Septoria Tritici Blotch Resistance Research Articles

Field response and genetic variability of elite spring bread wheat (Triticum aestivum L.) genotypes for septoria tritici blotch under natural infection in Northwest Ethiopia

AbstractFungal diseases cause significant yield loss to wheat production. Septoria tritici blotch (STB), caused by the ascomycete fungus Zymoseptoria trtici, is one of the major fungal diseases affecting wheat production worldwide. In Ethiopia, STB is a severe problem that causes significant yield loss in high and mid‐altitude wheat‐growing areas. The use of resistant varieties is one of the sustainable disease management strategies, particularly for resource‐poor farmers in developing countries. Two hundred and fifty bread wheat genotypes were evaluated to identify septoria tritici resistant genotypes and estimate the extent of genetic variability for resistance to STB and other economically important traits using alpha lattice design under natural infestation in two STB hotspot environments. Analysis of variance revealed highly significant differences among genotypes, environment, and genotype × environment interaction for all traits measured. The genetic coefficient of variance was lower than the phenotypic coefficient of variance for all traits studied, and both test environments showed the influence of the environment on trait expression. High and moderate heritability values were observed for the septoria disease severity parameters, indicating that the STB resistance trait was less influenced by the environment. The days to heading and plant height were inversely correlated with disease severity. This suggests that genotypes with tall plant height and long maturity period could be resistant to septoria tritici blotch through escape mechanisms. Four of the genotypes, namely, G‐215, G‐255, G‐257, and G‐258, were found to be resistant across all locations. These and other promising genotypes will be used in future breeding programmes to select or develop high‐yielding and STB‐resistant bread wheat genotypes that can be deployed in septoria tritici blotch‐prone areas. Highly susceptible genotypes will also be used as controls for STB resistance breeding programmes.

Genome-wide association study for resistances to yellow rust, powdery mildew, and Septoria tritici blotch in cultivated emmer

Emmer is a progenitor of bread wheat and evolved in the Levant together with the yellow rust (YR), powdery mildew (PM) fungi, and a precursor of Zymoseptoria tritici causing Septoria tritici blotch (STB). We performed a genome-wide association mapping for the three disease resistances with 143 cultivated emmer accessions in multi-environmental trials. Significant (P < 0.001) genotypic variation was found with high heritabilities for the resistances to the two biotrophs and a moderate heritability for STB resistance. For YR, PM, and STB severity nine, three, and seven marker-trait associations, respectively, were detected that were significant across all environments. Most of them were of low to moderate effect, but for PM resistance a potentially new major gene was found on chromosome 7AS. Genomic prediction abilities were high throughout for all three resistances (≥ 0.8) and decreased only slightly for YR and PM resistances when the prediction was done for the second year with the first year as training set (≥ 0.7). For STB resistance prediction ability was much lower in this scenario (0.4). Despite this, genomic selection should be advantageous given the large number of small QTLs responsible for quantitative disease resistances. A challenge for the future is to combine these multiple disease resistances with better lodging tolerance and higher grain yield.

Exploring GWAS and genomic prediction to improve Septoria tritici blotch resistance in wheat

Septoria tritici blotch (STB) is a destructive foliar diseases threatening wheat grain yield. Wheat breeding for STB disease resistance has been identified as the most sustainable and environment-friendly approach. In this work, a panel of 316 winter wheat breeding lines from a commercial breeding program were evaluated for STB resistance at the seedling stage under controlled conditions followed by genome-wide association study (GWAS) and genomic prediction (GP). The study revealed a significant genotypic variation for STB seedling resistance, while disease severity scores exhibited a normal frequency distribution. Moreover, we calculated a broad-sense heritability of 0.62 for the trait. Nine single- and multi-locus GWAS models identified 24 marker-trait associations grouped into 20 quantitative trait loci (QTLs) for STB seedling-stage resistance. The seven QTLs located on chromosomes 1B, 2A, 2B, 5B (two), 7A, and 7D are reported for the first time and could potentially be novel. The GP cross-validation analysis in the RR-BLUP model estimated the genomic-estimated breeding values (GEBVs) of STB resistance with a prediction accuracy of 0.49. Meanwhile, the GWAS assisted wRR-BLUP model improved the accuracy to 0.58. The identified QTLs can be used for marker-assisted backcrossing against STB in winter wheat. Moreover, the higher prediction accuracy recorded from the GWAS-assisted GP analysis implies its power to successfully select superior candidate lines based on their GEBVs for STB resistance.

