Kazakhstan’s rich biodiversity includes diverse apple populations, notably the wild apple tree (Malus sieversii) prized for traits like disease resistance and adaptability, potentially aiding breeding programs. Analyzing their microbiomes offers insights into bacterial diversity and how it influences apple tree development, making it a reliable method for understanding ecological interactions. In this research, 334 apple tree samples were collected from different mountain ranges in southeastern Kazakhstan. An analysis using nanopore-based 16S rRNA sequencing showed a distinct similarity in the microbiome compositions of samples from the Zhongar and Ile Alatau mountain ranges, with a predominance of Pseudomonadaceae, Enterobacteriaceae, and Microbacteriaceae. In contrast, samples from Ketmen ridge showed a higher prevalence of Enterobacteriaceae. Alongside the less represented Pseudomonadaceae family, in the Ketmen ridge region, bacteria of the Xanthomonadaceae, Alcaligenaceae, and Brucellaceae families were also present. Across all regions, beneficial plant-associated bacteria were identified, such as Pseudomonas veronii, Stenotrophomonas geniculata, and Kocuria rhizophila, potentially enhancing plant resilience. However, opportunistic phytopathogens were also detected, including Pseudomonas viridiflava and Serratia marcescens, particularly in the Ile Alatau region. These findings highlight the complex microbial interactions in M. sieversii, thus offering key insights into host—microbe relationships that can inform apple breeding and ecological preservation efforts.

The spongy moth (Lymantria dispar) is a significant defoliator of broadleaf forests and fruit trees, but its population genetic structure in Central Asia remains poorly defined. We analyzed 153 specimens collected from ten populations across Kazakhstan between 2023 and 2024, using a combination of diagnostic real-time PCR, RAPD genotyping, and mitochondrial COI sequencing. Subspecies identification and Asian introgression were initially assessed with the COI/FS1 TaqMan assay. All samples carried the European mitochondrial type L. dispar dispar, while the nuclear FS1 locus indicated Asian introgression in 92.8% of specimens, including eleven heterozygotes for the Asian allele. RAPD amplification with five polymorphic primers produced 107 variable bands, which we converted into a presence/absence matrix for population genetics analysis. Principal coordinate analysis showed weak but detectable population structure, and Bayesian clustering (STRUCTURE) supported the presence of two primary nuclear clusters. Populations from Ketpentau, Sumbe, Kazachka, Butakovka, and Pavlodar were assigned predominantly to one cluster, whereas most individuals from Almaty, Ile-Alatau, Koksu, North Kazakhstan, and Tekeli were assigned predominantly to the other, with limited admixture in several localities. COI sequences formed a single, minimally differentiated L. dispar clade, with no evidence of distinct mitochondrial lineages. Overall, these findings reveal extensive Asian nuclear introgression in populations retaining European mitochondrial haplotypes, supporting the interpretation of Kazakhstan as part of a broad contact zone within the larger dispar–asiatica gradient. The observed mito-nuclear discordance has important implications for quarantine diagnostics and highlights the need for multilocus molecular assays in regional biosurveillance.

The present study provides a comprehensive account of the advancement and utilization of guide RNAs (gRNAs) through the utilization of CRISPR/Cas systems in the identification of two prominent apple viruses, namely Apple Chlorotic Leaf Spot Virus (ACLSV) and Apple Stem Pitting Virus (ASPV). The presence of these viruses presents significant risks to the worldwide apple production industry, due to their adverse effects on fruit quality and yield. The primary objective of this study is to investigate the utilization of CRISPR technology, namely the CRISPR/Cas12a and Cas13a systems, when employed for accurate and highly sensitive diagnostic applications. The present methodology focuses on exploiting conserved areas within the viral genome to improve the efficacy of detection.The methodology involves the development of guide RNAs (gRNAs) that selectively bind to con-served areas within the viral genomes of ACLSV and ASPV. This is achieved by the application of bioinformatics techniques, which aim to reduce unintended effects and enhance the specificity of the targeting. The first stage of in vitro testing entails the production of synthetic viral RNA, followed by live plant testing to verify the efficacy of the gRNAs in real-world settings. Furthermore, the research project includes the generation of complete cDNA clones of the viruses in order to enhance comprehension of their genomic architectures, a critical aspect in the improvement of gRNA designs.This study highlights the incorporation of sophisticated genomic methodologies alongside conven-tional virology approaches in order to enhance the identification and control of plant viral illnesses. The objective is to enhance agricultural well-being and productivity by develop novel diagnostic strategies.

