Dispersal mechanisms of phytopathogenic fungi and oomycetes are critical components of forest disease dynamics. While wind and water are well-studied pathways, the role of large forest herbivores as passive vectors remains significantly overlooked. This study quantifies and compares the pathogen loads carried on the hooves and hair of wild red deer (Cervus elaphus) to evaluate their epidemiological potential. Swab samples were collected from the hooves and hair of harvested deer in the Czerwony Bór Forest District, Poland. Quantitative PCR (qPCR) assays targeting the ITS1 region were deployed to detect total fungal DNA, Alternaria alternata, Fusarium avenaceum/F. tricinctum, and several Phytophthora species. A linear mixed-effects model was implemented to statistically evaluate variations in pathogen loads across anatomical sampling locations while controlling for individual animal variability. Fungal DNA was detected in 87.5% of hoof samples, showing significantly lower Ct values (13.85–18.54) compared to fur samples (17.02–29.56), which exhibited a more patchy distribution (p = 0.016). Similarly, A. alternata transfer was highly favored by hooves (p < 0.001). Conversely, F. avenaceum was more frequently detected on hair. Among oomycetes, Phytophthora pseudosyringae was detected in all sampled animals, whereas Phytophthora cactorum occurred rarely, and other tested Phytophthora species were not detected. Wild deer carry DNA of multiple fungal and oomycete pathogens and may act as potential passive carriers within forest ecosystems. Hooves constitute the primary vector for soil-borne pathogens due to sustained contact with topsoil, whereas hair facilitates the movement of specific canopy or airborne taxa. These findings suggest that wildlife movements should be considered in future forest biosecurity assessments for comprehensive forest health management and for understanding pathogen exchange between forest and agricultural ecosystems.
Silver nanoparticles (AgNPs) are increasingly applied in agriculture and related technologies due to their antimicrobial properties, yet their interactions with soil-associated organisms and microbial communities remain insufficiently characterized. This study examined the effects of AgNP exposure (10.85 mg/L) on trace element accumulation and gut bacterial communities of the earthworm Eisenia fetida under two substrate conditions (horticultural substrate and compost). High-throughput 16S rRNA gene sequencing revealed substrate-dependent shifts in microbial community structure following AgNP exposure. Several bacterial taxa, including Proteobacteria, Gammaproteobacteria, Bacilli, Streptococcus sp., and Staphylococcus sp., exhibited pronounced numerical declines, indicating sensitivity to AgNPs, whereas Actinobacteria and Bacteroidetes showed comparatively higher relative abundances, suggesting greater tolerance. Compost partially mitigated the inhibitory effects of AgNPs on gut microbiota. Concurrently, AgNP exposure altered trace element accumulation patterns in earthworm tissues, highlighting interactions between silver uptake and elemental homeostasis. Collectively, these findings demonstrate that AgNPs can induce taxon- and substrate-specific responses in earthworm-associated microbial communities and metal accumulation, providing insight into potential ecological consequences of nanoparticle use in agricultural systems.
Accurate species identification is essential for biodiversity studies, ecological monitoring, and biosecurity, but current molecular approaches rely on PCR amplification, which requires universal primers. Here, we evaluated the possibility of primer-free species identification via direct RNA sequencing (dRNA-seq) by using Oxford Nanopore Technology (ONT). Total RNA isolated from three major eukaryotic kingdoms—fungal (Tuber magnatum Picco 1788), plant (Corylus avellana L. 1753), and animal (Eisenia fetida Savigny 1826) specimens—was sequenced without reverse transcription and PCR amplification. Taxonomic assignments based on similarity to ribosomal nuclear (fungi and animals) and chloroplast (plant) transcripts enabled reliable host species identification. Ribosomal RNA reads dominated the datasets and supported accurate identification of the target organisms, while additional sequences revealed associated microbiota and co-occurring taxa. Notably, dRNA-seq successfully detected symbiotic bacteria in E. fetida and latent fungal infection in C. avellana, highlighting the method sensitivity. However, the complex organization of nuclear ribosomal gene clusters may complicate taxonomic assignment in plants, underscoring the need for improved analytical pipelines. Overall, our results provide proof of concept that dRNA-seq enables primer-independent species identification while simultaneously providing insights into the host microbiomes. Direct sequencing of naturally present RNA molecules generated sufficient sequence information for species identification across different taxonomic kingdoms, indicating the technical feasibility of the developed technique. Further validation using larger biological datasets and direct comparisons with conventional sequencing methods will determine its wider applicability as an innovative and complementary species identification approach.
