Short Communication. Virulence of barley leaf rust in the South of Russia in 2017-2019

Aim of study: To analyze the structure of Puccinia hordei populations by virulence in southern Russia during 2017-2019. Area of study: South of Russia, the leading Russian region for barley production where barley leaf rust is an important foliar disease. Material and methods: Uredinial samples of P. hordei were collected at the production sites of winter barley in the south of Russia. Single uredinial isolates (total 95) were tested for virulence with 17 differentials with Rph resistance genes. Main results: No isolates found virulent to the host line with the Rph13 gene. There was a decrease in the number of fungal isolates virulent to the host lines with Rph5 and Rph7 genes. In 2017 and 2019, isolates containing a large number of virulence alleles (from 11 to 15) prevailed. In 2018, isolates with low (1-5) and medium (6-10) frequency of virulent alleles prevailed, as well as avirulent isolates. The values of the Nei index via diversity showed high similarity of the pathogen populations in 2017-2018 (N = 0.05) and minor differences in 2017-2019 and 2018-2019 (N = 0.13 and 0.16, respectively). The greatest frequency of virulence alleles in accordance with the Nei (Hs) index was noted for the 2018 population (Hs = 0.36). For the 2017 and 2019 populations, this indicator was average (Hs = 0.29 and 0.20, respectively). Research highlights: Analysis of genetics of the P. hopdei population is important for the strategy of varietal distribution in the region and development of rust-resistant cultivars.

Under Attack: Fungal Landscape of Spring Barley From Seedling Emergence to Harvest During a Severe Puccinia Hordei Epidemic

All plant tissues from leaves, stems and roots are hosting a wide diversity of fungal species. Our understanding of the assembly of this diversity of fungi during the plant growth cycle is limited. Here, we characterized the mycobiome of three spring barley cultivars at weekly intervals from seedling emergence to senescence and seed maturity. A notable proportion of members of the fungal communities were shared among different plant organs, but community dynamics were tissue specific. A severe attack of Puccinia hordei occurring during the vegetative stage had profound effects on the mycobiome, and P. hordei biomass displaced that of other taxa. Plant tissue type was the most important factor determining the mycobiome, but also plant age was contributing significantly. Using a random forest model, we found that specific members of the mycobiome were responding differently to plant age. A co-occurrence network analysis revealed complex interactions among fungal OTUs, and network connectivity was changing as per plant growth stage and plant tissue type. This study contributes to the understanding of assembly of fungal communities in cereals by providing a detailed atlas of fungal communities associated with barley. This knowledge will be vital for microbiome assisted plant health management and our study will serve as an important baseline for future efforts to harness microbiota in cereal health.

Mining and predictive characterization of resistance to leaf rust (Puccinia hordei Otth) using two subsets of barley genetic resources

Sustainable barley (Hordeum vulgare L.) production will require access to diverse ex-situ conserved collections to develop varieties with high yields and capable of overcoming the challenges imposed by major abiotic and biotic stresses. This study aimed at searching efficient approaches for the identification of new sources of resistance to barley leaf rust (Puccinia hordei Otth). Two subsets, Generation Challenge Program Reference set (GCP) with 188 accessions and leaf rust subset constructed using the filtering approach of the Focused Identification of Germplasm Strategy (FIGS) with 86 accessions, were evaluated for the seedling as well as the adult plant stage resistance (APR) using two barley leaf rust (LR) isolates (ISO-SAT and ISO-MRC) and in four environments in Morocco, respectively. Both subsets yielded a high percent of accessions with a moderately resistant (MR) reaction to the two LR isolates at the seedling stage. For APR, more than 50% of the accessions showed resistant reactions in SAT2018 and GCH2018, while this rate was less than 20% in SAT2017 and SAT2019. Statistical analysis using chi-square test of independence revealed the dependency of LR reaction on subsets at the seedling (ISO-MRC), as well as at the APR (SAT2017 and SAT2018) stage. At seedling stage, the test of goodness of fit showed that GCP subset yielded higher percentages of resistant accessions than FIGS-LR in case of ISO-MRC isolate but the two subsets did not differ for ISO-SAT. At APR, FIGS approach performed better than GCP in yielding higher percentages of accessions in case of SAT2017 and SAT2018. Although some of the tested machine learning models had moderate to high accuracies, none of them was able to find a strong and significant relationship between the reaction to LR and the environmental conditions showing the needs for more fine tuning of approaches for efficient mining of genetic resources using machine learning.