The article proposes and tests an integrated technical scheme for the early detection, digital passportization, and preparation of Halyomorpha halys Stål samples for molecular-genetic verification under the conditions of southern Kazakhstan. The aim of the study was to develop a reproducible system combining field monitoring, GPS referencing of foci, standardized recording of samples, and the formation of a collection suitable for subsequent analysis using the COI and ITS1 markers. Field testing of the system was carried out in the Botanical Garden of Almaty using pheromone traps and trap belts on apple and pear trees. During the observations, 215 individuals at different developmental stages were recorded. The highest abundance was detected in the central part of the surveyed territory; the maximum values were recorded at points D1 and D2 (30 and 33 individuals, respectively). It was established that pheromone traps provide more complete detection of adult imagoes, whereas trap belts increase the detectability of nymphal stages. A unified digital passport of the sample was proposed, ensuring the traceability of data from the moment of collection to the stage of laboratory processing and spatial analysis. The developed system can serve as the first stage of dissertation research on the molecular structure of invasive populations of H. halys and as a basis for regional phytosanitary monitoring programs.

This article examines the prevalence and biological development of the oriental codling moth (Grapholita molesta (Busck)), a quarantine pest that causes significant damage to plants in southeastern Kazakhstan. Field observations and analysis of the phytosanitary condition of plantings revealed that in southeastern Kazakhstan, the pest can develop up to five generations during the growing season, damaging the shoots and fruits of stone and pome fruits. The most vulnerable fruit crops are peach (Prunus persica), plum (Prunus salicina), apricot (Armeniaca vulgaris), and, to a lesser extent, apple (Malus domestica). The highest prevalence of the oriental codling moth was observed in peach and plum plantings, where an average of 25–35 males were caught per trap per week during their mass flight period. It was noted that shoot damage leads to drying out and reduced growth, while fruit damage causes a loss of marketability and premature fruit drop. Shoot damage by the pest reached 18.2 %, and fruit damage reached 22.8 %. It was established that increased air temperature leads to earlier phenology of G. molesta, which may contribute to the significant spread of the pest and lead to increased damage in the future. The obtained results allow for the adjustment of the timing of measures depending on objectively changing annual conditions and the development of an effective pest protection system for trees. Despite its focal distribution, the oriental codling moth requires systematic population monitoring, the use of integrated plant protection methods, and enhanced quarantine measures to prevent its further spread in Kazakhstan. The practical significance of these results lies in their potential use in developing regional fruit crop protection systems.

Halyomorpha halys (brown marmorated stink bug) is an invasive polyphagous pest that causes significant economic damage to fruit, vegetable, and horticultural crops. The species' high reproductive capacity, broad range of host plants and adaptation to diverse climatic conditions have led to its rapid spread across many countries. The entomopathogenic fungus Beauveria bassiana is a promising biological pest control agent due to its high pathogenicity and environmental safety. The aim of this study was to evaluate the efficacy of a native isolate of B. bassiana against various stages of H. halys. Under laboratory conditions, eggs, nymphs, and adults were treated with conidial suspensions at concentrations of 1×10⁵–1×10⁸ conidia/ml. Mortality rates, infection dynamics, LT₅₀ and LT₉₀ values, and the sublethal effects of the fungus were studied. Results showed that mortality also increased with increasing conidia concentrations. The greatest effect was observed at a concentration of 1×10⁸ conidia/ml: nymph mortality reached 98%, adult mortality 95%, and egg mortality 74%. Furthermore, at the highest concentration, the LT₅₀ index was 3.7 days, indicating the high virulence of the local isolate. The fungus also reduced feeding activity, decreased movement intensity, and impaired reproductive performance. These results demonstrate that the local isolate of Beauveria bassiana exhibits high entomopathogenic activity against Halyomorpha halys and holds promise for use in integrated biological pest management programs.