Search Results (25)

Soilborne diseases cause losses of 45–70% in faba bean in Ethiopia. Studies were undertaken to define soilborne pathogens and their complexes in Ethiopia. First, the severity of root rot was assessed in 150 field sites across seven Ethiopian regions. Soil samples were collected, and the DNA of 29 pests and pathogens was quantified using a commercial quantitative PCR (qPCR) soil testing service. There was a very high incidence rate of Macrophomina phaseolina, as well as Pythium clades F and I. The other detected species in order of incidence included Fusarium redolens, Rhizoctonia solani, Aphanomyces euteiches, Phytophthora megasperma, Sclerotinia sclerotiorum and S. minor, and Verticillium dahliae, as well as low levels of Thielaviopsis basicola. Five anastomosis groups (AG) of R. solani, namely AG2.1, AG2.2, AG3, AG4, and AG5, were detected, of which AG2.2 and AG4 were most prevalent. We believe this is the first report of occurrence for Ethiopia of A. euteiches, Ph. megasperma, T. basicola, and the five AGs for R. solani. There were very high incidence rates of the foliar pathogens Botrytis cinerea, B. fabae, Didymella pinodes, and Phoma pinodella and of the nematode Pratylenchus thornei, followed by P. neglectus and P. penetrans. The root rot severity and distribution varied significantly across regions, as well as with soil types, soil pH, and soil drainage. Subsequently, metabarcoding of the soil DNA was undertaken using three primer pairs targeting fungi (ITS2), Fusarium species (TEF1 α), and Oomycetes (ITS1Oo). The ITS2 and TEF1α primers emphasized F. oxysporum as the most abundant soilborne fungal pathogen and highlighted F. ananatum, F. brachygibbosum, F. brevicaudatum, F. clavum, F. flagelliforme, F. keratoplasticum, F. napiforme, F. nelsonii, F. neocosmosporiellum, F. torulosum, and F. vanettenii as first reports of occurrence for Ethiopia. The ITS1Oo primer confirmed Pythium spp. as the most prevalent of all Oomycetes. Full article

This paper provides data on non-native fungus-like organisms (Oomycota, kingdom Stramenopila), their diversity, and distribution in Lithuania and is an addition to a previously published checklist of alien true fungi (Chytridiomycota, Ascomycota, and Basidiomycota; kingdom Fungi). All available published data were summarized, and the analysis showed that approximately 25% of recorded Lithuania stramenopilous fungus-like organism species are alien. The compiled list represented herein includes 54 species of alien terrestrial and aquatic stramenopilous fungus-like organisms (Oomycota). Most alien Oomycota are obligate pathogens of terrestrial plants. Aquatic organisms are represented by one alien dangerous crayfish pathogen, Aphanomyces astaci, which is currently noted as extinct in Lithuania. The taxonomy of the listed alien species of Oomycota has been revised, and the species names have been adjusted to reflect current nomenclature changes. The taxonomical and ecological analysis demonstrated the prevalence of terrestrial parasitic Peronosporales reported in Lithuania Oomycota and the highest diversity of alien species belonging to the Peronospora genus. The chronology of appearance, distribution within the country, and current status of these non-native organisms were assessed, and some of the most economically or ecologically important harmful invasive species were discussed. Full article

We adopted a morphometric approach to provide statistical support for the description of two different morphotypes (I, reproductive, II, non-reproductive) firstly observed in Procambarus clarkii specimens caught in a population from Sardinia Island (western Mediterranean). The morphometric study was preceded by molecular taxonomic identification using the mitochondrial Cytochrome C Oxidase subunit I (COI) gene. The presence or absence of the pathogen Aphanomyces astaci, responsible for the plague, was also investigated using the ribosomal Internal Transcribed Spacer (ITS) marker. The estimation of the size at onset of maturity (SOM) was further performed in males. Finally, the population structure was examined. Our results confirmed the entirety of the individuals as P. clarkii, and the absence of the pathogen A. astaci. Morphological analysis revealed a significant correlation between chela and carapace lengths, while intra- and inter-sexual dimorphism in the chela length confirmed the occurrence of the two morphotypes. The SOM in males was established between 35.0 and 37.1 mm carapace length. Finally, the size-population structure showed a Gaussian distribution. The recognition of the two distinct morphotypes in the crayfish could be useful as an additional tool not only to identify the reproductive period of this invasive crustacean but also to enrich the guidelines for the correct taxonomic identification of the species. Full article

