x ALAMO and LASKO x TRIMESTER has been accomplished with SSR- and DArT-markers. The rate of polymorphism could be increased from 7 - 12 % respectively to 9 - 18 %. Genetic maps were constructed with length of 1.815 and 1.407 cM. They indicated 5.19 and 4.54 cM in their average marker distances. Throughout the QTL mapping several QTL were detected (6 QTL for ear appearance, 4 QTL for plant height, 1 QTL for DON-content and 4 QTL for FHB rate). These QTL explained 8 to 66 % of the phenotypical variance. In addition crossing populations of LASKO x TRIMESTER showed 2 QTL for ear appearance, 1 for plant height and 2 QTL for FHB resistance, which could explain 3 to 41 % of the phenotypical variance. This was also observed in several other QTL-surveys for wheat. Prospectively using molecular markers and genetically engineered methods will increase the research and development of resistant varieties and lead to a reduced mycotoxin accumulation. So far neither genetic maps nor any QTL studies for FHB resistance or DON-content in triticale have been published. Due to the fact that resistance is transmitted by several genes numerous artificial inoculations have to be carried out during breeding. Additive gene effects are useful to combine several resistance genes from different parents. Established methods in plant breeding such as recurrent selection, phenotypical and marker-based selection are successfully in use to reduce FHB- symptoms and to reduce the mycotoxin value but these methods have to be improved. The results of this thesis are promising for a superior resistance breeding in the future.
Phenotypic and molecular analyses of grain and biomass productivity under irrigated and rainfed conditions in hybrid rye
(2014) Gottwald, Marlen; Miedaner, Thomas
Rye (Secale cereale L.) is a small grain cereal used for bread making, livestock feeding and as renewable energy source. These types of usages are leading to different breeding goals. Rye growing regions are affected by climate change and consequently by drought. Germany is touched by rainless periods in spring and early summer in the last years. Again, in spring 2012 farmers in Brandenburg and Lower Saxony were affected by drought periods. Yield losses in those regions, especially in combination with sandy soils are expected. Therefore much attention is paid for breeding of drought resistant germplasm. Briefly, our objectives of this study were to (1) estimate the biomass and biogas potential of different plant materials, their quantitative genetic parameters and biogas-related traits, (2) analyze two recombinant inbred lines and differences in their yield potential between irrigated and rainfed regime, as well as the relative efficiency for indirect selection for drought resistance in irrigated regime, and (3) investigate the phenotypic performance for ten agronomic and quality traits across multiple environments and estimated the number and effects underlying QTL. For the biomass-/ biogas analyses a wide range of plant material was analysed. Germplasm resources, full-sib families selected for grain and forage use were tested for their per se and testcross performance and experimental hybrids selected for grain use and population cultivars selected for grain and forage use were analyzed. Dry matter yields varying across environments from 106 to 177 dt/ha for per se and testcross performance, respectively. For testcross performance, germplasm resources showed similar values to forage rye. The later the maturity stage, the more dry matter yield on the whole plant level was achieved. Estimates of genotypic variances for biomass yield were significant for all rye materials, whereas the variances per se and for testcrosses were for germplasm resources exorbitant higher than for forage and grain rye. Typical cumulative methane production curves were obtained for the whole plant material from the Hohenheim biogas yield test. Methane yield showed large differences between second and third harvest date for individual plant fractions. Differences between genotypes were not substantial for methane yield although significant in some instances. At EC77/83 hybrids and forage rye reached similar methane yield of about 5000 m3/ha. A high correlation between dry matter yield and methane yield was observed (r=0.95). Concerning high cost and time consuming analysis of biogas tests, for breeders the main breeding goal should be maximum dry matter yield. Direct selection on dry matter yield should indirect improve methane yield. Two biparental populations were used for the analysis of drought tolerance. The analysis