Measuring resistance in durum wheat-Zymoseptoria tritici interaction using aggressiveness quantitative traits

Phenotyping for quantitative resistance to Septoria tritici blotch (STB), one of the most economically devastating diseases of durum wheat (Triticum durum Desf.) in North African region, is a limiting factor for breeding progress. Here we tested main phenotypic markers of aggressiveness affecting host damage and pathogen reproduction reported in earlier studies for measuring differences in quantitative host resistance. A collection of 11 Tunisian durum wheat genotypes varying in resistance to the pathogen was inoculated by a highly aggressive and virulent isolate of Zymoseptoria tritici in a replicated field and controlled experiments, conducted in 2017-2018 at the experimental station of the Regional Field Crops Research Center of Beja in Tunisia. The severity of infection caused by Z. tritici isolate, including percentage of leaf area covered by chlorotic lesions (PLACCL), percentage of leaf area covered by necrosis (PLACN), and percentage of leaf area covered by pycnidia (PLACP) as well as latent period (LP) and pycnidia density (PYCdens) were assessed visually and quantified. The results showed that there were significant differences in all assessed traits on the tested genotypes and significant positive correlations were obtained between PLACN, PLACP and PYCdens. Similarly, there was a significant negative correlation between LP and these last traits. The PLACP was found as the more reliable trait to discriminate genotypes to different classes of resistance, consistent with the results obtained for the remaining traits. These data suggest that using both necrotic leaf area (PLACN) bearing pycnidia and PYCdens can improve selection for quantitative STB resistance.

Genome-Wide Association Study Reveals Novel Genetic Loci for Quantitative Resistance to Septoria Tritici Blotch in Wheat (Triticum aestivum L.).

Septoria tritici blotch, caused by the fungus Zymoseptoria titici, poses serious and persistent challenges to wheat cultivation in Ethiopia and worldwide. Deploying resistant cultivars is a major component of controlling septoria tritici blotch (STB). Thus, the objective of this study was to elucidate the genomic architecture of STB resistance in an association panel of 178 bread wheat genotypes. The association panel was phenotyped for STB resistance, phenology, yield, and yield-related traits in three locations for 2 years. The panel was also genotyped for single nucleotide polymorphism (SNP) markers using the genotyping-by-sequencing (GBS) method, and a total of 7,776 polymorphic SNPs were used in the subsequent analyses. Marker-trait associations were also computed using a genome association and prediction integrated tool (GAPIT). The study then found that the broad-sense heritability for STB resistance ranged from 0.58 to 0.97 and 0.72 to 0.81 at the individual and across-environment levels, respectively, indicating the presence of STB resistance alleles in the association panel. Population structure and principal component analyses detected two sub-groups with greater degrees of admixture. A linkage disequilibrium (LD) analysis in 338,125 marker pairs also detected the existence of significant (p ≤ 0.01) linkage in 27.6% of the marker pairs. Specifically, in all chromosomes, the LD between SNPs declined within 2.26–105.62 Mbp, with an overall mean of 31.44 Mbp. Furthermore, the association analysis identified 53 loci that were significantly (false discovery rate, FDR, <0.05) associated with STB resistance, further pointing to 33 putative quantitative trait loci (QTLs). Most of these shared similar chromosomes with already published Septoria resistance genes, which were distributed across chromosomes 1B, 1D, 2A, 2B, 2D, 3A,3 B, 3D, 4A, 5A, 5B, 6A, 7A, 7B, and 7D. However, five of the putative QTLs identified on chromosomes 1A, 5D, and 6B appeared to be novel. Dissecting the detected loci on IWGSC RefSeq Annotation v2.1 revealed the existence of disease resistance-associated genes in the identified QTL regions that are involved in plant defense responses. These putative QTLs explained 2.7–13.2% of the total phenotypic variation. Seven of the QTLs (R2 = 2.7–10.8%) for STB resistance also co-localized with marker-trait associations (MTAs) for agronomic traits. Overall, this analysis reported on putative QTLs for adult plant resistance to STB and some important agronomic traits. The reported and novel QTLs have been identified previously, indicating the potential to improve STB resistance by pyramiding QTLs by marker-assisted selection.

