Recently, genome-editing technologies have  become more efficient and accessible. The discovery of nucleases for directional genome editing (CRISPR/Cas9, TALEN, ZFNs) significantly accelerated and simplified the production of mice with targeted gene editing in the genome. Until last time, the CRISPR/Cas9 system noticeably simplified the preparation of knockout or transgenic mice. CRISPR/Cas9 technology was successfully applied for gene knockout and knock-in, generation of large deletions or directed insertions in targeted genome regions in embryonic stem cells (ESCs).When injected into blastocysts, such  modified ESCs are able to generate chimeras producing gametes with an identical genotype with ESC. Thus, it can identify animals with modified genomes. More recently, CRISPR/Cas9 technology was successfully applied to mouse zygotes and the birth of genetic modified mice was observed, i. e., the time required for generating genome-modified animals decreased significantly. The CRISPR/Cas9 system allows making gene knockout, large deletions or directed insertions into the target region of the genome by cytoplasm or pronuclear microinjection into zygotes. In addition, this is faster and simpler than similar work with mouse ESCs. Meanwhile, methods of manipulation with early embryos and their transplantation to surrogate mothers may be somewhat tricky. Therefore, it is important to use modern technologies for directional genome editing and perfect mastery in the embryological technics. In this article, we describe the protocols of microinjection into the pronucleus or cytoplasm of zygotes and injection of embryonic stem cells into the blastocyst cavity. We also describe embryological methods, such as superovulation, preparation of early stage  embryos,  surgical operation, production of foster mice. In addition, we describe the assembly and necessary components for the isoflurane anesthetic apparatus and isoflurane anesthesia.

Despite the multitude of anticancer cytostatic drugs available to oncologists today, most of such drugs have serious side effects that may preclude their use in some groups of patients. Hence, selective induction of apoptosis in cancer but not normal cells remains an attractive goal of molecular medicine. Lactaptin, a proteolytic fragment of the human milk kappa-casein, has been previously identified as a protein displaying potent killing of cancer cells in vitro. Its recombinant analog (RL2) produced in E. coli has been shown to delay solid tumor growth in vivo. Given that lactaptin is of human origin and is not immunogenic, it can be administered to patients multiple times without running the risk of immune response that could dampen the therapy efficacy. In the present study, we demonstrate that the combination of RL2 and cyclophosphamide treatments has an additive therapeutic effect against hepatoma tumor in immunocompetent mice. We asked whether production of lactaptin in human rather than bacterial cells would result in a protein with increased cytotoxic activity. Using lentiviral vector pCDH as a backbone, two constructs, pEL1 and pEL2, encoding secreted forms of lactaptin that differ in their signal sequences were created. Lactaptin expression in human cell lines was confirmed using Western-blot analysis, whereas ELISA was used for quantification of secreted lactaptin. Next, we measured the cytotoxic effects of the media conditioned by pEL1-transfected HEK293T cells, as assayed against the panel of three human cancer cell lines: MDA-MB-231 (adenocarcinoma), PC3 (prostate cancer), and T98G (glioblastoma). We show that EL1-derived lactaptin is at least 100-fold more cytotoxic than RL2. Taken together, our results provide an opportunity for developing armored immune cells as an “off-the-shelf” platform for targeted delivery of lactaptin to cancer cells. 

New technologies determining the DNA sequence of nucleotides have led to the discovery of hundreds of thousands of mononucleotide polymorphic markers, some of which are associated with breeding quality of animals. Developed on the basis of these achievements, genomic selection has produced a revolutionary shift in the poultry industry. The developed molecular marker system provides a unique opportunity to significantly improve the accuracy of estimated breeding values to manage genetic variability, to reduce the interval between generations, and accelerate genetic progress. Genomic breeding in the poultry industry has a number of differences from similar technology used in agricultural mammalian species. The existence of two categories of chromosomes (macro- and microchromosomes) with different rates of recombination, the preferred genomic females evaluation, rapid change of generations make their own features. Technology is introduced rapidly in various sectors of the poultry industry, including broiler production, and is used by major poultry companies. The individual steps of genomic selection need to be improved, and the continuous improvement of technology can be attained though its use in practical and scientific. The improvement of the separate stages of genome selection will be helped by perfection of registration and mathematical treatment of the phenotypical and molecular database, imputation and estimations of linkage disequilibrium.