The barley leaf rust resistance gene Rph3 encodes a putative executor protein

Host resistance is considered the most effective means to control plant diseases; however, individually deployed resistance genes are often rapidly overcome by pathogen adaptation. Combining multiple effective resistance genes is the optimal approach to durable resistance, but the lack of functional markers for resistance genes has hampered implementation. Leaf rust, caused by Puccinia hordei, is an economically significant disease of barley, but only a few major Resistance genes to P. hordei (Rph) have been cloned. In this study, gene Rph3 was isolated by positional cloning and confirmed by mutational analysis and transgenic complementation. The Rph3 gene, which originated from wild barley and was first introgressed into cultivated Egyptian germplasm, encodes a unique transmembrane resistance protein that differs from all known plant disease resistance proteins at the amino acid sequence level. Genetic profiles of diverse accessions indicated limited genetic diversity in Rph3 in domesticated germplasm, and higher diversity in wild barley from the Eastern Mediterranean region. Expression profiling using P. hordei isolates with contrasting pathogenicity for the Rph3 host locus showed that the Rph3 gene was expressed only in interactions with Rph3-avirulent isolates, a phenomenon also observed for transcription activator-like effector-dependent genes known as executors conferring resistance to Xanthomonas spp. Like the known transmembrane executors such as Bs3 and Xa7 heterologous expression of Rph3 in N. benthamiana induced a cell death response. Given that Rph3 shares several features with executor genes, it seems likely that P. hordei contains effectors similar to the transcription activator-like effectors that target host executor genes. The isolation of Rph3 highlights convergent evolutionary processes in diverse plant-pathogen interaction systems, where similar defence mechanisms evolved independently in monocots and dicots and provide evidence for executor genes in the Triticeae tribe.

Mining and predictive characterization for resistance to leaf rust (Puccinia hordei Otth) using two subsets of barley genetic resources

Sustainable barley production will require access to diverse ex-situ conserved collections to develop varieties with high yields and capable of overcoming the challenges imposed by major abiotic and biotic stresses. This study aimed at searching efficient approaches for the identification of new sources of resistance to barley leaf rust (LR). Two subsets, Generation Challenge Program Reference set (GCP) with 190 accessions and leaf rust subset constructed using the filtering approach of the Focused Identification of Germplasm Strategy (FIGS) with 100 accessions, were evaluated for the seedling as well as the adult plant stage resistance (APR) using two LR isolates (ISO-SAT and ISO-MRC) and in four environments in Morocco, respectively. Both subsets yielded a high percent of accessions with a moderately resistant (MR) reaction to the two LR isolates at the seedling stage. For APR, more than 50% of the accessions showed resistant reactions in SAT2018 and GCH2018, while this rate was less than 20% in SAT2017 and SAT2019. Statistical analysis using chi-square test of independence revealed the dependency of LR reaction on subsets at the seedling (ISO-MRC), as well as at the APR (SAT2017 and SAT2018) stage. Furthermore, the test of goodness of fit showed that FIGS_LR yielded higher percentages of resistant accessions than GCP subset in case of ISO-MRC at the seedling stage, and in case of SAT2017 and SAT2018 at APR stage. Although some of the tested machine learning models had moderate to high accuracies, none of them was able to find a strong and significant relationship between the reaction to LR and the environmental conditions showing the needs for more fine tuning of approaches for efficient mining of genetic resources using machine learning.

Leaf rust resistance in barley accessions from Ethiopia

Currently, the problem of broadening the diversity of cultivated barley varieties based on the effective genes for resistance to the leaf rust causal agent Puccinia hordei has become relevant. In 2018–2019 the resistance of 925 barley accessions from Ethiopia to the northwestern (St. Petersburg, Pushkin) pathogen population was evaluated. The accessions were sown at a later date, favoring to severe plant damage. The resistance was assessed during the heading period and over the milk ripening stage using a point scale. The results of the experiments have indicated a low diversity of Ethiopian barley according to the studied trait. Only 4 accessions have been resistant to P. hordei, and 3 breeding lines (k-30810, k-30811 and k-30812) have been protected by the previously identified Rph7 gene, which efficiency has been decreasing in recent years. The accession k-21919 has possessed a high level of resistance to the pathogen (i.e. lack of damage symptoms) and therefore it can be recommended for utilization in barley breeding.

Validating molecular markers for barley leaf rust resistance genes Rph20 and Rph24

Improving resistance to barley leaf rust (caused by Puccinia hordei) is an important breeding objective in most barley growing regions worldwide. The development and subsequent utilization of high-throughput polymerase chain reaction (PCR) based co-dominant molecular markers remains an effective approach to select genotypes with multiple effective resistance genes, permitting efficient gene deployment and stewardship. The genes Rph20 and Rph24, which confer widely effective adult plant resistance (APR) to leaf rust, are common in European and Australian barley germplasm (often in combination), and act interactively to confer high levels of resistance. Here we report on the development and validation of co-dominant insertion-deletion (indel) based PCR markers that are highly predictive for the resistance alleles Rph20.ai and Rph24.an (both referred to as Rph20 and Rph24).