Correction to: Genome-wide association study for septoria tritici blotch resistance reveals the occurrence and distribution of Stb6 in a historic Swiss landrace collection

Septoria tritici blotch (STB), caused by the fungus Zymoseptoria tritici, is a major constraint in global wheat production. The lack of genetic diversity in modern elite wheat cultivars largely hinders the improvement of STB resistance. Wheat landraces are reservoirs of untapped genetic diversity, which can be exploited to find novel STB resistance genes or alleles. Here, we characterized 188 Swiss wheat landraces for resistance to STB using four Swiss Z. tritici isolates. We used a genome-wide association study (GWAS) to identify genetic variants associated with the amount of lesion and pycnidia production by the fungus. The majority of the landraces were highly resistant for both traits to the isolate 1E4, indicating a gene-for-gene relationship, while higher phenotypic variability was observed against other isolates. GWAS detected a significant SNP on chromosome 3A that was associated with both traits in the isolate 1E4. The resistance response against 1E4 was likely controlled by the Stb6 gene. Sanger sequencing revealed that the majority of these ~ 100-year-old landraces carry the Stb6 resistance allele. This indicates the importance of this gene in Switzerland during the early 1900s for disease control in the field. Our study demonstrates the importance of characterizing historic landrace collections for STB resistance to provide valuable information on resistance variability and contributing alleles. This will help breeders in the future to make decisions on integrating such germplasms in STB resistance breeding.

Detection of virulence to septoria tritici blotch (STB) resistance conferred by the winter wheat cultivar Cougar in the Irish Zymoseptoria tritici population and potential implications for STB control

AbstractSeptoria tritici blotch (STB) caused by the fungal pathogen Zymoseptoria tritici continues to be the most economically destructive disease of winter wheat throughout Ireland. Due to the widespread development of fungicide resistance in the Irish Z. tritici population, integrated strategies to control STB are increasingly necessary. A key component of such strategies will be the deployment of winter wheat cultivars with improved levels of STB resistance. Unfortunately, due to the nature of Z. tritici, such resistances are at risk of being overcome by the pathogen. In late summer 2020, foci of STB were observed across a range of winter wheat cultivars under evaluation for recommendation in Ireland. Common amongst these was the cultivar Cougar in each of their pedigree. To determine if the foci observed in 2020 resulted from strains virulent to Cougar, isolate collections were established and virulence screens conducted on Cougar and a range of the cultivars currently under evaluation. These confirmed the presence of Cougar‐virulent strains in the Irish Z. tritici population, and that this virulence affects not just Cougar, but also cultivars derived from it. Although the foci observed in 2020 were in both fungicide‐untreated and ‐treated plots, there was no evidence that these strains are more sensitive or resistant to fungicides compared to the wider Irish Z. tritici population, with moderate resistance to the SDHIs and azoles dominating. Combined, the present study confirms the need to ensure a diversity of control measures for STB, including ensuring a range of STB resistances are used.

Mapping for Adult-Plant Resistance Against Septoria Tritici Blotch in a Common Wheat Line Murga.

Septoria tritici blotch (STB) is a major foliar disease globally that is notorious for quickly developing fungicide resistance, making host resistance an indispensable component in mitigating STB. The International Maize and Wheat Improvement Center (CIMMYT) wheat line Murga is well known for its high, durable, and broad-spectrum resistance against STB infection. This study aimed to investigate the resistance mechanism of Murga to facilitate its utilization in breeding. A recombinant inbred line population was derived from a cross between Murga and STB-susceptible line Huirivis#1, comprising 297 progenies. The population was evaluated for adult-plant STB resistance in Toluca, Mexico (from 2017 to 2019), and in La Estanzuela, Uruguay (from 2016 to 2018). Genotyping was performed with the DArTseq platform. Quantitative trait locus (QTL) mapping indicated a major and stable QTL on chromosome 3DL, explaining a phenotypic variation for STB of 41.2 to 62.5% in Mexico and 27.5 to 40.3% in Uruguay. This QTL was regarded as Stb16 based on the comparison of its physical position, the possible origin from synthetic wheat, and its broad-spectrum resistance. Additional QTLs with minor effects were identified on chromosomes 2B, 2D, 3A, 3B, and 5B. The QTL on 5BS was significant in four of the six environments and must be new. Murga was the resistant donor for all QTLs except for those on 2B and 3A. Being an elite breeding line, Stb16 carrier Murga could be used as a promising STB resistance donor. Rational employment of Stb16 could contribute to STB management yet avoid the rapid emergence of Stb16-virulent isolates.