Substituting peat moss with compost derived from organic waste in plant nurseries presents a promising solution for reducing environmental impact, improving waste management, and enhancing soil health while promoting sustainable agricultural practices. However, selecting the appropriate proportions of both materials is crucial for each plant species. This study investigates the effects of different ratios of compost and peat mixtures on the growth and development of pepper seedlings. The compost mixtures used in the study included the following combinations: sewage sludge with sawdust (A), sewage sludge with sawdust and biodegradable garden/park waste (B), and biodegradable garden/park waste with sawdust (C). The final substrates used for seedling production were composed of composts (A, B, C) and peat (O) as a structural additive, mixed in different proportions by mass: I-O 25%, II-O 50%, and III-O 75%. Seedlings grown in these substrates were assessed using biometric and physiological measurements. Nematode species present in substrates were identified by metabarcoding analysis. The results revealed that substrate productivity depended not only on nutrient content but also on structural properties, which were significantly influenced by the peat proportion. Among the tested compost mixtures, variant A I emerged as the most effective substrate, promoting optimal seedling growth. Molecular nematode analysis revealed significant nematode contamination in substrates with higher peat proportions (C II and C III), including Meloidogyne sp. Lichtenburg (26%), Meloidogyne hispanica (5%), Meloidogyne sp. Mi_c1 (3%), Meloidogyne ethiopica (2%), and Meloidogyne thailandica (1%). The findings underscore the critical importance of achieving an optimal balance between nutrient content and structural properties in substrates to support the healthy growth and development of pepper seedlings. To further enhance crop performance and reduce the risk of pest-related damage, it is essential to prioritize the improvement of substrate selection strategies. Monitoring for nematode contamination is crucial to prevent potential compromises in seedling quality and overall productivity.
Forensic entomology is concerned with the study of arthropods that assist in criminal investigations. Among them, necrophagous beetles (Coleoptera) play a crucial role as they are found in all stages of decomposition of corpses, which makes them valuable for forensic analysis. Accurate species identification is crucial for the effective use of entomological evidence. Efficient, cost-effective, and rapid methods are essential for this purpose. In the present study, we developed a novel assay that enables simple, inexpensive, and rapid identification of necrophagous beetle species. Using two primer sets targeting the cytochrome c oxidase 1 (COI) gene and analyzing the resulting SYBR Green I melting curves, we were able to identify fourteen beetle species: Aleochara curtula (Goeze, 1777), Anoplotrupes stercorosus (Scriba, 1791), Creophilus maxillosus (Linnaeus, 1758), Dermestes undulatus (Brahm, 1790), Hister unicolor (Linnaeus, 1758), Margarinotus brunneus (Fabricius, 1775), Necrodes littoralis (Linnaeus, 1758), Nicrophorus investigator (Zetterstedt, 1824), Ontholestes murinus (Linnaeus, 1758), Oiceoptoma thoracicum (Linnaeus, 1758), Philonthus cognatus (Stephens, 1832), Saprinus planiusculus (Motschulsky, 1849), Silpha tristis (Illiger, 1798), and Thanatophilus rugosus (Linnaeus, 1758). Data on the melting temperature of the amplicon can be obtained within 60-90 min after DNA extraction. This study expands the understanding of the species composition of necrophagous Coleoptera, which is crucial for forensic entomology methods based on insect succession. Furthermore, it provides the first comprehensive data on necrophagous beetles in the vicinity of Kampinos National Park.
