mRNA vaccine technologies have been actively developing since the beginning of the 21st century and have received a major boost from new findings about the functioning of the immune system and the development of efficient vehicles for nucleic acid delivery. The mRNA vaccine demonstrates superior properties compared to the DNA vaccine, primarily due to accelerated mRNA vaccine development, enhanced flexibility, and the absence of integration into the genome. Artificial mRNAs have biotechnological and medical applications, including the development of antiviral and anticancer mRNA therapeutics. The effective expression of therapeutic mRNA depends upon the appropriate selection of structural elements. Along with the addition of the 5’-cap, appropriate polyadenylation, and sequence codon optimization, 5’- and 3’-untranslated regions (UTRs) play an important role in the translation efficiency of therapeutic mRNAs. In this study, new plasmids containing a novel combination of UTR pairs, namely 5’-UTR-4 and 3’-UTR AES-mtRNR1, were constructed to obtain artificial mRNAs encoding green fluorescent protein (GFP) and firefly luciferase (FLuc) with new structural elements and properties. The novel combination of the UTRs, which is described in this article for the first time, in addition to sufficient polyadenylation and pseudouridinilation of mRNA, was demonstrated to strongly increase the translation of codon-optimized sequences of reporter mRNAs. We generated lipoplexes containing the aforementioned reporter mRNAs and liposomes composed of cationic lipid 2X3 (1,26-bis(cholest-5-en-3beta-yloxycarbonylamino)-7,11,16,20-tetraazahexacosane tetrahydrochloride) and helper lipid DOPE (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine). For in vivo experiments, the liposomes were decorated with 2 % of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG₂₀₀₀). The translation efficiency of mRNAs was found to be superior for the novel UTR combination compared with HBB gene UTRs, both in vitro and in vivo. When mRNA is administered intramuscularly, the proposed combination of UTRs provides lasting expression for more than 4 days. The results demonstrated that the novel UTR pair combination could be useful in the development of artificial mRNAs with enhanced translation efficiency, potentially reducing the dose for mRNA-based therapeutics.

Transcription factors represent one of the major groups of proteins, whose suppression leads to tumor growth arrest. Different types of cancer express a specific set of transcription factors that create and maintain unique patterns of gene expression. In prostate cancer cells, one of the key transcriptional regulators is the HOXB13 (Homeobox B13) protein. HOXB13 is known to be an important regulator of embryonic development and terminal cell differentiation. HOXB13 regulates the transcription of many genes in normal and transformed prostate cells and is also capable of acting as a pioneer factor that opens chromatin in the regulatory regions of genes. However, little is known about the protein partners and functions of HOXB13 in prostate cells. In the present study, we searched for protein partners of HOXB13 by immunoaffinity purification followed by high-throughput mass spectrometric analysis (IP/LC-MS) using the PC-3 prostate cancer cell line as a model. The main partners of HOXB13 were found to be transcription factors with different types of DNA-binding domains, including the TBX3, TBX2, ZFHX4, ZFHX3, RUNX1, NFAT5 proteins. Using the DepMap resource, we have shown that one of the identified partners, the TBX3 protein is as critical for the growth and proliferation of prostate cancer cell lines in vitro as HOXB13. Analysis of individual prostate cancer cell lines revealed that knockout of both genes, HOXB13 and TBX3, leads to the death of the same lines: VCaP, LNCaP (clone FGC), PC-3 and 22Rv1. Thus, HOXB13 and TBX3 can be considered together as potential targets for the development of specific inhibitors that suppress prostate cancer cell growth.

The assessment of intraspecific variability of wheat has been relevant for years. Although most modern wheat cultivars are considered to be pure lines, the heterogeneity of varietal populations is one of the mechanisms for maintaining population homeostasis. It is possible that the high evolutionary stability of constitutive heterochromatin and its stable distribution within chromosomes will allow us to use karyological analysis not only for studying the genesis and taxonomy of Triticum L., but also for studying the intraspecific diversity of wheat. In this regard, a classification of 87 Russian cultivars of common wheat differing in breeding status (landraces and modern cultivars) and growth habit (spring and winter) was carried out using two alternative approaches for assessing karyograms. The first approach uses the qualitative assessment of karyograms based on the distribution of C-bands on chromosomes. We also proposed that quantification of karyograms based on the size of C-bands would make the classification of cultivars more adequate. The variability, informative value and resolution of diagnostic features, trends in grouping cultivars, and their associations with the breeding status and growth habit were studied. A high potential of karyotyping with C-banding in discriminating modern cultivars by growth habit, as well as in separating winter cultivars from landraces has been revealed. In terms of the tested karyological features, the homogeneity of modern cultivars was higher than that of local cultivars, and the homogeneity of winter wheat was higher than that of spring wheat. The obtained classification reflects the preservation of high similarity in the karyograms of modern spring cultivars and landraces, as well as the low distinguishability between the karyograms of landraces differing in growth habit. A comparative analysis of the classifications of 20 cultivars using C-banding and SNP genotyping (3,126 polymorphic markers) suggests that studying the karyotypic variability allows us to infer a more accurate differentiation of wheat varietal populations based on the breeding status than using SNP markers that detect genetic variability, especially when the number of diagnostic features is limited.