Pig is the most promising species for transplantation of organs and cells into humans, although implementation of xenotransplantation in clinical practice has been hindered by the risk of infecting the recipient with zoonotic infectious diseases. Porcine endogenous retroviruses (PERV) are capable of incorporating copies of DNA into the genome of a host cell. Based on the nucleotide sequence of the envelope gene (env), three main types of pig retrovirus, PERV-A, PERV-B and PERV-C, have been recognized, with PERV-A and PERV-B having the capability of infecting human cell lines in vitro. Selection for animals with low copy number of retroviruses in the genome using simple phenotypic indications is required for the widespread implementation of xenotransplantation. The objective of this study was to evaluate the correlation between PERV-A env gene copy number and hematological parameters, gender and coat  color in miniature pigs of the Institute of Cytology and Genetics (ICG) SB RAS. Reference values for eighteen blood parameters of miniature pigs were determined, including white blood cell count (WBC), red blood cell count (RBC), platelet count (PLT), absolute (LYM#) and relative (LYM%)  lymphocyte counts, absolute (MID#) and relative (MID%) monocyte, basophil and eosinophil counts, absolute (GRA#) and relative (GRA%) granulocyte counts, hematocrit (HCT) and thrombocrit (PCT), mean cell volume (MCV) and mean platelet volume (MPV). Males had significantly higher reference values for WBC, MID#, GRA# and red cell distribution width (RDW-CV) as compared to females. The mean corpuscular hemoglobin  concentration (MCHC) and platelet distribution width (PDW-CV) were significantly higher in female animals. No correlation between PERV-A env gene copy number and the coat color of  animals was detected, suggesting that retroviral insertion sites and genes that determine the coat color of miniature pigs, namely KIT (chromosome 8) and MC1R (chromosome 6), are either located far apart on same chromosome or on different chromosomes. The copy number of PERV-A env gene in males was lower than in females. Presence of multiple copies of PERV-A on the X-chromosome is the most probable cause of such gender-related differences in miniature pigs. Thus, male miniature pigs of ICG SB RAS should be the source of material for xenotransplantation.

The spectrum of the metabolites in the dorsal region of the hippocampus and striatum was studied using the method of 1H magnetic resonance spectroscopy at experimental modeling of the posttraumatic stress disorder syndrome (PTSD) in rats. PTSD was reproduced by exposure of the cat cue to rats daily along 10 day by 10 minutes at once. The anxiety level of animals was estimated 12 days later after the end of the experimental series of stress. Based on the anxiety index, the rats were divided into 3 phenotypes. The animals with an anxiety index > 0.8 (group 1) had lower plasma corticosterone compared with rats form the control group. In animals with an anxiety index in the range 0.7–0.8 (group 2), an elevated corticosterone level was noted. The rats with an anxiety index < 0.7 (group 3) had a lower plasma corticosterone level compared with animals from the control group. Rats of group 2 were characterized by an increased level of GABA in the hippocampus compared with controls. In the remaining groups, the percentages of GABA in the hippocampus and striatum did not differ significantly from the control. The distribution of NAA differed form that of GABA. The highest level of NAA was found in the striatum for rats from group 1, whereas NAA in animals form groups 1 or 3 did not differ from the control. The NAA level in the hippocampus was similar between all groups, including the control. The results obtained indicate that multiple exposures to psychological stress associated with the sense of proximity of a natural enemy in some animals cause an anxiolytic reaction. These animals are characterized by a stable corticosterone level and a stable level of neurometabolites in the studied structures of the brain. For rats with the highest level of anxiety, a lowered level of corticosterone with a constant level of neurometabolites in the hippocampus and striatum is characteristic. And only in rats with an intermediate level of anxiety, synchronization was observed between the increase in plasma corticosterone and the increase in hippocampal GABA content. The results obtained are in good agreement with the ideas of the protective action of glucocorticoids under PTSD manifested in  restraining violations of the psycho-physiological status. The mate rials allow the neurobiological mechanisms of the protective action of glucocorticoids to be detailed.