Earthworms are important in agriculture in the process of soil fertilization and influence its physicochemical properties. The taxonomic classification of earthworms using morphological characteristics requires experts, is difficult, and can require specimen dissection to extract detailed anatomical studies. Molecular techniques are time-consuming and expensive. The objective of this study was to distinguish earthworms belonging to different genera, Eisenia, Dendrobaena, and Lumbricus, using an innovative approach involving machine learning models built based on image texture parameters from individual color channels R, G, B, L, a, b, X, Y, Z, U, V, and S. The earthworms Eisenia fetida, Dendrobaena ssp., and Lumbricus terrestris were used as research materials. Image acquisition was performed using a flatbed scanner on a black background. In the case of each earthworm, 2172 texture parameters from images in individual color channels R, G, B, L, a, b, X, Y, Z, U, V, and S were extracted. Textures after selection were used to develop classification models using machine learning algorithms. The earthworms Eisenia fetida, Dendrobaena ssp., and Lumbricus terrestris were distinguished with the accuracy reaching 100% for models built using Logistic, Ensemble, and Narrow Neural Network. All earthworms were correctly classified. Also, in the case of other models, earthworm classes were distinguished with high accuracies, such as 99% (Naive Bayes, Random Forest, SVM, KNN), 97% (Simple Logistic), and 94% (KStar). For the most important species, E. fetida, the correctness of the species identification was confirmed by direct RNA sequencing. The application of image analysis and machine learning turned out to be a non-destructive, inexpensive, and objective approach to distinguishing earthworms belonging to different genera.
True slime molds (Eumycetozoa) represent a monophyletic clade within the phylum Amoebozoa, comprising the lineages Myxogastria, Dictyostelia, and Protosporangiida. Although historically misclassified as fungi, recent molecular and biochemical studies underscore their distinct evolutionary trajectories and rich metabolomic profiles. In this review, we synthesize current knowledge on Eumycetozoa as a reservoir of bioactive compounds, detailing how secondary metabolites—including polysaccharides, amino acids, unsaturated fatty acids, terpenoids, and glycosides—vary across plasmodia, fruiting bodies, and spores. A systematic literature search in major scientific databases accounted for legacy nomenclature and leveraged chemoinformatic tools for compound verification. Our findings reveal 298 distinct metabolites that serve ecological roles in nutrient recycling and interspecies interactions, while also showing promise for controlling agricultural pests and pathogens. Notably, certain glycosides, lectins, and polyketides exhibit antimicrobial or cytotoxic activities, indicating their potential utility in managing these biological challenges. By consolidating current data and emphasizing the wide taxonomic range of Eumycetozoa, this review highlights the critical need for comprehensive biochemical and genomic investigations. Such efforts will not only advance our understanding of slime mold metabolomes and their evolutionary significance but also pave the way for innovative, eco-friendly applications.
The casein gene cluster spans 250 to 350 kb across mammalian species and is flanked by non-coding DNA with largely unknown functions. These regions likely harbor elements regulating the expression of the 4 casein genes. In Bovidae, this cluster is well studied in domestic cattle and to a lesser extent in zebu and water buffalo. This study used a cattle-specific SNP microarray to analyze 12 Bovidae taxa and estimate casein gene cluster variability across 5 bovid subfamilies. Genotyping identified 126 SNPs covering the entire casein gene cluster and 2 Mb of upstream and downstream regions. Dairy cattle, watusi, and zebu showed the highest polymorphism: 63.7–68.2% in the 5′-upstream region, 35.6–40.0% in the casein cluster, and 40.4–89.4% in the 3′-downstream region. Among wild bovids, only a ‘semi-aquatic’ lechwe revealed high polymorphism similar to cattle. Other species exhibited lower variability, ranging from 9.1–27.3% in the 5′-upstream, 8.9–20.0% in the casein, and 4.2–10.6% in the 3′-downstream regions. For the first time, genome variability data were obtained for impala, waterbuck, and lechwe. It appears that higher variability in cattle’s casein gene cluster may relate to its intense expression. This study confirms the effectiveness of cattle-derived microarrays for genotyping Bovidae.
