An important direction in industrial microbiology is the development of probiotic strains with valuable consumer properties. The probiotic industry is currently one of the most rapidly developing segments of the food and pharmaceutical sectors. Stearic (octadecanoic) acid C18:0 is one of the major metabolites present in the cell-free supernatant of the bacterium Streptococcus thermophilus, which is widely used in the production of fermented dairy products, including yogurt and cheese. S. thermophilus affects not only the texture and sensory properties of products, but also exhibits various probiotic effects, including antioxidant activity, modulation of the gut microbiota, inhibition of certain pathogens, and others. It is assumed that a number of probiotic effects exerted by S. thermophilus may be mediated through octadecanoic acid as one of its main metabolites. Octadecanoic acid C18:0, like other long-chain fatty acids, enters the human body via several mechanisms, including protein-mediated transport and passive diffusion across cell membranes. Inside the cell, octadecanoic acid serves not only as a substrate for the synthesis of triglycerides and other complex lipids, but, as shown in cell-based and in vivo models, also acts as a modulator of signaling and stress responses, including those associated with apoptosis. This is an important aspect of the influence of stearic acid on organism functioning, underpinning its anti-inflammatory and potentially anti-tumor effects. However, the molecular genetic mechanisms by which octadecanoic acid acts as a probiotic on the human organism remain insufficiently understood. In the present study, using our previously developed information – software system ANDSystem (employing machine learning and artificial intelligence for automatic extraction of knowledge from scientific texts and databases), we reconstructed gene networks regulating the intrinsic (mitochondrial) and extrinsic (death receptor-mediated) apoptotic pathways in human cells under the influence of stearic (octadecanoic) acid. To search for metabolites produced by probiotic microorganisms that may have beneficial therapeutic properties, we propose an approach that combines gene network reconstruction with differential gene expression analysis. Using this approach, we show that octadecanoic acid produced by S. thermophilus can control the intrinsic and extrinsic apoptotic pathways primarily via regulation of PTGS2 expression; the results indicate that cyclooxygenase-2 is a key regulator mediating the effect of octadecanoic acid on apoptosis-related genes.

The rapid development of high-throughput sequencing technologies has led to an explosive accumulation of high-quality bacterial genome sequence data – their number is approaching three million, and this growth continues. This, in turn, provides additional impetus for the development of technologies for more efficient annotation using analytical methods designed to utilize such large-scale genomic data, as well as for achieving new levels of annotation quality. One such analytical approach is phylogenetic footprinting, which aims to identify motifs corresponding to transcription factor binding sites in the promoter regions of bacterial genomes by comparing corresponding sets of regulatory sequences of orthologous genes in related organisms. The continued accumulation of genomic data has served as the basis for further development of this approach. It has been found that an excessive number of sequences in a set analyzed using phylogenetic footprinting only reduces the accuracy of the method, whereas the inclusion of a sequence selection step in the analyzed set based on data on mutual evolutionary distances improves the method’s performance. In this paper, we propose and implement a further step in the development of the phylogenetic footprinting method. This step involves multiple runs of the selection step described above to generate distinct subsamples, subsequent pipeline runs for each subsample, and statistical analysis of the results obtained from multiple pipeline runs. The proposed approach, implemented in the MotifsOnFly method, improves the robustness of motif recognition results obtained from multiple pipeline runs. The effectiveness of the MotifsOnFly method is demonstrated using the analysis of the well-annotated promoter of the Escherichia coli OmpW gene.

This study addresses the challenge of automated high-throughput phenotyping of wheat spike characteristics using modern computer vision and deep learning methods. Accurate estimation of spikelet number is a key indicator of plant productivity, yet traditional manual counting approaches are labor-intensive, slow, and difficult to scale to large breeding datasets. To overcome these limitations, we propose a spikelet detection strategy based on simplified point annotations, where an expert marks only the centers of spikelets rather than drawing detailed segmentation masks or bounding boxes. This significantly reduces annotation time and lowers the overall cost of preparing training datasets for machine learning models. To determine the most effective way of utilizing such simplified annotations, three computational methods were explored: segmentation of binary masks using a U-Net architecture, density regression based on two-dimensional Gaussian distributions optimized via Kullback–Leibler divergence, and detection of fixed-size bounding regions using the YOLOv8 object detection framework. The models were evaluated on dedicated test datasets using both quantitative metrics (MAE, MAPE) and spatial localization metrics (Precision, Recall, F1 score). The results demonstrate that U-Net-based approaches provide consistently high accuracy in spikelet localization and counting while maintaining robustness to annotation imperfections. In contrast, the YOLOv8-based method showed reduced performance, likely due to the geometric mismatch between fixed-size boxes and the natural elongated shape of spikelets. Overall, the proposed methodology highlights the effectiveness of combining minimalistic point-level annotation with advanced segmentation models for automating phenotyping workflows. This approach has the potential to accelerate breeding programs, enhance the efficiency of largescale phenotypic data collection, and support further development of robust computer-vision tools for plant science applications.

