Investigation of 2,4-Dihydroxylaryl-Substituted Heterocycles as Inhibitors of the Growth and Development of Biotrophic Fungal Pathogens Associated with the Most Common Cereal Diseases.

Climate change forces agriculture to face the rapidly growing virulence of biotrophic fungal pathogens, which in turn drives researchers to seek new ways of combatting or limiting the spread of diseases caused by the same. While the use of agrochemicals may be the most efficient strategy in this context, it is important to ensure that such chemicals are safe for the natural environment. Heterocyclic compounds have enormous biological potential. A series of heterocyclic scaffolds (1,3,4-thiadiazole, 1,3-thiazole, 1,2,4-triazole, benzothiazine, benzothiadiazine, and quinazoline) containing 2,4-dihydroxylaryl substituents were investigated for their ability to inhibit the growth and development of biotrophic fungal pathogens associated with several important cereal diseases. Of the 33 analysed compounds, 3 were identified as having high inhibitory potential against Blumeria and Puccinia fungi. The conducted research indicated that the analysed compounds can be used to reduce the incidence of fungal diseases in cereals; however, further thorough research is required to investigate their effects on plant-pathogen systems, including molecular studies to determine the exact mechanism of their activity.

Genetic mapping of the powdery mildew resistance gene Pm7 on oat chromosome 5D.

KEY MESSAGE: Three independent experiments with different genetic backgrounds mapped the resistance gene Pm7 in the oat genome to the distal part of the long arm of chromosome 5D. Resistance of oat to Blumeria graminis DC. f. sp. avenae is an important breeding goal in Central and Western Europe. In this study, the position of the effective and widely used resistance gene Pm7 in the oat genome was determined based on three independent experiments with different genetic backgrounds: genome-wide association mapping in a diverse set of inbred oat lines and binary phenotype mapping in two bi-parental populations. Powdery mildew resistance was assessed in the field as well as by detached leaf tests in the laboratory. Genotyping-by-sequencing was conducted to establish comprehensive genetic fingerprints for subsequent genetic mapping experiments. All three mapping approaches located the gene to the distal part of the long arm of chromosome 5D in the hexaploid oat genome sequences of OT3098 and 'Sang.' Markers from this region were homologous to a region of chromosome 2Ce of the C-genome species, Avena eriantha, the donor of Pm7, which appears to be the ancestral source of a translocated region on the hexaploid chromosome 5D.

Chemical Characteristics and Antioxidant Activity of Arctostaphylos uva-ursi L. Spreng. at the Southern Border of the Geographical Range of the Species in Europe.

The bearberry (Arctostaphylos uva-ursi L. Spreng.) is a source of herbal material-bearberry leaf (Uvae ursi folium), which is highly valued and sought by pharmaceutical and cosmetic industries. For many years, leaves of this plant have been used in traditional medicine as a diuretic, antimicrobial, and anti-inflammatory agent for various diseases of the urogenital tract. The bearberry has also been proposed as a natural antioxidant additive due to the high contents of phenolic compounds in its leaves. The study was focused on characterization of the basic phytochemical composition and antioxidant activity of extracts derived from bearberry leaves collected from plants located at the southern border of the geographical range of the species in Europe. The investigated herbal material is characterized by a different chemical profile compared to the chemical profiles of bearberry found in other parts of the continent. Bearberry extracts from plants growing in two different habitat types-heathlands and pine forests showed a wide range of variation, especially in the concentration of hyperoside, corilagin, and methylartutin and the total flavonoid contents. In addition to arbutin, bearberry can be a valuable source of phenolic compounds, which are mainly responsible for the antioxidant properties of extracts. The high content of phenols and high values of antioxidant parameters indicate a high potential of bearberry leaves to be used as a powerful natural source of antioxidants in herbal preparations. Therefore, the A. uva-ursi populations can be a source of plant material for pharmaceutical, cosmetic, and food industries.

Molecular identification and chromosomal localization of new powdery mildew resistance gene Pm11 in oat.

The appropriate selection of various traits in valuable plants is very important for modern plant breeding. Effective resistance to fungal diseases, such as powdery mildew, is an example of such a trait in oats. Marker-assisted selection is an important tool that reduces the time and cost of selection. The aims of the present study were the identification of dominant DArTseq markers associated with a new resistance gene, annotated as Pm11 and derived from Avena sterilis genotype CN113536, and the subsequent conversion of these markers into a PCR-based assay. Among the obtained 30,620 silicoDArT markers, 202 markers were highly associated with resistance in the analysed population. Of these, 71 were selected for potential conversion: 42 specific to resistant and 29 to susceptible individuals. Finally, 40 silicoDArT markers were suitable for primer design. From this pool, five markers, 3 for resistant and 2 for susceptible plants, were selected for product amplification in the expected groups. The developed method, based on 2 selection markers, provides certain identification of resistant and susceptible homozygotes. Also, the use of these markers allowed the determination of heterozygotes in the analysed population. Selected silicoDArT markers were also used for chromosomal localization of new resistance genes. Five out of 71 segregating silicoDArT markers for the Pm11 gene were found on the available consensus genetic map of oat. Five markers were placed on linkage groups corresponding to Mrg12 on the Avena sativa consensus map.

