The antibacterial activities of silver (Ag) and copper (Cu) monometallic and bimetallic nanoparticles (MNPs and BNPs) against the most economically important and common plant bacterial diseases in Kyrgyzstan were tested, as well as their phytotoxic effects on wheat and corn seeds. To achieve this, a range of Ag and Cu MNPs and BNPs were synthesized using different methods and were tested at different concentrations to compare their biological effects. The antibacterial activities of Ag and Cu MNPs and Ag‐Cu BNPs against the plant pathogenic bacteria Erwinia amylovora, Pseudomonas syringae, and Pectobacterium carotovorum were evaluated. Of the three pathogens, Ps. syringae was found to be the most sensitive to NPs. The antimicrobial activities of the tested NPs showed significant positive correlations with the concentration; hence, there was a dose‐dependent effect. Due to their antibacterial properties, these Ag‐Cu nanocomposites have high potential in plant protection applications.

(1) Background: The entomopathogenic fungus Metarhizium anisopliae sensu lato forms a species complex, comprising a tight cluster made up of four species, namely M. anisopliae sensu stricto, M. pinghaense, M. robertsii and M. brunneum. Unambiguous species delineation within this “PARB clade” that enables both the taxonomic assignment of new isolates and the identification of potentially new species is highly solicited. (2) Methods: Species-discriminating primer pairs targeting the ribosomal intergenic spacer (rIGS) sequence were designed and a diagnostic PCR protocol established. A partial rIGS sequence, referred to as rIGS-ID800, was introduced as a molecular taxonomic marker for PARB species delineation. (3) Results: PARB species from a validation strain set not implied in primer design were clearly discriminated using the diagnostic PCR protocol developed. Using rIGS-ID800 as a single sequence taxonomic marker gave rise to a higher resolution and statistically better supported delineation of PARB clade species. (4) Conclusions: Reliable species discrimination within the Metarhizium PARB clade is possible through both sequencing-independent diagnostic PCR and sequencing-dependent single marker comparison, both based on the rIGS marker.

In the last two decades, fire blight has progressively spread eastward from Europe and the Mediterranean area to several pome-fruit producing regions of Asia. Its causative agent, the bacterial pathogen Erwinia amylovora, was detected in several new countries, including Georgia, Kyrgyzstan, and Kazakhstan. In the latter two states, the disease creates a threat not only to the commercial apple and pear production, but also to the wild Malus and Pyrus species that constitute the basis of the local forest ecosystems. In this study, we investigated the genetic diversity of the pathogen in Central Asia and the Caucasus region utilizing CRISPR Repeat Regions (CRRs) genotyping and genome sequencing, with the aim to understand its dissemination patterns across the continent. Genome sequence analysis revealed that all strains from these two regions exclusively derived from the archetypal CRR1 genotype A. Our analysis revealed three main E. amylovora clades in Central Asia, with distinct yet partial overlapping geographical distributions. Genomic relationships among isolates indicate that Central Asian strains are genetically closest to those from the Persian region and the Middle East, while the Georgian population is genetically more distant and can align with strains from the Volga District in southern Russia and the Eastern Mediterranean area. Notably, this study also includes strains from the first confirmed occurrences of fire blight in Uzbekistan, Tajikistan, and China. Our findings highlight the importance of phylogenetic analysis and genome sequencing in understanding the phytopathogen epidemics and protecting key agricultural species and the genetic resources of their wild counterparts in the forest.

(1) Background: Potentially arthropod-pathogenic and plant-associated Metarhizium fungi are of high interest for basic research, biological pest control and plant growth promotion. Unambiguous species delineation enabling the taxonomic assignment of new isolates and the identification of new Metarhizium species is of crucial importance for both research and application. Recently, the new species Metarhizium hybridum and Metarhizium parapingshaense were introduced on the basis of phylogenomic studies. (2) Methods: Neighbor- joining and Bayesian inference-based phylogenetic reconstruction of ribosomal intergenic spacer (rIGS) sequences were used to critically evaluate new species introductions. A species-discriminating diagnostic PCR tool for Metarhizium was adapted to M. hybridum and M. parapingshaense. GenBank database mining was performed to identify cryptic descriptions of the new species. (3) Results: The introduction of M. hybridum and M. parapingshaense was corroborated by rIGS sequence comparison. Data mining revealed cryptic first descriptions of M. hybridum from Canada, China, Colombia, Costa Rica, Cuba, Honduras, Mexico, New Zealand, the USA and the Philippines, and of M. parapingshaense from China, India, Japan, the Philippines and South Korea. (4) Conclusions: Results support the reliability of rIGS as a single taxonomic marker for species-level identification of Metarhizium fungi. Species-discriminating diagnostic PCR was successfully adapted to enable the sequencing-independent identification of the confirmed new species M. hybridum and M. parapingshaense.

Soil bacterial and fungal communities were investigated in relation to soil type and farm management practices after vegetation harvesting in autumn. Soils from fields cultivated with Phaseolus vulgaris (bean) and Pyrus comminus (pear) and nonarable, natural areas were studied. Microbial diversity was analysed using cultivation-dependent methods (isolation of pure cultures) and cultivation-independent methods (direct extraction of DNA from soil, followed by PCR amplification of the 16S rRNA and 18S rRNA genes). The use of cultivation-dependent methods revealed that there were no differences in the biodiversity of the soil bacterial and fungal communities between fields cultivated with bean plants and pear trees. However, the use of cultivation-independent methods showed that there were clear soil and crop type-specific effects on the composition of the soil bacterial and fungal communities. The density of the bacterial population was two times higher in northern mountain-valley serozem (NMVS) soil samples than in light chestnut (LC) soil samples. In contrast, the densities of the fungal communities were almost equal in the studied soil types. The density of the actinomycetes community was almost two times higher in LC soil than in NMVS soil under bean plants. The Shannon index values showed that the bacterial biodiversity in the NMVS soil samples was greater than that in the LC soil samples. Soils under fallow appeared to have diverse bacterial communities that mainly consisted of local autochthonous microflora and a small amount of zymogenic microflora (since fresh plant residue does not enter the soil). The Shannon index results revealed two interesting facts: (1) the soil bacterial community was highly diverse in soils that supported bean plants and (2) the soil fungal biodiversity was high under pear trees in both soil types.