Intraspecific infertility, the nature of which is not always understood, occurs in many eukaryotes. Intraspecific PM hybrid dysgenesis (PM HD) in Drosophila melanogaster manifests in one cross direction as offspring infertility and other genetic disorders due to incompatibility between the maternal cytoplasm and the paternal genome. PM HD is believed to result from a massive transposition of the P-element when the maternal cytoplasm lacks a repressor to block it. In this work, we have investigated the distribution of the P transposon and blood retrotransposon in the reference PM HD strains (Canton-S and Harwich), which have been maintained in different laboratories for several decades. P-element distribution patterns vary among Harwich sub-strains, indicating that the P-element was translocated in these genomes. The rate of movement of the P-element, which was not induced by crosses, is comparable to the rate of movement of other DNA transposons. The distribution pattern of the low-active blood retrotransposon in Harwich sub-strains is more stable than that of the P-element, indicating genetic relatedness between sub-strains. Derivatives of the P-element detected in some Canton-S sub-strains possibly indicate genetic contamination. The significant difference in the blood transposable element distribution pattern in Canton-S sub-strains also indicates genetic heterogeneity among them. Despite the complex genealogy of the studied sub-strains, including cases of possible genetic contamination, and differences in P-element distribution, the ability to express PM HD symptoms is preserved in the studied sub-strains.

Genome-wide association studies (GWAS) have become a standard approach for identifying quantitative trait loci associated with diverse phenotypic traits. Further investigation of the locus – specifically, the search for the causal gene and mutation – may present various challenges. One of the challenges is genetic heterogeneity (or locus heterogeneity), when alleles from different closely located genes can influence the same trait. Recently, using GWAS, we found the qDTF-7 locus on soybean chromosome 3, which is associated with flowering time under Novosibirsk conditions. Initially, we identified GmTOE1, an ortholog of TOE1 (TARGET OF EAT1), a known flowering-time regulator in Arabidopsis, as the most likely candidate gene for this locus. Four major haplotypes were identified in GmTOE1, which are associated with soybean flowering and maturity and are likely to provide soybean adaptation to northern latitudes. However, this gene showed only a very weak association with soybean flowering in the Novosibirsk region compared to the Oryol region, suggesting the presence of another gene within the locus that influences flowering time. We therefore re-analyzed genes in the qDTF-7 locus and identified GmRVE8c, an Arabidopsis RVE8 (REVEILLE 8) ortholog, located ~21 kb upstream of GmTOE1; RVE8 is a circadian clock component involved in plant development. After studying the natural variation of the GmRVE8c genes, we found four major haplotypes that arose due to three nonsynonymous substitutions and one 19-bp deletion leading to a frameshift. To identify three haplotypes, GmRVE8c^{hap1, 3, 4}, which are predominant in improved soybean cultivars, we developed DNA markers. Using these markers, we genotyped 129 soybean accessions, the developmental time of which had been studied in the Novosibirsk and Oryol regions. Using our data and data from SoyOmics, we found the GmRVE8c^hap3 and GmRVE8c^hap4 haplotypes to be associated with late flowering and maturity in soybean. The early-maturing haplotype GmRVE8c^hap1 is predominant in cultivars from northern regions and is likely associated with the adaptation of soybean to high latitudes. The GmRVE8c^hap4 haplotype is in complete linkage with the early-maturing allele GmTOE1^C, whereas the GmRVE8c^hap3 haplotype shows strong linkage with the late maturing allele GmTOE1^T . Furthermore, the ANOVA results indicate an interaction between GmRVE8c and E1, the major regulator of flowering in soybean. This interaction is manifested as a stronger effect of the GmRVE8c^hap3,4 haplotypes on flowering and maturity in the genetic background of the e1-as allele compared with E1. Together, these findings define a complex and intriguing locus, which may serve as a possible example of a genetically heterogeneous locus.