Resistance to Puccinia coronata f. sp. avenae in Avena magna, A. murphyi, and A. insularis.

Wild oat tetraploids of the section Pachycarpa have already been proven to be a rich source of useful genes but have largely been unexploited for Puccinia coronata resistance. In this study, accessions of Avena magna, A. murphyi, and A. insularis gathered from European and North American gene banks were evaluated at the seedling stage for crown rust reaction using the host-pathogen test and six highly diverse and virulent P. coronata isolates. Of the 92 Avena accessions analyzed, 58.7% were resistant to at least one crown rust race. In all, 37% of the tested accessions reacted nonuniformly, which indicated their heterogeneity. The highest level of resistance was observed in three of the accessions, one of which was verified by flow cytometry as being hexaploid and two of which were verified as being tetraploids. The infection profiles of 19 accessions corresponded to resistance determined by the genes Pc14, Pc39, Pc40, Pc48, Pc50, Pc54, Pc55, Pc61, Pc67, Pc68, Pc97, Pc101, or Pc104. The patterns of infection of the remaining resistant A. magna and A. murphyi accessions allowed us to postulate the presence of potentially novel crown rust resistance genes.

Avena sterilis L. Genotypes as a Potential Source of Resistance to Oat Powdery Mildew.

The aim of the present study was to identify Avena sterilis genotypes demonstrating a high level of resistance against oat powdery mildew, using host-pathogen tests. The study was conducted on 350 A. sterilis genotypes from different parts of the world. Six single-spore isolates of Blumeria graminis (DC.) f. sp. avenae, which demonstrated different levels of virulence to control lines and cultivars, were used in host-pathogen screening tests. To confirm the resistant response of selected genotypes, 13 other isolates were used. Reactions to the isolates were grouped into three classes: resistant, intermediate, and susceptible. Susceptible cultivars Sam and Fuchs were used as controls to estimate the degree of infection. The results of the screening test showed that 10 genotypes were classified as resistant. The second test based on 13 other isolates revealed that only four of the 10 genotypes were a valuable source of resistance against powdery mildew. The identified genotypes may be used in oat breeding programs to increase the level of resistance against powdery mildew. First, however, further studies aimed at identifying whether this resistance is conditioned by a single gene or combinations of different genes are required.

Resistant or Susceptible? How Central European Oat (A. sativa L.) Cultivars React to B. graminis f. sp. avenae Infection.

In accordance with the postulates of integrated plant protection, the use of cultivars with genetically determined resistance is one of the main strategies for preventing losses caused by fungal pathogens. The development of breeding programs aimed at increasing resistance to pathogens should be preceded by a characterization of the resistance of cultivars grown in a given area. This allows us to determine the number of genes used in breeding and their effectiveness. It also allows us to estimate the pressure that the pathogen may exert on varieties with specific resistance genes. The presented work aimed to determine the level of resistance of oat varieties currently cultivated in Central Europe and the number of effective powdery mildew resistance genes currently used in oat breeding programs. The research showed that out of 46 varieties, only 5 were resistant to powdery mildew. Analysis of the infection profiles allowed us to postulate the presence of the Pm7 gene in four of them. In the Merlin variety from the Czech Republic, it was not possible to determine which of the previously described genes determines resistance to powdery mildew. Due to the observed climate changes and the rapid adaptation of pathogens to new environmental conditions, it is crucial to introduce a wider pool of genes that determine the pathogen resistance of cultivars.

Chromosomal Location of Pm12-A Novel Powdery Mildew Resistance Gene from Avena sterilis.

Identification of new, effective disease resistance genes is a very important aspect of plant breeding. Also important is the precise location of individual loci and tagging them with DNA markers for marker assisted selection. The aim of the present study was identification of the molecular markers linked with Pm12, a new effective resistance gene to powdery mildew, and their location in the oat genome. The analysis was performed on 167 F2 individuals from a hybrid of Fuchs × CN67383, with the status of the locus in each individual verified by progeny test in F3. Segregation ratios confirmed the monogenic nature of resistance. Making use of the sequence data of DNA markers and the oat OT3098 v2 genome reference assembly, Pm12 is located on chromosome 7C. A comparison was also made with the reference consensus map, to which there are more reports of mapped genes to date. The mapping results suggest that Pm12 is located in the interval 103.8-111.7 cM on this map. No powdery mildew resistance locus has been identified in this region so far, suggesting that Avena sterilis CN67383 carries a novel locus offering effective resistance in oat breeding. The information included in the oat genome annotation allowed for the identification of candidate genes in the close region of the marker cluster for Pm12. This information may provide an interesting source of further analysis of the pathways of various genes in response to the stress of powdery mildew infection.

Virulence Structure and Genetic Diversity of Blumeria graminis f. sp. avenae from Different Regions of Europe.