Aphanomyces root rot (ARR), caused by Aphanomyces euteiches, is one of the most devastating diseases that affect the production of peas. Several control strategies such as crop rotation, biocontrol, and fungicides have been proposed, but none provides a complete solution. Therefore, the deployment of resistant cultivars is fundamental. ARR resistance breeding is hampered by the moderate levels of resistance identified so far. The available screening protocols require post-inoculation root assessment, which is destructive, time-consuming, and tedious. In an attempt to address these limitations, we developed a non-destructive screening protocol based on foliar symptoms and used it to identify new sources of resistance in a Pisum spp. germplasm collection. Accessions were root inoculated separately with two A. euteiches isolates, and leaf symptoms were assessed at 5, 10, 14, 17, and 20 days after inoculation (DAI). Although the majority of accessions exhibited high levels of susceptibility, thirty of them exhibited moderate resistance. These thirty accessions were selected for a second experiment, in which they were inoculated with both A. euteiches isolates at two inoculum doses. The objective of this second trial was to confirm the resistance of these accessions by evaluating root and biomass loss, as well as foliar symptoms, and to compare root and foliar evaluations. As a result, a high correlation (R² = 0.75) between foliar and root evaluations was observed, validating the foliar evaluation method. Notably, accessions from P.s. subsp. humile exhibited the lowest symptomatology across all evaluation methods, representing valuable genetic resources for breeding programs aimed at developing pea varieties resistant to ARR. Full article

Crayfish plague is a devastating disease of European freshwater crayfish and is caused by the oomycete Aphanomyces astaci (Ap. astaci), believed to have been introduced to Europe around 1860. All European species of freshwater crayfish are susceptible to the disease, including the white-clawed crayfish Austropotamobius pallipes. Ap. astaci is primarily spread by North American crayfish species and can also disperse rapidly through contaminated wet gear moved between water bodies. This spread, coupled with competition from non-indigenous crayfish, has drastically reduced and fragmented native crayfish populations across Europe. Remarkably, the island of Ireland remained free from the crayfish plague pathogen for over 100 years, providing a refuge for A. pallipes. However, this changed in 1987 when a mass mortality event was linked to the pathogen, marking its introduction to the region. Fortunately, crayfish plague was not detected again in Ireland until 2015 when a molecular analysis linked a mass mortality event in the Erne catchment to Ap. astaci. Since then, the pathogen has appeared across the island. Between 2015 and 2023, Ap. astaci was detected in 18 water catchments, revealing multiple genotypes. Intriguingly, the pathogen in Ireland is present without its natural host species. The uneven distribution of various genetic lineages strongly suggests the human-mediated transport of zoospores via contaminated water equipment as a primary cause of spread. This review details the timeline of these events, Ap. astaci’s introduction into Ireland, and its rapid spread. As well, this review references the genotypes that have been determined, and discusses the issue of non-indigenous crayfish species in Ireland and management efforts. Full article

In recent years, a new root rot disease in barley, which is caused by an Aphanomyces species, was found in field surveys in Southern Sweden and Denmark. Its symptoms occurred at the early tillering stage, around the BBCH 21 growth stage, and included the yellowing of leaves, brown coleoptiles, and the discolouration of roots. Prolonged soil wetness after rainfall favoured disease development, which sometimes advanced the yellowing patches to entire fields, resulting in lower yields. Oospores were found in the fine roots of diseased plants, and Aphanomyces isolates were obtained from these roots, as well as from the roots of barley plants grown in the greenhouse in soil samples from infected fields. Based on morphological analysis, we found that the new isolates were similar to those already obtained from barley and spinach roots in the 1990s in the same growing area. The morphological and molecular analyses performed in this study clearly separated and distinguished these barley isolates from other known Aphanomyces, and hereby Aphanomyces macrosporus sp. nov. is proposed as a new plant pathogenic species. It has larger oogonia and oospores than A. euteiches, A. cochlioides, and A. cladogamus, with one up to eight diclinous antheridia per oogonium. The phylogenetic analysis of the ITS rDNA region sequences grouped these new Aphanomyces isolates in a monophyletic clade, which was clearly distinguished from other plant pathogenic Aphanomyces species. The further pathogenicity of A. macrosporus on other plants is currently under investigation, but it is clear that it can at least infect barley, spinach, and sugar beet, indicating a wide host range for this species. The widespread presence and presumably broad host range of this new pathogenic Aphanomyces species must be considered in crop rotations. Full article