The duration of the vegetation period significantly impacts yield formation and is one of the important characteristics of spring common wheat (Triticum aestivum L.) varieties. The primary developmental phases influencing the vegetation period include the time from seedling emergence to heading and from heading to maturity. To identify genes and loci associated with these traits under long-day conditions typical of Western Siberia and to assess their impact on yield components, we conducted QTL mapping followed by an evaluation of yield-related traits in lines carrying different alleles of key heading time genes. For mapping, we used an F2 population derived from a cross between the varieties Obskaya 2 and Tulun 15, which contrast in their heading and maturity times. QTL analysis identified a novel locus on the long arm of chromosome 7B associated with maturity time, as well as two loci on chromosome 2D and the short arm of chromosome 7B associated with heading time. Gene analysis within these loci revealed candidate genes for the “seedling-maturity” trait, with expression patterns corresponding to the known maturity time regulator NAM-1. The localization of loci for the “seedling-to-heading” trait suggested their correspondence to the well-known genes Ppd-D1 and Vrn-B3. Analysis of progeny carrying the Ppd-D1a and Vrn-B3a allele combination demonstrated that Ppd-D1a had a stronger effect on heading time than Vrn-B3a, and their combined presence resulted in the earliest heading – on average, five days earlier than in lines with the Ppd-D1b and vrn-B3 alleles. Evaluation of yield-related traits (number and weight of grains per main spike and per plant, and 1,000-grain weight) indicated that Ppd-D1 was significantly associated with all traits, with the Ppd-D1a allele generally exerting a negative effect on yield. In contrast, Vrn-B3 had a comparatively smaller effect on yield traits than Ppd-D1.

One of the sources of resistance to leaf and stem rust pathogens for bread wheat is the Australian spring triticale cultivar Satu, which carries highly effective linked SrSatu/LrSatu genes localized on chromosome 3R. However, they are little used in the practical breeding of Triticum aestivum L. The main reason for that is a low level of knowledge regarding the 3R(3D) chromosomal substitution. This paper presents the results of a comparative study of the agronomic value of near-isogenic spring bread wheat siblings, L16 and L17 = Satu/Saratovskaya 70//Saratovskaya 74/3/ Saratovskaya 74, differing by presence (L16 (3R(3D))) or absence (L17 (3D3D)) of chromosome 3R from Satu in 2023– 2024. The 3R(3D) chromosomal substitution in L16 was detected by cytogenetic analysis combining GISH with labeled Secale cereale genomic DNA and FISH with probes pSc119.2, pAs1. Line L16 is highly resistant to Puccinia triticina and P. graminis, including the Ug99 race. PCR analysis with DNA markers of Sr genes revealed the non-identity of the resistance gene in L16 to Sr genes: Sr2, Sr24, Sr25, Sr28, Sr31, Sr32, Sr36, Sr38, Sr39, Sr47 and Sr57. L16 was inferior to both L17 and the standard cultivar Saratovskaya 76 in terms of 1,000-grain weight. An analysis of productivity elements of the main ear revealed that the 3R(3D) substitution in L16 significantly reduced the length of the ear, increased the density of the ear and did not significantly affect the number of spikelets and the number of grains per ear and the grain weight per ear. The grain protein content in L16 did not significantly differ from its L17 siblings or Saratovskaya 76. Similarly, there were no significant differences in gluten content. However, gluten in L16 was weaker in comparison with line L17 and Saratovskaya 76. According to the complex trait of SDS sedimentation, L16 was inferior to L17, but did not significantly differ from the standard cultivar. According to the alveograph, L16 had significantly lower dough elasticity and flour strength, but in comparison with the standard cultivar, the decrease in flour strength was not significant. L16 showed a higher bread volume than Saratovskaya 76, but did not significantly differ from its L17 sibling. There was no difference in porosity for all three samples. In general, in terms of the complex of agronomically valuable traits, the spring bread wheat line L16 (3R(3D)) requires further work to improve its breeding value.

The composition of wheat grain plays a key role in determining its nutritional value. In this work, a collection of 133 durum wheat varieties and breeding lines was assessed for the content of macroelements (Ca, Mg, K), microelements (Cu, Mn, Zn, Fe, Na) and toxic metals (Pb, Cd, and Cr) in grain under the environmental conditions of Samara and Novosibirsk regions in 2023. The results showed a wide range of variations in the concentration of all elements depending on genotypic differences between the samples as well as the growing region. Ca and Mg contents in the varieties grown in Samara region showed a significant excess of 3.1- and 1.5-fold, respectively. Zn, Pb, and Cr content in the varieties cultivated in Novosibirsk turned out to be two times as high. Statistical analysis of element concentrations in the varieties of different origin indicates that Russian breeding lines significantly outperform Russian cultivars in Mg content, while being inferior in K, Cu, and Mn. Compared to Russian cultivars and breeding lines, foreign varieties demonstrated higher contents of K and heavy metals Cd and Cr. Correlation analysis using mean values of indicators for two environments showed highly significant (p < 0.001) positive relationships between the content of microelements Fe/Mn (r2 = 0.69), Fe/Zn (r2 = 0.49), and Zn/Mn (r2 = 0.46), which suggests a feasibility of selecting genotypes for several elements at once. Multivariate statistics divided the durum wheat collection into two groups, one of them including Russian cultivars and breeding lines as well as some foreign genotypes. A separate cluster included seven Russian breeding lines placed at a distance from the other varieties, which suggested their potential differences at the genetic level. Comparing these lines with respect to mineral composition showed that they were, on average, characterized by higher Mg, K, Zn, and Fe contents. The data obtained in this study can be used for genetic research and breeding to improve the grain mineral composition of the modern durum wheat varieties.