Vermicomposting stands as a sustainable and environmentally friendly waste management practice, leveraging the metabolic prowess of earthworms to facilitate the decomposition of organic matter into nutrient-rich compost. The aim of this experiment was to study the influence of Eisenia fetida on the density and the tropic structure of a nematode community during the vermicomposting process over a period of 3 months. Sewage sludge and green waste served as composting substrates. Overall, six compost variants were prepared consisting of three variants incorporating E. fetida and three control variants lacking E. fetida. Throughout the investigation, samples were gathered on a monthly basis, with each variant undergoing three repetitions. The aim was to isolate nematodes, determine the population density of the five trophic groups, and identify the dominant community. The anaysis was conducted employing both microscopic examination and molecular metabarcoding (NGS). It was shown that the bacterial-feeding community maintained dominance. The introduction of E. fetida into the compost led to a significant rise in the abundance of Diplogasteridae. In the variant without E. fetida, the amount of Diplodasteridae exceeded 0.1% only after the 30th (C3) or the 60th (C1, C2) day of composting, while in the compost with E. fetida, they were present in large amounts (ranging from 11.0% to 28.0%) already on the 30th day of composting. The introduction of Eisenia fetida also led to a notable reduction in H. gingivalis with significant implications for mitigating the risk of halicephalobiosis. The introduction of E. fetida resulted in reducing H. gingivalis to levels below 0.1% in all compost variants.
The spread of mistletoe, which mainly affects Scots pine in Europe’s managed forests, has become a serious ecological and economic problem, exacerbated by climate change and drought stress. Research into mistletoe-associated mycobiota provides crucial insights into the biology, ecology and distribution of these fungi and forms the basis for sustainable plant protection and effective management strategies. The aim of this study was to identify the fungal species associated with mistletoe (Viscum album L.) at 75 sites across Poland, with a focus on the northeastern regions of the country. The field work included a systematic and targeted search using the transect method, followed by laboratory analysis, during which fungal samples were prepared and examined under both light and scanning electron microscopes. The genetic analysis was carried out using ITS sequencing to confirm the presence of pathogen-specific fungi in the majority of V. album samples examined. The results showed four fungal species associated with the mycobiota of mistletoe: Sphaeropsis visci, Plenodomus visci, Cylindrodendrum hubeiense and Septoria krystynae. It is noteworthy that two of these species (P. visci and C. hubeiense) have not yet been detected in Poland. Based on these findings, the distribution areas of the identified fungal species were mapped accordingly.
Background: Climate change is leading to severe and long-term droughts in European forest ecosystems. can have profound effects on various physiological processes, including photosynthesis, gene expression patterns, and nutrient uptake at the developmental stage of young trees. Objectives: Our study aimed to test the hypothesis that the application of silica (SiO2) influences photosynthetic efficiency and gene expression in 1- to 2-year-old Fagus sylvatica (L.) seedlings. Additionally, we aimed to assess whether silicon application positively influences the structural properties of leaves and roots. To determine whether the plant physiological responses are genotype-specific, seedlings of four geographically different provenances were subjected to a one-year evaluation under greenhouse conditions. Methods: We used the Kruskal-Wallis test followed by Wilcoxon's test to evaluate the differences in silicon content and ANOVA followed by Tukey's test to evaluate the physiological responses of seedlings depending on treatment and provenance. Results: Our results showed a significantly higher Si content in the roots compared with the leaves, regardless of provenance and treatment. The most significant differences in photosynthetic performance were found in trees exposed to Si treatment, but the physiological responses were generally nuanced and provenance-dependent. Expression of hsp70 and hsp90 was also increased in leaf tissues of all provenances. These results provide practical insights that Si can improve the overall health and resilience of beech seedlings in nursery and forest ecosystems, with possible differences in the beneficial role of silicon application arising from the large differences in wild populations of forest tree species.