The behavioral and neuronal responses of the males of two closely related species of mice (Mus musculus  wagneri, M. spicilegus) to the urine odors of estrus con- and heterospecific females were studied. In two-choice odor tests males significantly longer investigated odor of conspecific females in comparison with heterospecific ones. To investigate neuronal activation in the main and accessory olfactory bulbs (MOB and AOB), one of the methods of functional tomography – manganese-enhanced MRI ( ME-MRI) – was used. There was a significant increase in Mn2+ accumulation in the dorsal section of the posterior part of the MOB in male M. m. wagneri and M. spicilegus exposed to odor of conspecific females compared with the control group males (odor not exposed). There was a local significant increase in manganese accumulation in the dorsal region of the anterior part of the MOB in the case of the exposure of odor of a heterospecific female. The exposure of heptanone-2 to mice resulted not only in an increase in Mn2+ accumulation in certain zones, but also in a significant decrease in the accumulation of Mn2+ in the rest of the olfactory bulbs. A significant increase in the accumulation of MRI contrast in AOB was observed in males only in the case of female urine-conspecific odor exposure. The results support the previously stated assumption of a significant difference in chemical communication systems in two species of mice. A comparison of these results and results obtained previously demonstrated the absence of any differences in behavioral and neuronal responses to con- and heterospecific odors of the house mouse subspecies allopatric (M. m. wagneri) and sympatric (M. m. musculus) to M. spicilegus. This fact does not allow us to assume the effect of the mechanism of “reinforcement” in the process of evolution in the formation of precopulatory reproductive isolation between the sympatric species M. spicilegus and M. musculus.

Striking similarities between human and miniature pig anatomy and physiology of the cardiovascular system, skin and digestive organs have made miniature pigs a successful animal model for addressing problems of xenotransplantation. However, despite the widespread interest in this breed of animals, the distribution of water and electrolytes in the tissues of miniature pigs remains poorly understood, which led us to conduct this study. The distribution of sodium, potassium cations and water in skeletal muscles, myocardium, smooth muscle elements of the vascular system, liver and skin of mini-pigs differing in gender and behavioral response to humans has been studied. Quantitative determination of sodium and potassium cations was performed by atomic absorption spectroscopy. It was shown that skeletal and smooth muscles differ in electrolyte content. The impact of behavioral response to humans on the electrolyte level in skeletal muscle was detected. The data are confirmed by two-factor dispersion analysis. The accumulation of water in muscle samples of “tame” males as compared with cowardly animals was shown. The impact of different behavioral types on the amount of sodium in the skin was revealed. The impact of the gender of animals or the type of behavior on the sodium level in the liver was not detected. However, a significant amount of potassium was found in the liver as compared with sodium level, which is possibly associated with a high density of cellular elements. The gender and behavioral type of animals did not have an impact on the distribution of potassium and the mass fraction of water in the skin and liver. Some differences in the distribution of the main extracellular and intracellular sectors’ cations were revealed, and so was the dependence of the electrolyte content on the functional characteristics of the tissues. The data obtained demonstrate that miniature pigs can be an informative animal model for  studying the influence of genetic (gender) and phenotypic (behavioral response) aspects on the parameters of water-electrolyte homeostasis.