was performed in duplicate. Both populations were grown under irrigated and rainfed regimes. Striking less rainfall compared to long-term precipitation occurred between April and July, during critical phases of plant development. Grain yield reduction between irrigated and non-irrigated regime ranged from 2% to 29.6% for population A and 2% to 40% for population B, whereas differences between both regimes were significant (P<0.05) for five and four environments, respectively. Genotypic variances of grain yield were significant in all instances, whereas genotype by irrigation interaction variance between both regimes being significant only in three and four environments for population A and B, respectively. Analysis across those environments revealed significant difference for genotype by irrigation interaction variance and the three-way interaction variance in both populations. Heritability estimates were higher for the irrigated than for the rainfed regime. High interaction variance with environment and no clustering of the two regimes in a multi-dimensional analysis were found. This illustrates the different soil and whether conditions between locations and additionally every location suffered from a different drought stress. The correlation between both regimes was significant but moderate, but genotypic coefficients considerably higher (Pop-A: 0.86, Pop-B: 0.84), which could be substantiated that testcrosses differed not substantially in drought-resistance. Indirect selection for drought in the irrigated regime was predicted to be equally or more efficient than direct selection in the non-irrigated regime. Phenotypic and genotypic analysis was done across ten environments for both biparental populations for the general improvement of agronomic and quality traits in rye. Population A were genotyped with a Rye5K SNP array and for population B DArT genotyping was done with a 3K rye array. Additionally both populations were genotyped with about 150 SSRs. The genetic linkage maps comprised 1,819 and 1,265 markers for population A and B, respectively and were used for the QTL analysis for ten agronomic and quality traits. Phenotyping revealed large genetic variation for ten agronomic and quality traits. Intensive phenotyping at up to ten environments led to moderate to high heritabilities. Across environments explained genotypic variance of the individual QTL ranged from 5 to 55%. For 1000-kernel weight, test weight, falling number, and starch content, several QTL with high effects and a frequency of recovery of about 90% were identified in both population. Rye suffered from drought stress in the last decade. Focusing on general improvement of rye regarding yield and quality, as well as improving rye regarding drought-resistance is important. Future research should be done in fine mapping and validation of the detected QTLs, for exploiting their potential in marker assisted breeding.
Phenotypic, genetic, and genomic assessment of triticale lines and hybrids
(2017) Losert, Dominik; Würschum, Tobias
Triticale (×Triticosecale Wittmack) is a small grain cereal used for livestock feeding and as renewable energy source. These diverse types of usage lead to different breeding strategies, ideally resulting in continued increase of both, grain and biomass yield. Briefly, the objectives of this thesis were to explore aspects with relevance for line and hybrid breeding in triticale by phenotypic, genetic and genomic assessment of important traits. More specifically, the objectives of this study were to (i) evaluate agronomic traits, assess trait correlations, and investigate the amount of heterosis in triticale hybrids, (ii) examine the potential of line and hybrid cultivars for production of biomass, (iii) assess the phenotypic and genotypic variability in triticale germplasm, (iv) investigate long-term phenotypic trends based on cultivars registered in the past three decades, and (v) identify QTL for agronomical relevant traits. In conclusion, hybrids of triticale possess an increased biomass yield potential compared with their mid-parent values as well as compared with commercial reference cultivars. The findings on triticale germplasm and its breeding history provide important information for breeding programs. Furthermore, based on the obtained results, genomic approaches like marker-assisted or genomic selection appear promising to assist triticale breeding in the future.
QTL mapping and genomic prediction of complex traits based on high-density genotyping in multiple crosses of maize (Zea mays L.)
(2013) Stange, Michael; Melchinger, Albrecht E.