QTL Mapping of Seedling and Adult Plant Resistance to Septoria Tritici Blotch in Winter Wheat cv. Mandub (Triticum aestivum L.)

Septoria tritici blotch (STB) is one of the most devastating foliar diseases of wheat worldwide. Host resistance is the most economical and safest method of controlling the disease, and information on resistance loci is crucial for effective breeding for resistance programs. In this study we used a mapping population consisting of 126 doubled-haploid lines developed from a cross between the resistant cultivar Mandub and the susceptible cultivar Begra. Three monopycnidiospore isolates of Z. tritici with diverse pathogenicity were used to test the mapping population and parents’ STB resistance at the seedling stage (under a controlled environment) and adult plant stage (polytunnel). For both types of environments, the percentage leaf area covered by necrosis (NEC) and pycnidia (PYC) was determined. A linkage map comprising 5899 DArTSNP and silicoDArT markers was used for the quantitative trait loci (QTL) analysis. The analysis showed five resistance loci on chromosomes 1B, 2B and 5B, four of which were derived from cv. Mandub. The location of QTL detected in our study on chromosomes 1B and 5B may suggest a possible identity or close linkage with Stb2/Stb11/StbWW and Stb1 loci, respectively. QStb.ihar-2B.4 and QStb.ihar-2B.5 detected on chromosome 2B do not co-localize with any known Stb genes. QStb.ihar-2B.4 seems to be a new resistance locus with a moderate effect (explaining 29.3% of NEC and 31.4% of PYC), conferring resistance at the seedling stage. The phenotypic variance explained by QTL detected in cv. Mandub ranged from 11.9% to 70.0%, thus proving that it is a good STB resistance source and can potentially be utilized in breeding programs.

Identification of Resistance Sources and Genome-Wide Association Mapping of Septoria Tritici Blotch Resistance in Spring Bread Wheat Germplasm of ICARDA.

Septoria tritici blotch (STB) of wheat, caused by the ascomycete Zymoseptoria tritici (formerly Mycosphaerella graminicola), is one of the most important foliar diseases of wheat. In Morocco, STB is a devastating disease in temperate wheat-growing regions, and the yield losses can exceed up to 50% under favorable conditions. The aims of this study were to identify sources of resistance to STB in Septoria Association Mapping Panel (SAMP), which is composed of 377 advanced breeding lines (ABLs) from spring bread wheat breeding program of ICARDA, and to identify loci associated with resistance to STB at seedling (SRT) as well as at the adult plant (APS) stages using genome-wide association mapping (GWAM). Seedling resistance was evaluated under controlled conditions with two virulent isolates of STB (SAT-2 and 71-R3) from Morocco, whereas adult plant resistance was assessed at two hot spot locations in Morocco (Sidi Allal Tazi, Marchouch) under artificial inoculation with a mixture of STB isolates. At seedling stage, 45 and 32 ABLs were found to be resistant to 71-R3 and SAT-2 isolates of STB, respectively. At adult plant stage, 50 ABLs were found to be resistant at hot spot locations in Morocco. Furthermore, 10 genotypes showed resistance in both locations during two cropping seasons. GWAM was conducted with 9,988 SNP markers using phenotypic data for seedling and the adult plant stage. MLM model was employed in TASSEL 5 (v 5.2.53) using principal component analysis and Kinship Matrix as covariates. The GWAM analysis indicated 14 quantitative trait loci (QTL) at the seedling stage (8 for isolate SAT-2 and 6 for isolate 71-R3), while 23 QTL were detected at the adult plant stage resistance (4 at MCH-17, 16 at SAT-17, and 3 at SAT-18). SRT QTL explained together 33.3% of the phenotypic variance for seedling resistance to STB isolate SAT-2 and 28.3% for 71-R3, respectively. QTL for adult plant stage resistance explained together 13.1, 68.6, and 11.9% of the phenotypic variance for MCH-17, SAT-17, and SAT-18, respectively. Identification of STB-resistant spring bread wheat germplasm in combination with QTL detected both at SRT and APS stage will serve as an important resource in STB resistance breeding efforts.

