MLO proteins are a family of transmembrane proteins in land plants that play an important role in plant immunity and host–pathogen interactions, as well as a wide range of development processes. Understanding the evolutionary history of MLO proteins is important for understanding plant physiology and health. In the present work, we conducted a phylogenetic analysis on a large set of MLO protein sequences from publicly available databases, specifically emphasising MLOs from the tomato plant and related species. As a result, 4886 protein sequences were identified and used to construct a phylogenetic tree. In comparison to previous findings, we identified nine phylogenetic clades, revealed the internal structure of clades I and II as additional clades and showed the presence of monocotyledon species in all MLO clades. We identified a set of 19 protein motifs that allowed for the identification of particular clades. Sixteen SlMLO proteins from tomato were located in the phylogenetic tree and identified in relation to homologous sequences from other Solanaceae species. The obtained results could be useful for further work on the use of MLO proteins in the study of mildew resistance in Solanaceae and other plant families.

Cross-ecosystem transmission of plant pathogens from crops to natural forests is increasingly recognized as a key factor in disease emergence and biodiversity loss. Agricultural systems serve as major sources of inoculum, with landscape interfaces—such as crop–forest edges, riparian zones, abandoned orchards, and nursery–wildland transitions—acting as active epidemiological gateways. Biological vectors, abiotic dispersal, and human activities collectively enable pathogen movement across these boundaries. Host-range expansion, recombination, and hybridization allow pathogens to infect both cultivated and wild hosts, leading to generalist and recombinant lineages that survive across diverse habitats. In natural ecosystems, such introductions can alter community composition, decrease resilience, and intensify the impacts of climate-driven stress. Advances in molecular diagnostics, genomic surveillance, environmental DNA, and remote sensing–GIS (Geographic Information System) approaches now enable high-resolution detection of pathogen flow across landscapes. Incorporating these tools into interface-focused monitoring frameworks offers a pathway to earlier detection, better risk assessment, and more effective mitigation. A One Health, landscape-based approach that treats agro–wild interfaces as key control points is essential for reducing spillover risk and safeguarding both agricultural productivity and the health of natural forest ecosystems.

Monilinia fructigena, a causal agent of brown rot in apple and other fruit crops, poses a significant threat to fruit production and postharvest quality in temperate regions. This study reports on the molecular and morphological identification of M. fructigena isolates obtained from symptomatic apple fruits in the Almaty region of Kazakhstan. Nine isolates were characterized through a combination of morphological assessment, real-time PCR, target locus (ITS and TEF1-α gene) sequencing, and whole genome sequencing using nanopore sequencings. Morphological analysis revealed typical features of M. fructigena, including blastoconidia and microconidia. Pathogenicity tests on ‘Idared’ and ‘Golden Delicious’ apples confirmed the high aggressiveness of the isolates, with lesion development observed within 24–48 h post-inoculation. Molecular identification via real-time PCR and target sequencing confirmed all isolates as M. fructigena with high mapping quality and sequence identity. The whole genome sequencing of a representative isolate further validated the species identity based on comparative alignment with Monilinia reference genomes. Thus, the combination of the used traditional and molecular methods allowed us to unambiguously identify the isolated fungus as M. fructigena. This integrative approach enhances the understanding of Monilinia species in Central Asia and supports the implementation of modern molecular tools for phytopathogen surveillance and agricultural biosecurity.

Fire blight, caused by Erwinia amylovora, is one of the most damaging bacterial diseases affecting apple production and the safety of wild Malus sieversii populations in Central Asia. Effective monitoring relies on accurate molecular diagnostics; however, comparative data on commonly used detection methods remain limited for the region. In this study, we evaluated the performance of three molecular assays—LAMP, real-time PCR, and targeted nanopore sequencing of a 16S rRNA gene fragment—using 124 plant samples exhibiting fire blight symptoms collected from 30 sites across Southern Kazakhstan and Kyrgyzstan. The results of LAMP, real-time PCR, and the amplification of 16S sequences were highly consistent with each other. Targeted 16S nanopore sequencing reliably identified E. amylovora in all PCR-positive samples, yielding high read counts and consistent species-level classification, although the analyzed 16S region provided limited resolution for intraspecies variation. Across sampling locations, abandoned orchards represented major reservoirs of infection compared to maintained orchards and wild populations. Our results confirm that all three approaches are robust tools for detecting E. amylovora. These findings support the importance of different molecular diagnostic methods to assist fire blight surveillance in the region.

Apple scab, caused by the fungus Venturia inaequalis, is one of the most widespread and economically significant diseases of apple orchards, leading to reduced photosynthesis, fruit damage, and yield losses of up to 70%. In this study, a survey of 30 wild and cultivated apple populations in Kazakhstan and Kyrgyzstan was conducted, encompassing 302 samples. The pathogen was detected in 8 populations (48 samples), corresponding to an infection rate of 16%. Molecular identification using the EF-1α marker and ITS region sequencing definitively confirmed the presence of V. inaequalis in all positive samples. Phylogenetic analysis showed clear population structuring: isolates from Kyrgyzstan formed a distinct clade close to international lineages, while Kazakh isolates showed high genetic variation. These findings highlight the ongoing presence of V. inaequalis in Central Asia and emphasize the importance of combining morphological, molecular, and phylogenetic methods for effective pathogen monitoring and control.

Fungal pathogens of the genus Monilinia pose a significant threat to fruit production, particularly in temperate regions such as southeastern Kazakhstan. This study investigates the morphological, molecular, and pathogenic characteristics of mixed infections caused by Monilinia fructigena and Monilinia polystroma, two closely related species responsible for fruit rot in apple orchards. Samples from symptomatic apple fruits were collected in the Almaty region, and pure fungal cultures were isolated and characterized. Morphological examination of the isolates revealed typical Monilinia colony characteristics, including dense mycelial growth and sporulation patterns. Experimental inoculation of Pyrus communis cv. ‘Conference’ confirmed high pathogenic potential and aggressive lesion development within 24 hours post-inoculation. Molecular identification using species-specific primers targeting the LAC2 gene confirmed mixed infections in 8 out of 9 samples. This is the first documented co-occurrence of M. fructigena and M. polystroma in Kazakhstan. The findings emphasize the necessity of integrating morphological assessment with molecular diagnostics for accurate pathogen identification. The development and use of specific molecular markers enable early detection, species differentiation, and improved phytosanitary strategies to manage moniliosis in fruit crops.