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.

Key message Wheat blast resistance in Caninde#1 is controlled by a major QTL on 2NS/2AS translocation and multiple minor QTL in an additive mode. Abstract Wheat blast (WB) is a devastating disease in South America, and it recently also emerged in Bangladesh. Host resistance to WB has relied heavily on the 2NS/2AS translocation, but the responsible QTL has not been mapped and its phenotypic effects in different environments have not been reported. In the current study, a recombinant inbred line population with 298 progenies was generated, with the female and male parents being Caninde#1 (with 2NS) and Alondra (without 2NS), respectively. Phenotyping was carried out in two locations in Bolivia, namely Quirusillas and Okinawa, and one location in Bangladesh, Jashore, with two sowing dates in each of the two cropping seasons in each location, during the years 2017-2019. Genotyping was performed with the DArTseq® technology along with five previously reported STS markers in the 2NS region. QTL mapping identified a major and consistent QTL on 2NS/2AS region, explaining between 22.4 and 50.1% of the phenotypic variation in different environments. Additional QTL were detected on chromosomes 1AS, 2BL, 3AL, 4BS, 4DL and 7BS, all additive to the 2NS QTL and showing phenotypic effects less than 10%. Two codominant STS markers, WGGB156 and WGGB159, were linked proximally to the 2NS/2AS QTL with a genetic distance of 0.9 cM, being potentially useful in marker-assisted selection.

Key Message Using disease bioassays and transcriptomic analysis we show that intact SA-signalling is required for potato defences against the necrotrophic fungal pathogen Alternaria solani. Abstract Early blight, caused by the necrotrophic fungus Alternaria solani, is an increasing problem in potato cultivation. Studies of the molecular components defining defence responses to A. solani in potato are limited. Here, we investigate plant defence signalling with a focus on salicylic acid (SA) and jasmonic acid (JA) pathways in response to A. solani. Our bioassays revealed that SA is necessary to restrict pathogen growth and early blight symptom development in both potato foliage and tubers. This result is in contrast to the documented minimal role of SA in resistance of Arabidopsis thaliana against necrotrophic pathogens. We also present transcriptomic analysis with 36 arrays of A. solani inoculated SA-deficient, JA-insensitive, and wild type plant lines. A greater number of genes are differentially expressed in the SA-deficient mutant plant line compared to the wild type and JA-insensitive line. In wild type plants, genes encoding metal ion transporters, such as copper, iron and zinc transporters were upregulated and transferase-encoding genes, for example UDP-glucoronosyltransferase and Serine-glyoxylate transferase, were downregulated. The SA-deficient plants show upregulation of genes enriched in GO terms related to oxidoreductase activity, respiratory chain and other mitochondrial-related processes. Pathogenesis-related genes, such as genes encoding chitinases and PR1, are upregulated in both the SA-deficient and wild type plants, but not in the JA-insensitive mutants. The combination of our bioassays and the transcriptomic analysis indicate that intact SA signalling, and not JA signalling, is required for potato defences against the necrotrophic pathogen A. solani.

BackgroundEarly blight, caused by the fungus Alternaria solani, occurs on potato mainly in the south-eastern part of Sweden, but also in other parts of the country. The aim of this study was to investigate the genetic diversity of A. solani populations from different potato growing regions in south-eastern Sweden using AFLP marker analysis. In addition, the cultured isolates were examined for substitutions in the gene encoding cytochrome b, associated with loss of sensitivity against QoI fungicides.ResultsNei's gene diversity index for the Swedish populations of A. solani revealed a gene diversity of up to 0.20. Also genetic differentiation was observed among populations of A. solani from different locations in south-eastern Sweden. The mitochondrial genotype of the isolates of A. solani was determined and both known genotypes, GI (genotype 1) and GII (genotype 2), were found among the isolates. The occurrence of the F129L substitution associated with a loss of sensitivity to strobilurins was confirmed among the GII isolates. In vitro conidial germination tests verified that isolates containing the F129L substitution had reduced sensitivity to azoxystrobin and, at a lower extent, to pyraclostrobin.ConclusionsGenetic diversity was relatively high among isolates of A. solani in south-eastern part of Sweden. F129L substitutions, leading to reduced sensitivity to strobilurins, have been established in field populations, which may have implications for the future efficacy of QoI fungicides.