The GC rat strain (from the words “genetic” and “catatonia”) was created by selection for predisposition to passive-defensive reaction of catatonic freezing in response to stressing stimuli. Rats of the GC strain have previously demonstrated a number of biochemical and behavioral properties similar to those of patients with schizophrenia and depression. Prepulse inhibition (PPI) is widely explored as an important indicator, a decrease of which may be indicative of psychopathology, including schizophrenia. It has been established that the brain noradrenergic system influences the manifestation of PPI, in particular through the activation of central alpha-adrenoreceptors. Also known is the association between PPI and expression of catechol-O-methyltransferase. This study focuses on the reaction of prepulse inhibition in rats of the inbred GC strain, being considered as a hypothetical model of schizophrenia, as well as on the relation of prepulse inhibition to mRNA expression of tyrosine hydroxylase, catechol-O-methyltransferase, alpha1A- and alpha2Aadrenergic receptors in the midbrain of GC rats. For the first time, a decrease of PPI in GC rats compared with WAG rats was shown, both with a prepulse power of 75 dB and at 85 dB, which may indicate a violation of filtration of sensorimotor information into the central nervous system in GC rats. Real-time PCR showed a decrease in mRNA level of Adra1A in intact rats with genetic catatonia when compared to control WAG rats. There was observed no correlation between the expression of mRNA of the Adra1A, Adra2A, Th, and Comt genes in the midbrain and the PPI reaction in GC rats. The reduction of prepulse inhibition in GC rats indicates functional similarity of this genetic model of schizophrenic psychopathology with a prototype.

The Zbtb33 gene encodes the bimodal transcriptional repressor Kaiso, which causes epigenetic repression of genes by binding to methylated mCpG islets in the promoters of the genes. Despite the fact that Kaiso is intensively expressed in the central nervous system, its participation in the regulation of behavior is still poorly understood. Only the participation of Kaiso in the regulation of the behavioral response to emotional stress in the open field and forced swimming tests has been shown. The aim of this study is to elucidate the role that Kaiso plays in regulating daily activity, as well as the behavioral response to stimulation of nonspecific immunity. Experiments were performed on adult male mice with Zbtb33 gene knockout (KO) and animals of the C57BL/6 line (wild type, WT). All animals were 11 weeks old, weighed 26 ± 1 g and had SPF (specific pathogen free) status throughout the experiment. The animals of each genotype were divided into three weighted groups of 8 animals each. Initially, the daily dynamics of motor activity, sleep, food and water intake of intact animals was measured using the PhenoMaster software-hardware complex. The animals of each group were then injected with saline (control), 0.1 or 1.0 mg/kg of bacterial lipopolysaccharide (LPS) dissolved in saline, and again measured for their daily activity, food and water intake. Intact KO and WT mice did not differ in the average daily motor activity and sleep duration. However, intact KO mice were less active in the dark time, and also consumed less food and water as compared to intact WT animals. LPS at both doses suppressed motor activity, prolonged sleep duration and caused anorexia in mice of both genotypes. However, the effect of low dose of LPS (0.1 mg/kg) on the food and water intake was more pronounced in KO mice than in WT animals. The results shed light on the biological significance of the Kaiso gene and serve as a justification for the necessity of the normal functioning of this gene in natural populations.

OXYS rats are the model of precocious senescence. Numerous studies addressed physiology and behavior in rats of this strain during a postnatal period of their life, however, preimplantation development in OXYS rats has not yet been investigated. This study is addressing preimplantation embryonic development in OXYS rats both in vivo and in vitro. Rats of the WAG strain were used as controls. For studying the in vivo development, the embryos were collected from OXYS and WAG rats on day 5 post coitum, the stages of embryo development were estimated, the percentage of embryos at blastocyst stage and the cell numbers in these blastocysts were counted. In a special experiment, for studying in vitro development, the embryos were collected from both rat strains on day 4 post coitum and were cultured in vitro in P1 medium for 48 hours with or without supplementation with IGF-1 (200 ng/mL). Thereafter the percentage of embryos at blastocyst stage and the cell numbers in these blastocysts were counted in the same manner as for the in vivo experiment. This study reports that in vivo derived blastocysts of OXYS rats contain fewer cells on day 5 of their development than in vivo derived blastocysts of WAG rats. In vitro culture of the early preimplantation embryos in P1 medium mitigated the difference in the rate of embryo development between these two strains, the addition of IGF-1 into culture medium exerts neither negative nor positive effect on the rate of in vitro embryo development in rats of both strains.