TaWRKY10 transcription factor is a novel jasmonic acid signalling regulator involved in immunity against Septoria tritici blotch disease in wheat

Septoria tritici blotch (STB), caused by the fungus Zymoseptoria tritici, is currently the main threat to worldwide wheat production in temperate climates. Understanding the genetic mechanisms that underpin the Z. tritici–wheat interaction will be crucial for generating new control strategies against STB. Plant hormones are essential regulators of growth and immunity. In particular, jasmonic acid (JA) plays a central role in defence against necrotrophic plant pathogens, but its role in mediating immunity against Z. tritici is largely unknown. Here we identify the transcription factor TaWRKY10 that is taxonomically restricted to the grass subfamily Pooideae as a novel regulator of JA responses in wheat. We demonstrate by using virus‐induced gene silencing that silencing TaWRKY10 leads to increased resistance against Z. tritici with an earlier onset of necrotic symptoms, but with reduced pathogen sporulation. Wheat plants silenced for TaWRKY10 show an up‐regulated JA response. Transcriptional profiling of TaWRKY10 knock‐down plants indicates that it is a negative regulator of the JA receptor TaCOI1 gene expression. Our data indicate that TaWRKY10 down‐regulates JA responses, contributing to the susceptibility of wheat to Z. tritici. We postulate that manipulating TaWRKY10 may provide a strategy to boost STB resistance in wheat.

Genome-wide association study for septoria tritici blotch resistance reveals the occurrence and distribution of Stb6 in a historic Swiss landrace collection

Septoria tritici blotch (STB), caused by the fungus Zymoseptoria tritici, is a major constraint in global wheat production. The lack of genetic diversity in modern elite wheat cultivars largely hinders the improvement of STB resistance. Wheat landraces are reservoirs of untapped genetic diversity, which can be exploited to find novel STB resistance genes or alleles. Here, we characterized 188 Swiss wheat landraces for resistance to STB using four Swiss Z. tritici isolates. We used a genome-wide association study (GWAS) to identify genetic variants associated with the amount of lesion and pycnidia production by the fungus. The majority of the landraces were highly resistant for both traits to the isolate 1E4, indicating a gene-for-gene relationship, while higher phenotypic variability was observed against other isolates. GWAS detected a significant SNP on chromosome 3A that was associated with both traits in the isolate 1E4. The resistance response against 1E4 was likely controlled by the Stb6 gene. Sanger sequencing revealed that the majority of these ~ 100-year-old landraces carry the Stb6 resistance allele. This indicates the importance of this gene in Switzerland during the early 1900s for disease control in the field. Our study demonstrates the importance of characterizing historic landrace collections for STB resistance to provide valuable information on resistance variability and contributing alleles. This will help breeders in the future to make decisions on integrating such germplasms in STB resistance breeding.

Genetic Analysis Using a Multi-Parent Wheat Population Identifies Novel Sources of Septoria Tritici Blotch Resistance.

Zymoseptoria tritici is the causative fungal pathogen of septoria tritici blotch (STB) disease of wheat (Triticum aestivum L.) that continuously threatens wheat crops in Ireland and throughout Europe. Under favorable conditions, STB can cause up to 50% yield losses if left untreated. STB is commonly controlled with fungicides; however, a combination of Z. tritici populations developing fungicide resistance and increased restrictions on fungicide use in the EU has led to farmers relying on fewer active substances. Consequently, this serves to drive the emergence of Z. tritici resistance against the remaining chemistries. In response, the use of resistant wheat varieties provides a more sustainable disease management strategy. However, the number of varieties offering an adequate level of resistance against STB is limited. Therefore, new sources of resistance or improved stacking of existing resistance loci are needed to develop varieties with superior agronomic performance. Here, we identified quantitative trait loci (QTL) for STB resistance in the eight-founder “NIAB Elite MAGIC” winter wheat population. The population was screened for STB response in the field under natural infection for three seasons from 2016 to 2018. Twenty-five QTL associated with STB resistance were identified in total. QTL either co-located with previously reported QTL or represent new loci underpinning STB resistance. The genomic regions identified and the linked genetic markers serve as useful resources for STB resistance breeding, supporting rapid selection of favorable alleles for the breeding of new wheat cultivars with improved STB resistance.