In this study, greenhouse tests were conducted on 240 Fraxinus excelsior seedlings to investigate the simultaneous damage caused by thea pathogenic fungus and oomycetes. The experiment was performed under controlled conditions in the greenhouse of the Institute of Forest Research in Sękocin Stary (Poland). Three species of oomycetes were used for the experiment: Phytophthora plurivora, Phytophthora taxon hungarica, Phytophthora megasperma, and the fungus Hymenoscyphus fraxineus. Inoculations using the fungus were carried out on shoots and in plant pots in which the soil was mixed with the three Phytophthora species mentioned above, both simultaneously and separately, which made it possible to recognize the cumulative effect of the related plant infection. The aim of the study was to investigate the effect of phosphite-containing preparations on the health of common ash under conditions of threat to the roots by Phytophthora spp. and damage to the aerial parts of the plant by the fungus, as well as the possible occurrence of synergistic effects. Two types of protective preparations (Actifos and Phos60 of the nitrogen and potassium forms, respectively) were used. It was found that the inoculation of ash seedlings with the fungus H. fraxineus resulted in plant mortality, while the mixture of Phytophthora did not cause significant damage. It was confirmed that when pathogens coexist, a phenomenon occurs that leads to an acceleration in the development of disease symptoms and, thus, to plant mortality. In vitro tests confirmed the usefulness of phosphite preparations for the protection of ash seedlings.
Meloidogyne hapla is one of the most important nematode pathogens. It is a sedentary, biotrophic parasite of plants that overwinters in the soil or in diseased roots. The development of M. hapla is temperature dependent. Numerous studies have been performed on the effect of temperature on the development of M. hapla, but only a few of them analyzed the heat shock protein (hsp) genes. The aim of the study was to perform expression profiling of eight hsp genes (Mh-hsp90, Mh-hsp1, Mh-hsp4, Mh-hsp6, Mh-hsp60, Mh-dnj19, Mh-hsp43, and Mh-hsp12.2) at two development stages of M. hapla, i.e., in eggs and second-stage juveniles (J2). The eggs and J2 were incubated under cold stress (5 °C), heat stress (35 °C, 40 °C), and non-stress (10 °C, 20 °C, and 30 °C) conditions. Expression profiling was performed by qPCR. It was demonstrated that only two genes, Mh-hsp60 and Mh-dnj19, have been upregulated by heat and cold stress at both development stages. Heat stress upregulated the expression of more hsp genes than cold stress did. The level of upregulation of most hsp genes was more marked in J2 than in eggs. The obtained results suggest that the Mh-hsp90 and Mh-hsp1 genes can be used as bioindicators of environmental impacts on nematodes of the Meloidogyne genus.
In the last two decades bovine beta-casein variant A1 has attracted attention in the dairy industry as a source of beta-casomorphin-7 opioid-like peptide with potentially harmful effects on human health. Because the origin of this variant is unknown, the aim of the study was to investigate its occurrence in different species of the Bovidae family. Blood samples were taken from 137 animals belonging to 17 Bovidae species. All animals were genotyped in beta-casein locus by Illumina Bovine MDv2 Chip. The results shown that allele A1 was present in Bos taurus cattle, including primitive cattle (Watusi and Ukraine) and in a modern dairy breed (Holstein). In Dahomey cattle, two heterozygous A1A2 individuals were found. In all remaining species, no animal carrying allele A1 of beta-casein was identified. It may be concluded that allele A2 is ancestral to allele A1 and was spread especially within modern dairy cattle
Biodiversity within composting systems involves a variety of microorganisms including nematodes. In the research, nematode populations were monitored within six simultaneously operating composting processes. These processes involved varying proportions of feedstock materials. The primary objective was to evaluate the consistency of nematode community succession patterns across the composting processes over a time of 3 months. During the study, samples were taken every month to isolate nematodes, determine the population density of the five trophic groups (per genus) and determine the dominant nematode species. It was shown that the bacterial-feeding community maintained dominance, while the fungus-feeding nematodes gradually increased in dominance as the maturation process progressed. The presence of predatory nematodes Mononchoides which were initially absent, along with the total absence of parasitic nematodes in the late stages of waste stabilization, serves as strong evidence for the reliable evaluation of the biodegradable waste processing level. Based on the obtained results, it is evident that the succession of nematode communities holds promise as a reliable method for evaluating compost maturity.