Long-lived systems are expected to be stable, i. e. resistant to either external influences, or internal failures. Robustness of biological systems can be defined as a reciprocal value to their phenotypic plasticity expressed through a coefficient of variation (C.V.) for positively distributed phenotypic traits. Considering lifespan as phenotype, which integrates all functions of an organism, we showed that its phenotypic robustness correlates positively with life expectancy. We assessed lifespan parameters for a selection of inbred Drosophila melanogaster strains from Drosophila Genetic Reference Panel (DGRP) reared at 29 ºС. The robustness of lifespan phenotype (C.V.–1) correlated positively with estimated life expectancy for these strains. The same relation also holds for the lifespan of all DGRP strains reared at 25 ºС. Also, in agreement with previous observations, upon temperature change (decrease or increase) the survival curves scaled in time (stretched or shrunk respectively). In other words, the average lifespan decreased for flies reared at elevated temperature, but so did the standard deviation, and thus the coefficients of variation remained in the same range. From this we conclude that coefficients of variation correlate with life expectancies and account for the robustness of lifespan phenotype irrespective of accelerated aging caused by temperature.

The neuroendocrine stress-response is an effective defense mechanism against adverse influences of various nature. This reaction is universal and appears in response to stimuli that are unusual not just for living and habitat of the species, but also for each population. Here we review a progressive change of theoretical concepts, approaches and methods of research in this scientific field: beginning with the development of the stress concept by H. Selye and up to the present day. In 1982 H. Selye defined stress as a combination of stereotypical phylogenetic programmed  reactions of the organism that are caused by any strong, superstrong or extreme influences and are followed by a reorganization of the organism’s adaptive forces. The stress-causing agent was named a stressor. In the dy namics of the complex of nonspecific protectiveadaptive reactions that respond to a stressful influence aimed at cultivating the organism’s resistance to any factor, it is possible to logically identify three stages (“the Selye triad”): 1) alarm state, 2) resistance state, 3) exhaustion state. The duration and the expression of each stage can vary depending on the nature and strength of the stressor agent, the species of the animal and the physiological state of the organism. The lack of a hypothalamic-pituitary-adrenocortical system in insects was considered a proof of their inability to develop a stress reaction of the warm-blooded animals’ type. Nevertheless, since the early 1980s, enough evidence of the development of stress reaction in insects has been gathered, which emphasizes the conservative nature of the stress reaction in mammals and insects. The similarity in the neurochemical and physiological changes in invertebrates and vertebrates in response to a stressful influence indicates that the response to a stressor is a complex of ancient mechanisms preserved in evolution. Insects present unique opportunities for experimentation, which can allow us to understand the basic mechanisms of stress reactions. In insect larvae the mechanism of stress reaction has been studied in detail. In this century, the main efforts of researchers are aimed at studying the mechanisms of stress reaction in imago and genetic control of its individual links. The study of stress reaction in insects has both theoretical importance, as it demonstrates the convergence of evolutionary pathways of adaptive transformations in such distant taxa as insects and mammals, and practical importance, since the patterns of this reaction’s mechanisms can be used in modeling hereditary or acquired human diseases, in developing breeding methods for economically valuable insects and in finding ways to fight insect pests