In different cell layers, cereal grains may accumulate various economically important polyphenols such as colored anthocyanins and melanins and colorless proanthocyanidins. To effectively create new cultivars with different combinations of these compounds, a simple, fast, and precise screening method is required. Here, a histochemical assay that includes a combination of hot ethanolic, acidic, alkaline, and ammoniacal silver treatments of grain cryosections followed by microscopy was successfully applied to distinguish these substances in cereal grains. Barley lines previously characterized chemically for the presence of anthocyanins, proanthocyanidins, and melanins in grains were used as a model. In black barley grains, this approach allowed to visually distinguish insoluble melanins that do not react to a pH change from anthocyanins, which can be insoluble or soluble but always react to changing pH. For the first time, ammoniacal silver staining commonly used for melanin identification in human and animal tissues was adapted for melanin identification in plant tissues. Along with melanins, this reagent stains other polyphenols thereby helping to detect colorless polyphenols including proanthocyanidins in the testa of barley grains as confirmed by p-dimethylaminocinnamaldehyde (DMACA) staining. The applicability of this assay to polyphenol profiling was demonstrated not only in the barley grain but also in wheat and common vetch grains. The proposed histochemical assay allows rapid polyphenol screening using a single grain, making it a practical and efficient alternative to time-consuming chromatographic methods for preliminary selection from large sample sets prior to detailed quantitative and qualitative chemical analysis.

In plants, the regulation of transgene transcription is typically achieved using chemical agents. A  safe alternative to chemically induced systems may be optogenetic systems. The BphP1-QPAS1 system has distinct advantages over other optogenetic systems, as it is activated by near-infrared (NIR, 780 nm) light, which is beyond the spectrum of plant photoreceptors. This system is based on the use of a split transcription factor (TF), consisting of the DNA-binding and dimerization domains of the yeast TF Gal4, fused to the QPAS1 component, along with the transactivation domain VP16 fused to BphP1. Under NIR light, BphP1 interacts with QPAS1, leading to the formation of the functional TF Gal4-VP16. A primary obstacle to using optogenetic systems in plants is their undesired activation under white light, which is vital for normal plant growth. A potential solution to this issue is temporarily removing one component of the split TF from the nucleus under white light. We modified the BphP1-QPAS1 system to activate reporter gene expression in Nicotiana benthamiana leaves using NIR light. We combined BphP1-QPAS1 with several variants of LOV domain-containing proteins activated by blue light (460–480  nm). The best results were achieved by combining the BphP1-QPAS1 system with the AsLOV2 domain, which carries the degron sequence RRRG at the C-terminal Jα helix and initiates the degradation of the chimeric protein NES-Gal4-QPAS1-AsLOV2-RRRG under white light. This modification induced the BphP1-QPAS1 system in tobacco leaves only under NIR light, but not in the dark or under white light. We believe that, in the future, the BphP1-QPAS1 system could be applied to enhance plant resistance to adverse environmental conditions, pests, and viral diseases.

Studying the molecular mechanisms underlying autism spectrum disorders (ASD) requires cellular models capable of capturing cis-regulatory effects and allele-specific gene expression. In this study, we present an approach for generating induced pluripotent stem cells (iPSCs) modified using an adenine base editor (ABE) to introduce synonymous single-nucleotide substitutions in the AUTS2 gene – a candidate involved in ASD pathogenesis. These substitutions serve as allele-specific markers, enabling the tracking of expression differences between normal and rearranged alleles in a cis-regulatory context. We developed a high-efficiency strategy for genotyping clones using amplicon-based next-generation sequencing (NGS). Analysis of over 100 subclones demonstrated that this approach surpasses Sanger sequencing in scalability, sensitivity, and cost-effectiveness. We selected clones with targeted heterozygous substitutions, assessed mosaicism levels, and performed phasing with germline heterozygous variants to confirm monoclonal origin and identify the allele carrying the chromosomal rearrangement. The resulting iPSC lines mark distinct AUTS2 alleles, providing a foundation for analyzing the impact of cis-regulatory elements on gene expression across different cell types. Our findings highlight the practical value of base editors and targeted NGS genotyping in creating cellular models with single-nucleotide substitutions for both basic and applied research.