Agriculture can degrade soils and reduce microbial diversity. The reduction in microbial diversity of degraded soils is due to their long-term agricultural use. In most cases, such areas are afforested but rarely succeed in converting them into first-generation pine forests without adequately revitalizing the soils and restoring the natural relationships characteristic of forest habitats. This is possible thanks to the positive changes in soil biodiversity. To facilitate and enhance this phenomenon in the present experiment, various forms of organic matter were added to the soil: pine bark compost scattered on the soil surface (BCS) or placed under the roots of the seedlings at planting (BCR), woody debris (WW) and sawdust (S), while the control plot was not treated. The studies started in 2001 on two experimental plots, a poorer one in Bielsk and a more fertile one in Czarne Człuchowskie (eastern and northern Poland, respectively). The following year, 2-year-old pine seedlings (Pinus sylvestris L.) were planted on the plots. After 20 years, the physicochemical properties of the soil and its microbial composition were determined and compared with the control. The results encourage the use of organic matter for established pine forest crops on post-agricultural land for revitalization: C, N, and P content increased both in the organic layer and in the topsoil (up to 40 cm), where most fine roots are located. The total content of exchangeable base cations (Ca, Mg, K) and the sorption capacity of the soils (a measure of the ion binding capacity of the soil) also improved. The genetic analyses carried out using the molecular method (NGS) showed positive changes in the composition of the soil microbiome. Compared to poorer soil conditions in richer habitats, the number of taxa increases when organic matter is added, leading to significant qualitative changes in the bacteriome. The addition of organic material from the forest had a positive effect on the bacterial communities, which in turn accelerated the changes in the diversity of bacteriomes characteristic of agricultural soils and brought them closer to the forest ecosystem. The organic horizon was restored, and at the same time, the biodiversity of the soil microbiome increased, which is important for the health and sustainability of pine stands on formerly agricultural land.
A 20-year study of a pine stand on post-agricultural land showed that woody debris in the form of organic matter can be successfully used to restore symbiotic mycorrhizal communities, as is the case with forest soils. Woody substrates restored organic matter in soils altered by long agricultural use and had a positive effect on the composition of mycobiota antagonistic to pathogens, especially to Heterobasidion annosum, the causal agent of the dangerous disease root and stump rot of many forest tree species, including stands of Pinus sylvestris (L.). In a study that started in 2001 in the forest district of Czarne Człuchowskie (northern Poland), the following organic materials were used: wood residues (W), sawdust (S), bark compost (B), and compost applied to the root zone during planting (G). The organic materials were spread in the form of mulch over the entire area during planting. After twenty years, it was found that the substrates used provided suitable growth conditions for mycobiome useful for pines. The addition of organic matter did not change the alpha biodiversity of the soil, but in the long term led to significant changes in the composition of mycobiota (beta biodiversity). The changes in the soil after the addition of organic material naturally accelerated the formation of the forest habitat. A number of fungi evolved that degraded added lignin and cellulose while being antagonists of H. annosum and other pine pathogens. In particular, the well-known hyperpathogens of the genus Trichoderma played an important role by promoting resistance of the soil environment to pathogens. Soil enrichment by bark compost and wood residues increased the relative abundance of Trichoderma more than fourfold.Mycorrhizal fungi became dominant in soil enriched with organic matter. After enriching the soil with bark compost, the relative abundance of Amphinema and Inocybe increased to 5%. The relative abundance of Russula in soil enriched with wood residues and sawdust increased to 9% and 5%, respectively. Mycorrhizal fungi, e.g., of the genus Ąmanita, Rusula, which formed root mycorrhizae, not only increased the root receiving area many times over, but also protected the roots (mechanically and chemically from pathogens). Altogether, the observed positive changes increase the chances that the first generation of pines will survive on the ground.