Reduced insulin sensitivity plays an important role in the pathogenesis of type II diabetes. There are sex differences in the development of metabolic disorders. The aim of this work was to investigate the insulin signal transduction gene expression in mice of different sex and age. Male and female C57BL/6J mice were used in our studies. Gene expression was assayed by RT-PCR. It was shown that insulin sensitivity in females was higher than in males regardless of age because the level of glucose in blood plasma of females does not differ from that in males, whereas female insulin levels were lower than male insulin levels. Female glucose tolerance increased with age, and glucose tolerance was higher in females than in males at the age of 30 weeks. It was shown that sex and age affect the expressions of insulin signal transduction genes. It was shown that there are sex differences in the levels of mRNA Pik3cd in the liver, in the levels of mRNA Irs1 in the muscle, in the levels of mRNA Irs1 and Slc2a4 in adipose tissue at the age of 10 weeks, and there are sex differences in the levels of mRNA Irs2 and Pik3cd in the liver, in the levels mRNA Pik3cd and Slc2a4 in the muscle, in the levels mRNA Insr and Pik3cd in adipose tissue at the age of 30 weeks. In young animals, the expression of the genes was higher in females than in males in all tissues. In adult animals, the expression of the genes in the liver was higher in females than in males, the expression of the genes in muscle and adipose tissues were lower in females than in males. In males, the levels of mRNA Insr in the liver and muscles and mRNA Pik3cd in adipose tissue decreased with age, and the level of mRNA Pik3cd in muscles  increased with age. In females, the levels of mRNA Irs1 in muscle and mRNA Pik3cd and Slc2a4 in adipose tissue decreased with age. Thus the molecular basis of sex differences and age-related changes in insulin sensitivity may be a change in expressions of insulin signal transduction genes in the target tissues.

In recent years, the microalgae (Chlorella vulgaris) have increasingly attracted great interest as a potential source of pharmacologically active compounds. Showing anticoagulation, antioxidant and antitumor activities of Chlorella revealed its hypotensive properties. The aim of this study was to evaluate the effects of Chlorella suspension on the weight of the animals, their moving activity, and erythropoiesis. The study was performed on males and females of ICR mice. The animals from the experimental group drank only the Chlorella suspension during 3 weeks and were given standard food. Control animals drank during this period only water and had the same food. The body weight of males in the control and the experimental group with Chlorella did not change, while females in the experimental group showed an increase of body weight in a week. A similar pattern was obtained for estimation of animal body weight changes relative to food consumption. The number of red blood cells in females and males from group with Chlorella increased only after 3 weeks after the start of the experiment. Hemoglobin also increased only after 3 weeks after the start of Chlorella consumption, but only for females. All groups of animals had the same motor activity during experiment. Blood sampling resulted in a reduction of activity in control males and females as well as in males with Chlorella. The motor activity of females with Chlorella after blood sampling did not change. So, consumption of the Chlorella suspension by females leads to more effective digestion and resulted in increased body weight, improved erythropoiesis resulted in increased red blood cells and hemoglobin and also increased their resistance to acute stress. The males in the same situation increased only the erythropoiesis.

The ability to deliver particulated xenobiotics and therapeutic drugs directly from the nasal cavity to the central nervous system, bypassing the hemato-encephalic barrier, determines a high importance of investigation of factors influencing this process. It was shown that the bioavailability of solid particles is influenced by their size and surface charge. At the same time, the impact of a crystal structure (crystalline/amorphous) has been poorly investigated. In this study, using sexually mature male C57BL/6J mice, we analyzed the efficiency of the nose-to-brain transport of crystalline and amorphous manganese oxide nanoparticles. T1-weighted magnetic resonance imaging (MRI) was used to evaluate the accumulation of manganese nanoparticles in olfactory bulb (OB) and olfactory epithelium (OE). So, it has been established that amorphous particles have higher accumulation rate in OE and OB in comparison with crystalline particles after their intranasal administration. The unequal ability of amorphous and crystalline particles to overcome the mucosal layer covering the OE may be one of the possible reasons for the different nose-to-brain transport efficiency of particulated matter. Indeed, the introduction of mucolytic (dithiothreitol) 20 minutes prior to intranasal particle application did not influence the accumulation of amorphous particles in OE and OB, but enhanced the efficiency of crystalline nanoparticle entry. Data on the different intake of amorphous and crystalline nanoparticles from the nasal cavity to the brain, as well as the evidence for the key role of the mucosal layer in differentiating the penetrating power of these particles will be useful in developing approaches to assessing air pollution and optimizing the methods of inhalation therapy.