The structure and dynamics of changes in pathogen populations can be analysed by assessing the level of virulence and genetic diversity. The aim of the present study was to determine the diversity of Blumeria graminis f. sp. avenae populations. Diversity and virulence of B. graminis f. sp. avenae was assessed based on 80 single-spore isolates collected in different European countries such as Poland (40 isolates), Germany (10), Finland (10), Czech Republic (10) and Ireland (10) using ISSR (Inter-Simple Sequence Repeats) and SCoT (Start Codon Targeted) markers. This work demonstrated differences in virulence of B. graminis f. sp. avenae isolates sampled from different countries. Molecular analysis showed that both systems were useful for assessing genetic diversity, but ISSR markers were superior and generated more polymorphic products, as well as higher PIC and RP values. UPMGA and PCoA divided the isolates into groups corresponding with their geographical origin. In conclusion, the low level of genetic differentiation of the analysed isolates has suggested that the evolution of B. graminis f. sp. Avenae population is slow, and thus the evolutionary potential of the pathogen is low. This work paves the way for future studies on B. graminis f. sp. Avenae population structure and dynamics based on genetic variability.

Genetic Similarity of Avena sativa L. Varieties as an Example of a Narrow Genetic Pool of Contemporary Cereal Species.

The assessment of the genetic diversity of cultivated varieties is a very important element of breeding programs. This allows the determination of the level of genetic differentiation of cultivated varieties, their genetic distinctiveness, and is also of great importance in the selection of parental components for crossbreeding. The aim of the present study was to determine the level of genetic diversity of oat varieties currently grown in Central Europe based on two marker systems: ISSR and SCoT. The research conducted showed that both these types of markers were suitable for conducting analyses relating to the assessment of genetic diversity. The calculated coefficients showed that the analyzed cultivars were characterized by a high genetic similarity. However, the UPGMA and PCoA analyses clearly indicated the distinctiveness of the breeding programs conducted in Central European countries. The high genetic similarity of the analyzed forms allow us to conclude that it is necessary to expand the genetic pool of oat varieties. Numerous studies show that landraces may be the donor of genetic variation.

Virulence Structure of Blumeria graminis f. sp. avenae Populations in Poland across 2014-2015.

The purpose of this study was to determine the virulence structure of oat powdery mildew (Blumeria graminis f. sp. avenae, Bga) populations in Poland collected in 2014 and 2015. Powdery mildew isolates were collected from 18 locations in Poland. In total, nine lines and cultivars of oat, with different mildew resistance genes, were used to assess virulence of 180 isolates. The results showed that a significant proportion of the Bga isolates found in Poland were virulent to differentials with Pm1, Pm3, Pm6, and Pm3 + Pm8 genes. In contrast Pm4, Pm5, Pm2, and Pm7 genes were classified as resistant to all pathogen isolates used in the experiment. Based on obtained results we can state that there are differences in virulence pattern and diversity parameters between sites and years, but clear trends are not deducible.

Molecular and physical characterization of grain hardness in European spring common wheat (Triticum aestivum L.).

Grain hardness is the single most important trait in determining technological properties and end-use quality of wheat product. This trait is controlled by two genes (Pina-D1 and Pinb-D1) at the Hardness (Ha) locus. Soft endosperm kernels are characterized by the presence of alleles 'a' in both genes (Pina-D1a and Pinb-D1a), while the medium hard and hard grain is the result of deletion in the Pina-D1 gene or single mutation of the Pinb-D1 gene. The aim of the current study was to determine the relationship between common wheat grain hardness and the presence of puroindoline genes. Eighty-one spring common wheat cultivars from Europe were analysed for grain hardness by SKCS (Single Kernel Characterization System) and Pin-D1 genes. The analysed genotypes were divided into three hardness classes: hard, medium and soft and they showed four allelic combinations in Pin-D1 genes. The SKCS results showed that hard wheat was the major type in European cultivars, whereas molecular analysis showed differential allelic combinations of puroindoline genes among these classes. The conducted analyses suggest that another major gene or other factors were influencing kernel texture. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02897-3.

Is Every Wild Species a Rich Source of Disease Resistance? Avena fatua L.-Potential Donor of Resistance to Powdery Mildew.

Identifying effective sources of disease resistance is an important aspect of an effective plant protection strategy. Wild species related to cultivars constitute a rich reservoir of resistance genes. Studies conducted in oat have shown that wild species are donors of resistance genes to crown and stem rust, powdery mildew or fusarium head blight. The aim of the present study was to prove whether A. fatua could be a source of effective resistance genes to powdery mildew. This species is widespread all over the world due to its very good adaptability and can be regarded as a potential source of resistance to fungal diseases, including powdery mildew. The conducted research has shown that A. fatua is a species with a low level of resistance to powdery mildew when compared to other wild species of the genus Avena L. A total of 251 accessions were evaluated, and only 23 were identified as resistant to the individual isolates used in the host-pathogen tests. It follows that resistance to powdery mildew is not common among wild Avena species, and its good environmental adaptation is not associated to resistance to powdery mildew.