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Early blight, caused by the fungus Alternaria solani, is a common foliar disease in potato. Quinone outside inhibitor (QoIs) fungicides have commonly been used against A. solani. To avoid or delay development of fungicide resistance it is recommended to alternate or combine fungicides with different modes of action. Therefore, we compared two different fungicide programs against early blight in field trials and studied within season changes in the pathogen population. An untreated control was compared with treatments using azoxystrobin alone and with a program involving difenoconazole followed by boscalid and pyraclostrobin combined. Isolates of A. solani were collected during the growing season and changes in the population structure was investigated. We also screened for the amino acid substitution in the cytochrome b gene and investigated changes in sensitivity to azoxystrobin. Treatment with azoxystrobin alone did not improve disease control in 2014 when the disease pressure was high. However, lower severity of the disease was observed after combined use of difenoconazole, boscalid and pyraclostrobin. The efficacy of both fungicide treatments were similar during the field trial in 2017. Two mitochondrial genotypes (GI and GII) were found among isolates, where all isolates, except two, were GII. All GII isolates had the F129 L substitution while the two GI isolates were wild type. Population structure analysis and principal component analysis (PCA) of amplified fragment length polymorphisms (AFLP) data revealed within season changes in the A. solani populations in response to fungicide application. Isolates with the F129 L substitution had reduced sensitivity to azoxystrobin in vitro and their sensitivity tended to decrease with time.

Early blight of potato, caused by Alternaria solani, is an economically important foliar disease in most potato-growing regions. Growing cultivars with higher levels of resistance to early blight can reduce tuber yield losses and the need for fungicide applications. In this research, a bi-parental tetraploid potato population, segregating for resistance to early blight in leaves and tubers, was characterized to identify novel quantitative trait loci (QTL) associated with foliar and tuber early blight resistance. Assessment of the disease resistance in the foliage was performed by field evaluation and in tuber under controlled conditions. Results from this study revealed significant differences (P < 0.001) in resistance to A. solani among potato clones both in the leaves and in tubers. There was no statistically significant correlation (r = 0.06, P = 0.35) between the resistance scores from leaves and tubers. Several clones exhibited; however, high levels of resistance both in leaves and tubers and are; thus, promising candidates for breeding for early blight resistance. Linkage mapping revealed several QTL for early blight affecting both foliage and tubers. QTL associated with disease resistance in the tuber were found on chromosomes 1, 2, 3, 4, 8, 11 and 12. QTL associated with disease resistance in foliage were also examined for independence from defoliation, and independent QTL were; thus, found on chromosomes 5 and 11.

Plant phenotyping by imaging allows automated analysis of plants for various morphological and physiological traits. In this work, we developed a low-cost RGB imaging phenotyping lab (LCP lab) for low-throughput imaging and analysis using affordable imaging equipment and freely available software. LCP lab comprising RGB imaging and analysis pipeline is set up and demonstrated with early vigour analysis in wheat. Using this lab, a few hundred pots can be photographed in a day and the pots are tracked with QR codes. The software pipeline for both imaging and analysis is built from freely available software. The LCP lab was evaluated for early vigour analysis of five wheat cultivars. A high coefficient of determination (R2 0.94) was obtained between the dry weight and the projected leaf area of 20-day-old wheat plants and R2 of 0.9 for the relative growth rate between 10 and 20 days of plant growth. Detailed description for setting up such a lab is provided together with custom scripts built for imaging and analysis. The LCP lab is an affordable alternative for analysis of cereal crops when access to a high-throughput phenotyping facility is unavailable or when the experiments require growing plants in highly controlled climate chambers. The protocols described in this work are useful for building affordable imaging system for small-scale research projects and for education.