Small molecules as well as peptide-based therapeutic approaches have attracted global interest due to their lower or no toxicity in nature, and their potential in addressing several health complications including immune diseases, cardiovascular diseases, metabolic disorders, osteoporosis and cancer. This study proposed a peptide, GE18 of subtilisin-like peptidase from the virulence factor of aquatic pathogenic fungus Aphanomyces invadans, which elicits anti-cancer and anti-microbial activities. To understand the potential GE18 peptide-induced biological effects, an in silico analysis, in vitro (L6 cells) and in vivo toxicity assays (using zebrafish embryo), in vitro anti-cancer assays and anti-microbial assays were performed. The outcomes of the in silico analyses demonstrated that the GE18 peptide has potent anti-cancer and anti-microbial activities. GE18 is non-toxic to in vitro non-cancerous cells and in vivo zebrafish larvae. However, the peptide showed significant anti-cancer properties against MCF-7 cells with an IC50 value of 35.34 µM, at 24 h. Besides the anti-proliferative effect on cancer cells, the peptide exposure does promote the ROS concentration, mitochondrial membrane potential and the subsequent upregulation of anti-cancer genes. On the other hand, GE18 elicits significant anti-microbial activity against P. aeruginosa, wherein GE18 significantly inhibits bacterial biofilm formation. Since the peptide has positively charged amino acid residues, it targets the cell membrane, as is evident in the FESEM analysis. Based on these outcomes, it is possible that the GE18 peptide is a significant anti-cancer and anti-microbial molecule. Full article

In endangered crayfish conservation projects, it is paramount to map the distribution of the causative agent of crayfish plague, Aphanomyces astaci, in native populations. Considering the inapplicability of the destructive cuticular sampling protocol for monitoring endangered populations, we explored the use of non-invasive sampling techniques to detect this pathogen with molecular assays. In the present study, we exploited environmental DNA (testing increasing water volumes combined with different filter porosities) and cuticular swabs to collect A. astaci DNA. In addition, we evaluated the impact of the storage method on DNA preservation during field activities. After the first evaluations performed on both highly infected Austropotamobius pallipes and carrier Procambarus clarkii specimens in laboratory conditions, these sampling techniques were applied to wild populations of white-clawed crayfish. Our findings highlight better results with the filtration of 5 L of water with filters of 2.7 µm porosity for eDNA analysis and demonstrate that cuticular swabbing is equally effective as the World Organisation of Animal Health’s protocol. Storage in absolute ethanol proved to be the best solution to preserve swabs and filter samples for up to a week at room temperature. In conclusion, we suggest an integration of both sampling methods when monitoring A. astaci for conservation purposes. Full article

Aphanomyces invadans is an aquatic oomycete pathogen and the causative agent of epizootic ulcerative syndrome (EUS) in fresh and brackish water fish, which is responsible for severe mortalities and economic losses in aquaculture. Therefore, there is an urgent need to develop anti-infective strategies to control EUS. An Oomycetes, a fungus-like eukaryotic microorganism, and a susceptible species, i.e., Heteropneustes fossilis, are used to establish whether an Eclipta alba leaf extract is effective against the EUS-causing A. invadans. We found that treatment with methanolic leaf extract, at concentrations between 50–100 ppm (T4–T6), protects the H. fossilis fingerlings against A. invadans infection. These optimum concentrations induced anti-stress and antioxidative response in fish, marked by a significant decrease in cortisol and elevated levels of superoxide dismutase (SOD) and catalase (CAT) levels in treated animals, as compared with the controls. We further demonstrated that the A. invadans-protective effect of methanolic leaf extract was caused by its immunomodulatory effect and is linked to the enhanced survival of fingerlings. The analysis of non-specific and specific immune factors confirms that methanolic leaf extract-induced HSP70, HSP90 and IgM levels mediate the survival of H. fossilis fingerlings against A. invadans infection. Taken together, our study provides evidence that the generation of anti-stress and antioxidative responses, as well as humoral immunity, may play a role in protecting H. fossilis fingerlings against A. invadans infection. It is probable that E. alba methanolic leaf extract treatment might become part of a holistic strategy to control EUS in fish species. Full article