Gene mutations and altered epigenetic regulation of gene expression are characteristic features of malignant neoplasms. Combinations of these abnormalities form molecular features of individual tumors. In the large-scale Dependency Map (DepMap) project, the broad panels of human tumor cell lines are being tested for sensitivity to single gene inactivation. Using DepMap data, we have previously identified a set of genes termed supertargets, the deletion of which significantly reduced the survival of cells of a particular tissue origin while minimally impairing the unrelated cell lines. In the present study, we determined the factors of viability (inhibition of proliferation or death) of cell lines in which the supertarget genes have been deleted. We found that, in 79 % of cases, the reduced survival may be caused by epigenetic changes of gene expression. In the remaining 21 % of cases, it is associated with altered gene structure. Three groups containing different types of gene expression alterations can be distinguished. In the first group, the reduced cell survival correlated with a higher expression of the supertarget gene (e.g., SOX10 and HNF1B). In the second group, a gene different from the deleted supertarget was overexpressed (gene pairs: FOXA1 and SPDEF, TP63 and SERPINB13, etc.). The third group was characterized by correlations between low expression of a certain gene and tumor cell sensitivity (e.g., FAM126A and FAM126B, SMARCA2 and SMARCA4). The genetic changes included GOF mutations (KRAS, BRAF genes, etc.), LOF mutations (STAG1, SMARCA2 genes, etc.), gene fusions (BCR-ABL1, PAX3-FOXO1, etc.), and amplification (CPM, BEST3, etc.). Therefore, many different molecular mechanisms act as predictors of tumor cell response to inhibition of supertarget genes.

In the process of adaptation to cold in humans, genes belonging to the thyroid system signaling pathways that regulate thermogenesis, energy expenditure, and metabolic rearrangements are implicated. One such gene is the THRB gene, which encodes the nuclear receptor TRβ, with which the thyroid hormone triiodothyronine (T3) interacts. The activity of thermogenin UCP1 is influenced by the concentration of TRβ-T3 complexes, which serve to uncouple oxidative phosphorylation in mitochondria, thereby enhancing heat production. Consequently, thyroid hormone receptors have been demonstrated to play a significant role in adaptive thermogenesis. In the present study, we conducted a comprehensive analysis of published data on the THRB gene polymorphism in Siberian indigenous populations, with the objective of identifying potential associations between polymorphism variants and adaptation to cold. The analysis of exon and adjacent noncoding regions of the THRB gene revealed a single nucleotide substitution in the protein-coding region (synonymous substitution in the locus rs3752874). All other nucleotide substitutions were detected primarily in 3’-untranslated regions and introns. Analysis of the THRB haplotype distribution revealed two Koryak-specific haplotypes characterized by the rs762175401-A substitution. The results of population screening demonstrated that this substitution is prevalent among the Koryak population, with a frequency of 13.8 %, and is also present in the Siberian Eskimo population. However, in other global populations, the frequency of the rs762175401-A substitution does not exceed 0.05 % (in the Japanese and Koreans) or has even lower values (less than 0.02 %). The analysis of the nucleotide sequence of the THRB gene indicates that the rs762175401 locus is situated in the 3’-untranslated region at position +2 from the terminating codon. It is plausible that this substitution may have led to alterations in translation termination efficiency. In the case of enhanced termination efficiency, it is conceivable that it contributed to an elevated rate of protein synthesis, thereby resulting in an increase in the concentration of TRβ-T3 complexes. The higher frequency of the rs762175401-A variant in the Koryak and Eskimo populations, representing the oldest populations of Northeastern Siberia, is assumed to be due to long-term adaptation of these populations to cold.