Taxonomically Restricted Wheat Genes Interact With Small Secreted Fungal Proteins and Enhance Resistance to Septoria Tritici Blotch Disease.

Understanding the nuances of host/pathogen interactions are paramount if we wish to effectively control cereal diseases. In the case of the wheat/Zymoseptoria tritici interaction that leads to Septoria tritici blotch (STB) disease, a 10,000-year-old conflict has led to considerable armaments being developed on both sides which are not reflected in conventional model systems. Taxonomically restricted genes (TRGs) have evolved in wheat to better allow it to cope with stress caused by fungal pathogens, and Z. tritici has evolved specialized effectors which allow it to manipulate its’ host. A microarray focused on the latent phase response of a resistant wheat cultivar (cv. Stigg) and susceptible wheat cultivar (cv. Gallant) to Z. tritici infection was mined for TRGs within the Poaceae. From this analysis, we identified two TRGs that were significantly upregulated in response to Z. tritici infection, Septoria-responsive TRG6 and 7 (TaSRTRG6 and TaSRTRG7). Virus induced silencing of these genes resulted in an increased susceptibility to STB disease in cvs. Gallant and Stigg, and significantly so in the latter (2.5-fold increase in STB disease). In silico and localization studies categorized TaSRTRG6 as a secreted protein and TaSRTRG7 as an intracellular protein. Yeast two-hybrid analysis and biofluorescent complementation studies demonstrated that both TaSRTRG6 and TaSRTRG7 can interact with small proteins secreted by Z. tritici (potential effector candidates). Thus we conclude that TRGs are an important part of the wheat-Z. tritici co-evolution story and potential candidates for modulating STB resistance.

Identification of valuable sources of resistance to Zymoseptoria tritici in the Tunisian durum wheat landraces

Tunisia is one of the main producers of durum wheat, the most consumed cereal in Tunisia and represents a trademark of several local dishes such as couscous and bulgur. Nonetheless and despite its leading stand in the local consumption and commercial share, a scarcity in Tunisian durum wheat production has long been a major problem obstructing the fulfilment of the increasing local demand. Septoria tritici blotch (STB) caused by Zymoseptoria tritici (Z. tritici) is one of the most common and detrimental foliar disease on durum wheat in Tunisia causing considerable yield losses. To-date and despite the damaging effects of STB on durum wheat, limited sources of resistance to Z. tritici have been identified. In this present study, we assessed necrosis and pycnidia development of a collection of 304 Tunisian durum wheat landraces representative of 11 landrace populations and artificially inoculated with Z. tritici at the seedling stage under controlled conditions and at the adult plant stage under field conditions. Based on necrosis and pycnidia scores, a hierarchical classification analysis clustered the durum wheat landraces into three phenotypic classes of response to Z. tritici, namely resistant, intermediate and susceptible genotypes. While resistant plants represented the most frequent class at the seedling stage, susceptible phenotypes were more frequent at the adult plant stage. Nevertheless, a heat map correlation study showed that resistant landraces with low-percentages of necrosis and pycnidia at seedling and adult plant stages represented the dominant group within the tested collection. Moreover, the assessed frequencies per landrace populations of the resistant, the intermediate and the susceptible accessions to Z. tritici infection at the adult plant stage for pycnidia development has shown that the landrace populations Badri, Jneh Khotifa (JK), Mekki and Taganrog were the highest resistant accessions, in contrast to the landrace populations Biskri, Mahmoudi, Azizi and Biada where susceptible genotypes were predominant. Mahmoudi and Azizi represented the most variable landrace populations encompassing the three phenotypic classes. Our results highlight that Tunisian durum wheat landraces may harbour variable and stable sources of resistance to Z. tritici at seedling and adult plant stages, which would represent a good basis for further investigation and deployment in breeding programs to enhance STB resistance.