In this study, the anti-cancer and anti-inflammatory activities of PS14, a short peptide derived from the cellulase binding domain of pathogenic fungus, Aphanomyces invadans, have been evaluated, in vitro and in vivo. Bioinformatics analysis of PS14 revealed the physicochemical properties and the web-based predictions, which indicate that PS14 is non-toxic, and it has the potential to elicit anti-cancer and anti-inflammatory activities. These in silico results were experimentally validated through in vitro (L6 or Hep-2 cells) and in vivo (zebrafish embryo or larvae) models. Experimental results showed that PS14 is non-toxic in L6 cells and the zebrafish embryo, and it elicits an antitumor effect Hep-2 cells and zebrafish embryos. Anticancer activity assays, in terms of MTT, trypan blue and LDH assays, showed a dose-dependent inhibitory effect on cell proliferation. Moreover, in the epithelial cancer cells and zebrafish embryos, the peptide challenge (i) caused significant changes in the cytomorphology and induced apoptosis; (ii) triggered ROS generation; and (iii) showed a significant up-regulation of anti-cancer genes including BAX, Caspase 3, Caspase 9 and down-regulation of Bcl-2, in vitro. The anti-inflammatory activity of PS14 was observed in the cell-free in vitro assays for the inhibition of proteinase and lipoxygenase, and heat-induced hemolysis and hypotonicity-induced hemolysis. Together, this study has identified that PS14 has anti-cancer and anti-inflammatory activities, while being non-toxic, in vitro and in vivo. Future experiments can focus on the clinical or pharmacodynamics aspects of PS14. Full article

Aphanomyces root rot (ARR) is a devastating disease that affects the production of pea. The plants are prone to infection at any growth stage, and there are no chemical or cultural controls. Thus, the development of resistant pea cultivars is important. Phenomics technologies to support the selection of resistant cultivars through phenotyping can be valuable. One such approach is to couple imaging technologies with deep learning algorithms that are considered efficient for the assessment of disease resistance across a large number of plant genotypes. In this study, the resistance to ARR was evaluated through a CNN-based assessment of pea root images. The proposed model, DeepARRNet, was designed to classify the pea root images into three classes based on ARR severity scores, namely, resistant, intermediate, and susceptible classes. The dataset consisted of 1581 pea root images with a skewed distribution. Hence, three effective data-balancing techniques were identified to solve the prevalent problem of unbalanced datasets. Random oversampling with image transformations, generative adversarial network (GAN)-based image synthesis, and loss function with class-weighted ratio were implemented during the training process. The result indicated that the classification F1-score was 0.92 ± 0.03 when GAN-synthesized images were added, 0.91 ± 0.04 for random resampling, and 0.88 ± 0.05 when class-weighted loss function was implemented, which was higher than when an unbalanced dataset without these techniques were used (0.83 ± 0.03). The systematic approaches evaluated in this study can be applied to other image-based phenotyping datasets, which can aid the development of deep-learning models with improved performance. Full article

Aphanomyces root rot, caused by Aphanomyces euteiches, causes severe yield loss in field pea (Pisum sativum). The identification of a pea germplasm resistant to this disease is an important breeding objective. Polygenetic resistance has been reported in the field pea cultivar ‘00-2067’. To facilitate marker-assisted selection (MAS), bulked segregant RNA-seq (BSR-seq) analysis was conducted using an F₈ RIL population derived from the cross of ‘Carman’ × ‘00-2067’. Root rot development was assessed under controlled conditions in replicated experiments. Resistant (R) and susceptible (S) bulks were constructed based on the root rot severity in a greenhouse study. The BSR-seq analysis of the R bulks generated 44,595,510~51,658,688 reads, of which the aligned sequences were linked to 44,757 genes in a reference genome. In total, 2356 differentially expressed genes were identified, of which 44 were used for gene annotation, including defense-related pathways (jasmonate, ethylene and salicylate) and the GO biological process. A total of 344.1 K SNPs were identified between the R and S bulks, of which 395 variants were located in 31 candidate genes. The identification of novel genes associated with partial resistance to Aphanomyces root rot in field pea by BSR-seq may facilitate efforts to improve management of this important disease. Full article