Currently, identifying biomarkers that can reliably predict the risk of developing severe COVID-19, potentially leading to fatal outcomes, remains a critical challenge. Studying the pathogenetic mechanisms underlying the progression from moderate to severe disease through blood transcriptome analysis enables the identification of differentially expressed genes (DEGs), which may serve as potential prognostic biomarkers of disease severity and as novel therapeutic targets for managing COVID-19 complications. In this review, we have summarized and analyzed studies that compared gene expression profiles between moderate and severe COVID-19 cases using bulk RNA sequencing of blood cell samples. Based on the results of five studies, five commonly and significantly differentially expressed genes were identified (CD177, PPARG, PCOLCE2, SLC51A and ADAMTS2), and their potential roles in the progression to severe COVID-19 are discussed. Functional enrichment analysis was performed, and shared pathways associated with severe COVID-19 were identified, including neutrophil degranulation, interleukin signaling, collagen biosynthesis, and suppression of adaptive and NK cell-mediated immune responses. Additionally, single-cell RNA sequencing (scRNA-seq) studies were reviewed, comparing moderate and severe cases, supporting some of the bulk RNA-seq findings. Due to the limited overlap of data in the reviewed articles, one section of this review focuses on the study designs, including analytical tools, sample collection protocols, and criteria used to define comparison groups. Transcriptomic analysis of the COVID-19 severe form reveals both cellular and molecular mechanisms of the immune response, the dysregulation of which can lead to the development of severe manifestations. RNA-markers seem to be promising predictors of the severity of COVID-19. At the same time, other omics technologies can fill in the gaps in understanding the characteristics of severe COVID-19 and identify mechanisms of disease progression to develop approaches for COVID-19 prevention and treatment.

Wild bird species contribute significantly to the rapid geographic dissemination of tick-borne encephalitis viruses (TBEV) and West Nile virus (WNV), facilitating the establishment of new natural foci of these orthoflaviviruses. However, the impact of TBEV and WNV population variability on shaping these foci, as well as the potential emergence of new human-pathogenic viral variants, remain underexplored. This study aimed to assess the genetic heterogeneity of TBEV (Siberian and Far Eastern genotypes) and WNV, isolated simultaneously from the tissues of a single garden reed warbler (Acrocephalus dumetorum) collected in the suburbs of Tomsk. The methods of viral strain isolation on various cell cultures were used in combination with a whole-genome analysis of isolates through traditional and high-throughput sequencing (NGS) methods. Consensus full-genome nucleotide sequences of the viruses were obtained by Sanger sequencing and compared with those obtained by NGS, with single nucleotide substitutions (single nucleotide variants, SNVs) accounting for 2 % or higher within the population under study. Our findings revealed single nucleotide polymorphisms (SNPs) associated with both synonymous and non-synonymous nucleotide substitutions, primarily located within the non-structural protein genes of TBEV and WNV. Notably, recombination events were not detected in the genomes of isolated orthoflaviviruses. The WNV isolate, Tomsk/bird/2006/A4, and the TBEV isolates, PT12 and PT122, obtained from A. dumetorum, exhibited heterogeneous viral populations, with SNVs ranging in frequency from 1.75 to 19.88 % for WNV and from 2.08 to 23.73 % for TBEV. Most identified SNPs shared similar nucleotide substitutions in the genomes of already known strains of TBEV and WNV, suggesting that these SNVs could play a crucial role in viral adaptation and underscore the genetic and phenotypic diversity of these viruses in nature.

Picobirnaviruses (PBVs), members of the Picobirnaviridae family, are found in a wide range of hosts, including eukaryotes (both higher and lower), fungi, and bacteria. However, scientists are unsure about their “true master” or primary host. While often found in animals, including cases of gastroenteritis, they are also detected in environmental samples and have shown genetic links to bacterial and fungal viruses. The lack of a reliable cell culture or animal model for PBV propagation further complicates determining their host specificity. Due to the discovery of prokaryotic regions (motifs) in segments of the PBV genome, it was suggested that their hosts are prokaryotic. However, even this discovery did not pin one specific host to PBVs; since then PBV-like genomes not characteristic of the studied PBV strains, with a mitochondrial genetic code characteristic of lower eukaryotes (molds and invertebrates), were discovered. And recently, a new version of the origin of PBVs from vertebrate viruses and fungi has appeared, denying their phage nature. To understand the nature of genetically diverse PBV strains detected in different organisms, researchers were guided by information about the presence of motifs specific to the viral family in the genome, the genetic code used, and the method of distribution. Recent research suggests that PBVs, previously thought to have a vertebrate origin, may have also evolved from fungal sources denying their phage nature. Some PBV-like sequences have been found to utilize the fungal mitochondrial genetic code, indicating a possible fungal origin or a close relationship with fungal viruses like mitoviruses. This discovery challenges the previously held view of PBVs as exclusively vertebrate viruses and suggests a more complex evolutionary history. The information available today inspires confidence in the imminent conclusion of the ongoing discussion about the possible PBV hosts. In particular, a hypothesis has recently emerged demonstrating a possible mechanism for the replacement of the genetic code in RNA viruses, which makes it possible to explain the origin of PBV forms with the mitochondrial genetic code capable of reproduction in cells of lower eukaryotes using the example of phages. However, an evolutionarily deterministic model demonstrating the path of PBV formation with the genetic code of mold and invertebrate cells has not yet been presented. According to the authors of this review, this evolutionary path is due to the endosymbiotic relationships between the putative PBV hosts, contributing to the horizontal virus spread. The purpose of this review article is to attempt to describe a possible path of formation from the ancestral PBV form and its derived evolutionary forms, some of which inherited a genome with a prokaryotic motif and a standard genetic code, while others acquired a non-standard form of the genome with the code of lower eukaryotes. This review article focuses on the leading role of horizontal transmission in the formation of non-standard intermediate PBV forms.