First report of virulence to the septoria tritici blotch resistance gene Stb16q in the Irish Zymoseptoria tritici population

In June 2019 high levels of septoria tritici blotch (STB), caused by the fungal pathogen Zymoseptoria tritici were observed in Recommended List evaluation plots of winter wheat cv. Cellule (Fig. 1) in the Republic of Ireland. Levels of STB were unexpected as Cellule has the STB resistance gene Stb16q and previous evaluations confirmed its high level of resistance (Table 1). Septoria tritici blotch is the most destructive disease of winter wheat in Western Europe and control is heavily reliant on fungicides (O'Driscoll et al., 3). Due to increased regulations on the use of fungicides in Europe varietal resistance is regarded as key to managing STB, with 21 resistance genes identified and mapped (Brown et al., 1). Amongst these, Stb16q is unique, providing high levels of resistance equally at both the seedling and adult plant stages (Ghaffary et al., 2). As such it is important to determine if the levels of disease observed in 2019 were due to the emergence of virulence to Stb16q. A collection of Z. tritici isolates were established from infected leaf samples of the Stb16q-resistant cv. Cellule (n=10) and the moderately susceptible cv. Costello (n=10). All isolates were from fungicide-untreated commercial crops or trial plots. To determine if those isolates retrieved from Cellule exhibited virulence on Cellule when compared to those isolated from Costello, glasshouse seedling assays were performed. Two-week old seedlings of both Cellule and the susceptible cv. KWS Lumos were inoculated with spore suspensions (106 spores/ml) of each isolate until runoff; placed in a clear polyethylene bag for 48 hr and subsequently moved into a glasshouse. Levels of disease were assessed 21 days post inoculation. The experiment was conducted twice. A beta generalised mixed-effects model (block and source cultivar as fixed and test cultivar as a random component) was fitted to the data and the estimated marginal means were separated using Tukey's Honest Significant Difference (α=0.05). All isolates infected the susceptible variety, with no differences observed between isolate collections (Fig. 2). Significant differences (P<0.0001) were observed between the collections in their ability to cause infection on Cellule, with almost no disease detected following inoculation with Costello isolates (Figure 2). High levels of disease were observed on Cellule following inoculation with the Cellule isolates, confirming the presence of virulence to Stb16q in the Irish Z. tritici population. Whilst the prevalence of virulence in the wider Z. tritici population is unknown, its presence in the population must be taken into account when considering cultivating varieties reliant on Stb16q as the source of STB resistance. The authors thank Eleanor O'Gorman, Seamus Kearney and Lucie-Ann Lamard for assistance and Teagasc (0154) for funding.

GWAS-Assisted Genomic Prediction to Predict Resistance to Septoria Tritici Blotch in Nordic Winter Wheat at Seedling Stage.

Septoria tritici blotch (STB) disease caused by Zymoseptoria tritici is one of the most damaging diseases of wheat causing significant yield losses worldwide. Identification and employment of resistant germplasm is the most cost-effective method to control STB. In this study, we characterized seedling stage resistance to STB in 175 winter wheat landraces and old cultivars of Nordic origin. The study revealed significant (p < 0.05) phenotypic differences in STB severity in the germplasm. Genome-wide association analysis (GWAS) using five different algorithms identified ten significant markers on five chromosomes. Six markers were localized within a region of 2 cM that contained seven candidate genes on chromosome 1B. Genomic prediction (GP) analysis resulted in a model with an accuracy of 0.47. To further improve the prediction efficiency, significant markers identified by GWAS were included as fixed effects in the GP model. Depending on the number of fixed effect markers, the prediction accuracy improved from 0.47 (without fixed effects) to 0.62 (all non-redundant GWAS markers as fixed effects), respectively. The resistant genotypes and single-nucleotide polymorphism (SNP) markers identified in the present study will serve as a valuable resource for future breeding for STB resistance in wheat. The results also highlight the benefits of integrating GWAS with GP to further improve the accuracy of GP.