Antibiosis has been proposed to contribute to the beneficial bacteria-mediated biocontrol against pea Aphanomyces root rot caused by the oomycete pathogen Aphanomyces euteiches. However, the antibiotics required for disease suppression remain unknown. In this study, we found that the wild type strains of Pseudomonas protegens Pf-5 and Pseudomonas fluorescens 2P24, but not their mutants that lack 2,4-diacetylphloroglucinol, strongly inhibited A. euteiches on culture plates. Purified 2,4-diacetylphloroglucinol compound caused extensive hyphal branching and stunted hyphal growth of A. euteiches. Using a GFP-based transcriptional reporter assay, we found that expression of the 2,4-diacetylphloroglucinol biosynthesis gene phlA_Pf-5 is activated by germinating pea seeds. The 2,4-diacetylphloroglucinol producing Pf-5 derivative, but not its 2,4-diacetylphloroglucinol non-producing mutant, reduced disease severity caused by A. euteiches on pea plants in greenhouse conditions. This is the first report that 2,4-diacetylphloroglucinol produced by strains of Pseudomonas species plays an important role in the biocontrol of pea Aphanomyces root rot. Full article

Amphibians constitute the class of vertebrates with the highest proportion of threatened species, with infectious diseases being considered among the greatest causes for their worldwide decline. Aquatic oomycetes, known as “water molds”, are fungus-like microorganisms that are ubiquitous in freshwater ecosystems and are capable of causing disease in a broad range of amphibian hosts. Various species of Achlya sp., Leptolegnia sp., Aphanomyces sp., and mainly, Saprolegnia sp., are responsible for mass die-offs in the early developmental stages of a wide range of amphibian populations through a disease known as saprolegniosis, aka, molding or a “Saprolegnia-like infection”. In this context, the main objective of the present review was to bring together updated information about saprolegniosis in amphibians to integrate existing knowledge, identify current knowledge gaps, and suggest future directions within the saprolegniosis–amphibian research field. Based on the available literature and data, an integrated and critical interpretation of the results is discussed. Furthermore, the occurrence of saprolegniosis in natural and laboratory contexts and the factors that influence both pathogen incidence and host susceptibility are also addressed. The focus of this work was the species Saprolegnia sp., due to its ecological importance on amphibian population dynamics and due to the fact that this is the most reported genera to be associated with saprolegniosis in amphibians. In addition, integrated emerging therapies, and their potential application to treat saprolegniosis in amphibians, were evaluated, and future actions are suggested. Full article

The pea (Pisum sativum L.) is an excellent protein source for livestock and human nutrition. However, its growth is hampered by several factors including powdery mildew (Erysiphe pisi DC) and lodging in irrigation areas. These limitations may be solved through combining a Canada powdery mildew-resistant pea (Mp1807) with green cotyledon cultivar (Graf) by means of sexual hybridization. A bred semi-leafless pea, cultivar Longwan No. 6, was selected and evaluated in multiple ecological zones to investigate and evaluate high-yield, adaptability, and resistance to root rot (Aphanomyces euteiches Drechs. f. sp. pisi) and powdery mildew from 2012 to 2014. The results revealed that Longwan No. 6 was characterized by lodging resistance and moderate powdery mildew resistance. The average grain yield of Longwan No. 6 is 2855.3 kg/hm² in spring sowing and is increased by 18.4% compared to the control cultivar (Longwan No. 1). The average grain yield of Longwan No. 6 is 2349.9 kg/hm² in winter sowing and is increased by 12.8% when compared to the control cultivar. The results indicated that Longwan No. 6 has a greater potential to increase yield and wide adaptability. Adopting the Longwan No. 6 pea cultivar contributes to improvements in pea production in irrigation areas. Full article