Betula pendula Roth. is considered a pioneering plant species important for urban ecosystems. Based on the sequencing of fungal ITS, we characterized the ectomycorrhizal (ECM) communities of twenty silver birch trees growing in a contaminated, highly anthropo-pressured urban environment and in a natural reserve site. We analysed chemical properties of each tree soil samples, focusing on effects of anthropogenic transformation. Three effects of urbanization: high heavy metal content, increased salinity and soil alkalinity, were highly correlated. The examined trees were divided into two forest and two urban clusters according to the level of anthropogenic soil change. The effect of soil transformation on the ECM communities was studied, with the assumption that stronger urban transformation leads to lower ECM vitality and diversity. The results of the study did not confirm the above hypothesis. The ECM colonization was above 80% in all clusters, but the forest clusters had significantly higher share of vital non-ECM root tips than the urban ones. Eleven mycorrhizal fungal species were identified varying from seven to nine and with seven species observed in the most contaminated urban plot. However, the lowest Shannon species diversity index was found in the most natural forest cluster. In conclusion, our findings demonstrate no significant negative effect of the urban stresses on the ECM communities of silver birch suggesting that both forest and urban trees have the potential to generate a similar set of ECM taxa.
Assuming that the seeds of Vicia sativa L. have a stressful effect on J2 stage Meloidogyne hapla, we undertook research on the effect of these seeds on the motility and mortality of J2 and determined the expression levels of selected hsp genes in J2. The assessment of the effect of V. sativa seeds on the motility of M. hapla specimens consisted of observing the movement of J2 immersed in a seed diffusate or in a tomato root filtrate at temperatures of 10, 17, and 21°C. In J2 treated with V. sativa (cv. Ina) seed diffusates, the expression level of hsp genes was determined by qPCR. J2 exposed to V. sativa diffusates were found to lose their motility, while their mortality did not exceed 30%. J2 in the seed diffusate were characterized by an increase in the expression levels of the Mh-hsp90, Mh-hsp1, and Mh-hsp43 genes. It is suggested that the hsp90 gene may be a potential bioindicator of the environmental impact on Meloidogyne nematodes. The impaired ability to move in J2 of M. hapla is attributable to the occurrence of V. sativa seeds in their habitat. These studies may contribute to developing methods of reducing crop damage caused by M. hapla.
The genus Fusarium comprises more than 300 species, and many of them are pathogens that cause severe diseases in agricultural, horticultural and forestry plants in both antropogenic and natural ecosystems. Because of their importance as plant pathogens, the genomes of several Fusarium spp. have been sequenced. Within this genus, Fusarium circinatum is one of the most harmful pathogens of pine trees attacking up to 60 Pinus species. Till now, the genomes of 13 strains of F. circinatum have been sequenced. The strain GL1327 we studied lacks a twelfth chromosome, which allows the study of virulence genes on this chromosome. Although the genome of several strains of F. circinatum has been sequenced, it is still almost completely unannotated, which severely limits the possibilities to further investigate the molecular mechanisms of virulence of Fusarium. Therefore, this study aimed to annotate the 12th chromosome of F. circinatum and integrate currently available resources. In silico annotation of the 12th chromosome of F. circinatum revealed the presence of 118 open reading frames (ORFs) encoding 141 proteins which were predicted using an ab initio gene prediction tool. The InterProScan and SMART analyses identified known domains in 30 proteins and eggNOG additionally in 12 of them. Among them, four groups can be distinguished: genes possibly related to heterokaryon incompatibility (4 genes), regulation of transcription (5 genes), plant cell wall degrading enzymes (7 genes) and trichothecene synthesis (3 genes). This study also integrated data of F.circinatum reference strain CMWF1803 assembled to chromosome level but not annotated with currently best annotated but assembled only to scaffold level strain NRRL 25331.