Huntington’s disease is a hereditary neurodegenerative disorder caused by CAG trinucleotide repeat expansion in the first exon of HTT gene. The mutant HTT protein has an elongated polyglutamine tract and forms aggregates in the nuclei and cytoplasm of the striatal neurons. The pathological processes occurring in the medium spiny neurons of Huntington’s disease patients lead to neurodegeneration and consequently to the death. The molecular mechanisms of the pathology development are difficult to study due to the limited material availability and late onset of the manifestation. Therefore, one of the important tasks is generation of an in vitro model system of Huntington’s disease based on human cell cultures. The new genome editing approaches, such as CRISPR/Cas9, allow us to generate isogenic cell lines that can be useful for drug screening and studying mechanisms of molecular and cellular events triggered by certain mutation on an equal genetic background. Here, we investigated the viability and proliferative rate of several mutant HEK293 cell clones with mutations in the first exon of HTT gene. The mutant clones were obtained earlier using CRISPR/Cas9 genome editing technology. We showed that mutant cells partially reproduce the pathological phenotype, that is, they have reduced proliferation activity, an increased level of apoptosis and high sensitivity to treatment with 5μM MG132 proteasome inhibitor compared to the original HEK293 Phoenix cell line. Our results indicate that the mutation in the first exon of HTT gene affects not only neurons, but also other types of cells, and HEK293 cell clones bearing the mutation can serve as in vitro model for studying some mechanisms of HTT functioning.

Among the pathogenic complex of the common wheat there is a causal agent of leaf and stem diseases: (rusts and powdery mildew), which has economic importan ce for most of the Russian regions. We propose a methodological approach for confirmation of effec tiveness of wheat genes conferring broad-spectrum resistance to different types of fungal diseases, which integrates traditional field tests and current genotyping techniques with molecular DNA markers. The pro posed approach includes the following steps: 1) evaluating of wheat genotypes using molecular DNA markers; 2) field tests for genotypes; 3) confirma tion of effective ness of broad-spectrum resistance genes by matching field scores and data from mole cular markers. A protocol has been proposed for geno typing varieties and wheat lines using markers linked to Lr16/Sr23, Lr24/  Sr24, Lr19/ Sr25, Lr26/Sr31/Yr9/Pm8, Lr37/Sr38/Yr17 and the gene Lr34(=Sr57/Yr18/Pm38) with a pleiotropic effect. Markers to the genes Lr6Ai#2/Sr6Ai#2/Pm6Ai#2 transferred from Thinopyrum intermedium into the genomes of Russian wheat varieties and markers to a new translocation from Aegi lops spel toides with a group of genes, designated as LrAsp7/SrAsp7/PmAsp7, were tested. When evaluating the contribution of the genes (including genes located on the alien translocations) to the formation of common wheat resistance to fungal diseases, it is necessary to use an extended sample, including genotypes carrying the same group of genes in a different genetic background. In addition, recommendations are given on the terms and frequency of monitoring of fungal diseases in Western Siberia, depending on the pathogen. The methodological approach proposed in the article can be used for identification and evaluation of the efficacy of the genes determining protection of wheat from fungal diseases. This approach is applicable in the investigation of genetic collections consisting of isogenic lines, sources and donors of resistance genes, as well as in the development of wheat genotypes
using marker-assisted selection.