QTL Mapping and Transcriptome Analysis to Identify Differentially Expressed Genes Induced by Septoria Tritici Blotch Disease of Wheat

Resistance to Septoria tritici blotch (STB) is an economically important trait in many wheat-breeding programs across the world. Several quantitative trait loci (QTL) for STB resistance were identified in wheat but due to the dynamic pathogen population it is necessary to continuously identify new resistance genes/QTL and determine the underlying resistance mechanism. In this work, we integrated QTL mapping and transcriptome profiling to identify candidate genes underlying QTL associated with STB resistance in bread wheat at the seedling stage. The results revealed four QTL on chromosomes 1BS, 1BL, 3AS and 3DL for STB resistance. Among these, two QTL on 2BL and 3DL were mapped for chlorosis, necrosis and pycnidia while the other two on 1BS and 3AS were associated with necrosis and pycnidia. Among the four identified QTL, genes were identified in three QTL (1BS, 2BL and 3DL). In total, 238 differentially expressed genes (DEGs) were localized in 1BS, 16 DEGs in 2BL and 80 DEGs in 3DL QTL region respectively. F-box protein, NBS-LRR disease resistance genes and receptor-like protein kinase were the most over-represented. The results emphasize the importance of integrating QTL and transcriptome analysis to accelerate the identification of key genes underlying the traits of interest.

QTL mapping of Fusarium head blight and Septoria tritici blotch in an elite hard red winter wheat population

The objective of this study was to map loci associated with Fusarium head blight (FHB) traits and Septoria tritici blotch (STB) and plant height in a “Maxine/FTHP Redeemer” winter wheat population. Evaluation of FHB and STB resistance was performed using spray inoculation and under natural infections in replicated trials across three environments in Ontario, Canada. Selected lines of this population were tested for DON level in 2013 (FHB epidemic year) at three locations in Ontario. For both diseases, the population showed a continuous distribution pattern. DArT markers were used to generate a genetic map, and quantitative trait loci (QTL) analysis was performed by evaluating 105 doubled-haploid lines. The most effective QTL associated with FHB resistance were identified on chromosomes 2D, 4A, and 4B, coinciding with plant height QTL, being an indicative of the presence of the dwarf alleles on Rht-B1 and Rht8 loci in the present population. The 2D QTL is probably due to Rht8 + Ppd-D1, because of the association of the 2D QTL with both plant height and heading date. STB QTL were on chromosomes 2D, 4B, and 7A. The lines with the genotype 2D+ (Rht8), combined with 4A− and 4B+ (Rht-B1) QTL, showed the best genetic combination to obtain short and early maturing plants with good resistance to FHB, low DON, and adequate resistance to STB. Under natural FHB conditions, in the 2013 epidemic year, the presence of 2D+ allele reduced the DON accumulation when compared with lines with 2D− allele. Both diseases were screened in the same inoculated nursery, but did not influence each other. Knowledge of the best genetic combination will allow the successful improvement of elite wheat cultivars.

An experimental approach for estimating the genomic selection advantage for Fusarium head blight and Septoria tritici blotch in winter wheat.

The genomic selection advantage for Fusarium head blight is promising but failed for Septoria tritici blotch. Selection of new breeding parents based on predictions must be treated with caution. Genomic selection (GS) is an approach that uses whole-genome marker data to estimate breeding values of untested genotypes and holds the potential to improve the genetic gain in breeding programs by shortening the breeding cycle and increasing the selection intensity. However, reported realized gain from genomic selection is limited to few experiments. In this study, a training population of 1120 winter wheat lines derived from 14 bi-parental families was genotyped with genome-wide single nucleotide polymorphism markers and phenotyped for Fusarium head blight (FHB) and Septoria tritici blotch (STB) severity, plant height and heading date. We used weighted ridge regression best linear unbiased prediction to calculate genomic estimated breeding values (GEBVs) of 2500 genotypes. Based on GEBVs, we selected the most resistant individuals as well as a random sample and tested them in a multi-location field trial. We computed moderate coefficients of correlation between observed and predicted trait values for FHB severity, plant height and heading date and achieved a genomic selection advantage of 10.62 percentage points for FHB resistance compared to the randomly chosen subpopulation. Genomic selection failed for the improvement of STB resistance with a genomic selection advantage of only 2.14 percentage points. Our results also indicate that the selection of new breeding parents based on GEBVs must be treated with caution. Taken together, the implementation of GS for FHB resistance, plant height and heading date seems to be promising. For traits with very strong genotype × environment variance, like STB resistance, GS appears to be still challenging.