Most important agronomic traits like disease resistance or grain yield (GY) in maize show a quantitative trait variation and, therefore, are controlled by dozens to thousands of quantitative trait loci (QTL). Mapping of these QTL is well established in plant genetics to elucidate the genetic architecture of quantitative traits and to detect QTL for knowledge-based breeding. Nowadays, high-density genotyping is routinely applied in maize breeding and offers a huge number of SNP markers used in association mapping and genomic selection (GS). This enables also the construction of high-density linkage maps with marker densities of 1 cM or even higher. Nevertheless, QTL mapping studies were until recently mostly based on low-density maps. This raises the question if high-density maps are an overkill for QTL mapping, or in contrast, if important QTL mapping parameters would profit from them. High-density maps could also be beneficial for dissection of the complex trait GY into its components 100-kernel weight (HKW) and kernel number (KN). Analysis of these less complex traits may help to unravel the genetic architecture and improve the predictive ability for complex traits. However, an open question is whether consideration of component traits and epistatic interactions in QTL mapping models are beneficial for predicting the performance of untested genotypes for the complex trait GY. In this thesis, high-density linkage maps were constructed for biparental maize populations of doubled haploid (DH) lines and applied in different QTL linkage mapping approaches. In detail, the objectives of this study were to (1) investigate the effect of high-density versus low-density linkage maps in QTL mapping of important QTL mapping parameters and to analyze the resolution of closely linked QTL with experimental data and computer simulations, (2) map QTL for HKW, KN, and GY with high-density maps and to analyze epistatic interactions, (3) compare the prediction accuracy for GY with different QTL mapping models, and (4) answer the question how the composition of the test set (TS) influences the accuracy in genomic prediction of progenies from individual crosses. This thesis was based on five interconnected biparental populations with a total of 699 DH lines evaluated in field experiments for GER resistance related traits as well as for HKW, KN, and GY. All DH lines were genotyped with the Illumina MaizeSNP50 Bead Chip and high-density linkage maps were constructed separately for each population. For evaluation of high-density versus low-density maps on QTL mapping parameters, three linkage maps with marker densities of 1, 2, and 5 cM were constructed, starting from the full linkage map with 7,169 markers mapped in the largest population (N=204). QTL mapping was performed with all three marker densities in the experimental population for GER resistance related traits and for yield related traits, as well as in a simulation study with different population sizes. In the simulation study, independent QTL with additive effects explaining 0.14 to 7.70% of the expected phenotypic variance, as well as linked QTL with map distances of 5 and 10 cM, were simulated. Results showed that high-density maps had only minor effects on the QTL detection power and the proportion of genotypic variance explained. In contrast, support interval length decreased with increasing marker density, indicating an increasing precision of QTL localization. The precision of QTL effect estimates was measured as deviation between the reference additive effects and the estimated QTL effects. It gained from an increase in marker density, especially for small and medium effect QTL. Increasing the marker density from 5 to 1 cM was advantageous for separately detecting linked QTL in coupling phase with both linkage distances. In conclusion, this study showed that QTL mapping parameters relevant for knowledge-based breeding profited from an increase in marker density. For QTL mapping of the complex trait GY and the components HKW and KN, three QTL mapping models were applied to the four largest populations, of which two models were based on the component traits HKW and KN. All models included tests for epistatic interactions. The results showed that heritability was slightly higher for the component traits compared to the complex trait. The average length of support intervals of detected QTL was short with 12 cM, indicating high precision of QTL localization. Co-located QTL with same parental origin of favorable alleles were detected within populations for different traits and between populations for same traits, reflecting common QTL across populations. However, to finally confirm these common QTL, multi-population QTL mapping should be conducted. Based on the detected QTL, predictions for GY showed that epistatic models did not outperform the respective additive models. Nevertheless, component trait based models can be advantageous for identification of favorable allele combinations for multiplicative traits. For all five populations, the comparison of genetic similarities reflected the crossing scheme with full-sib families, half-sib families and unrelated families. The evaluation of prediction accuracies for different scenarios depended on the composition of the TS. Highest prediction accuracies were observed for DH lines within full-sib families, medium values if full-sib DH lines were replaced by half-sib DH lines, and lowest values if the TS comprised of DH lines from unrelated crosses. In conclusion, I found high-density linkage maps to be advantageous for linkage mapping in biparental DH populations by improving important QTL mapping parameters. Higher costs for high-density genotyping are by far compensated by these advantages. Dissecting the complex trait GY into its component traits HKW and KN by component trait based QTL mapping models revealed a complex genetic network of GY. Future research should focus on high-density consensus maps applied in multi-population QTL mapping to take advantage of the improved QTL detection power and to confirm common QTL across populations.