Significant gene order diversity of mitochondrial (mt) genomes of invertebrates is peculiar to subphylum Crustacea, and to order Amphipoda in particular. Amphipods from Lake Baikal are also known as a group with unique gene orders in their mt genomes. To estimate the diversity of protein-coding gene orders (GOs) in amphipods, a comparative analysis of gene rearrangements in the mt genomes of Baikal and non-Baikal species was performed. In some cases, gene rearrangement data and the history of gene relocation in different taxonomic groups can also supplement the results of phylogenetic inferences. Among the thirteen mt genomes of Baikal species sequenced in previous studies, four gene order patterns were identified, and fourteen gene order patterns for 114 mt genomes of non-Baikal species were observed. The type and number of rearrangement steps (from 1 to 3) required to transition from one order to another and the number of mt genes rearranged in each GO (from 1 to 5) were also defined. Baikalian amphipods belong to two lineages (I and II) according to molecular data which reveal their origin from two independent introductions of ancestral species into the lake. All cases of mt gene order rearrangements have been detected in species from the first lineage, whereas the mt gene order in the second lineage is conserved in all species studied and corresponds to the Pancrustacean pattern (PanGO). PanGO has been determined as the ancestral gene order for both Baikalian amphipod lineages. The possible mechanisms of mt gene order rearrangements such as a complete or partial duplication of mt genome and subsequent random deletions are discussed in our study. It is supposed that increased mutation rate, weakening of stabilizing selection and other specific factors may influence the probability of emergence and fixation of different GOs in mt genomes of Baikalian amphipods. 

Iris is a cosmopolitan genus comprising 200 to 340 species distributed throughout the Northern Hemisphere. Although Iris is the most diverse group in the family Iridaceae, there are many uncertainties regarding its taxonomic composition and systematics. The aim of this study was to search for taxonomically significant morphological characters of the generative and vegetative spheres and molecular markers with subsequent assessment of their informativeness in identifying phylogenetic relationships and compliance with the most relevant modern classification systems of the genus Iris. As a result of constructing the structure of variability of morphometric parameters of 11 species, 10 taxonomic indicators were identified that were common to the analyzed taxa and were characterized by relatively low total and coordinated variability: length and width of the outer perianth lobes, length and width of the inner perianth lobes, length of the filament, anther and pistil, fruit width, as well as seed length and width. Nucleotide sequences of trnL-trnF fragments of chloroplast DNA were established for 13 samples of four species of wild flora of the Republic of Bashkortostan and the Orenburg Region: Iris pumila L., I. scariosa Willd. ex Link., I. pseudacorus L., I. sibirica L. The obtained sequences were used to construct a phylogenetic tree together with trnL-trnF sequences of seven more iris species extracted from the database. The tree contained five clusters: (1) I. pumila, I. scariosa; (2) I. pseudacorus, I. setosa Pall. ex Link; (3) I. lactea Pall.; (4) I. sibirica, I. sanguinea Hornem.; (5) I. spuria L., I. xanthospuria Mathew & Baytop., I. foetidissima L., I. sintenisii Janka. By the composition of their species, the identified clusters almost completely coincided with the clusters found during the morphological analysis. To confirm the obtained results, a phylogenetic analysis of the species of interest was performed on two more chloroplast sequences available in the database: matK and trnS-trnG. Clustering of the studied species on trnS-trnG and matK completely coincided with clustering on trnL-trnF. Thus, we can state that the morphological features identified for the Iris generic complex work in the taxonomic direction. The analysis also showed that I. scariosa from natural populations of the Republic of Bashkortostan and the Orenburg Region were identified correctly.