Little ground squirrel Spermophilus pygmaeus (Pallas, 1778) is a polytypic species of significant interest for the study of taxonomic diversity, genetic structure, gene flow and genetic diversity. Despite the long history of study, the taxonomy of representatives of the genus Spermophilus in the North Caucasus remains poorly developed. Among the unresolved issues are the phylogenetic relationships between the “mountain” and “plain” ground squirrels of the North Caucasus. An equally important aspect of the work is the study of the genetic diversity of little ground squirrel, given that the species is considered an integral component of steppe and desert ecosystems, providing their most important biocenotic functions. Based on the analysis of the 840 bp mtDNA cytochrome b gene fragment, new data on the genetic variability of S. pygmaeus from the eastern extremity of the Western Caucasus were obtained. Unlike previous studies that showed the so-called mountain ground squirrel to inhabit the Caucasus Mountains, this work identified two haplogroups of S. pygmaeus in the studied areas at an altitude of 1,400–1,700 m above sea level, one of which is close to the lowland (East Caucasian) and the other to the mountain (Central Caucasian) groups of the little ground squirrel. The genetic distance between the two haplogroups was 1.54 %. The different evolutionary ages of the three identified groups of S. pygmaeus in the North Caucasus (A1, A2, and B) are most likely associated with the multi-stage settlement of the studied area by the little ground squirrel. The results of molecular dating suggest that the western haplogroup penetrated as a continuous strip into the Central, Eastern Caucasus and the eastern extremity of the Western Caucasus through the Stavropol Upland and the Caspian Lowland less than 400 thousand years ago. As a result of the first wave of dispersal of the ground squirrel from the Russian Plain, the species became established in the eastern extremity of the Western Caucasus in the area of the village of Khasaut, and in the Eastern Caucasus – in the north of the Nogai Steppe (Sukhokumsk) and in the southern outskirts of the Caspian Lowland (Kar-Kar 1 Valley). The younger age of haplogroup A2 (less than 300 thousand years), also originating from the Eastern Caucasus (Khumtop, Zelenomorsk, Lvovsky 13, Kar-Kar 2), is most likely due to the re-colonization of the Caspian lowland by the ground squirrel, which was regularly flooded by the Caspian Sea in historic times. The absence of a continuous forest belt in the Central Caucasus, in particular in the Kabardino-Balkarian Republic, allowed S. pygmaeus to penetrate into the mountains later, less than 200 thousand years ago, through three gorges: Cherek, Baksan and Malkinsky. It is more likely that the species penetrated into the subalpics of the Western Caucasus (Khurzuk and Uchkulan) from the Central Caucasus, as evidenced by the same evolutionary age of animals of the Western (Uchkulan, Khurzuk) and Central Caucasus. Regarding the taxonomic status of the Caucasian mountain ground squirrel, we consider it premature to draw any conclusions, since not all areas of the Caucasus were covered by research.

The ability of trehalose to improve metabolic parameters in mice with experimental obesity has been shown to depend on the type of obesity model. In db/db mice, it reduced body weight, insulin, blood glucose, and cholesterol levels. In mice with obesity induced by high-fat dietary intake, it had no effect on body weight but reduced blood insulin levels with compensatory upregulation of insulin signaling gene expression. We studied the effect of trehalose on overweight and metabolic parameters in C57BL/6 inbred mice with obesity induced by a high carbohydrate-fat diet, the “cafeteria diet”. The cafeteria diet consisted of free access to water, standard chow, fatty foods (lard), and carbohydrates (biscuits) for 18 weeks. All mice were then randomly divided into four groups for four weeks of treatment: (1) water drinking, (2) drinking 3 % trehalose, (3) cafeteria diet and drinking water, (4) cafeteria diet and drinking 3 % trehalose. Alterations in body mass, food intake, fluid intake, dietary calories, blood biochemical parameters (glucose, triglyceride, cholesterol, HDL, ALT, creatinine levels), expression of carbohydrate metabolism (Slc2a2, Insr) and autophagy (Atg8, Becn1, Park2) genes in the liver were studied. The cafeteria diet obesity model was accompanied by some signs of metabolic syndrome as it induced an increase in body weight (by 25 %), calorie intake (by 25 %), blood levels of glucose (by 35 %), cholesterol (by 66 %), and triglycerides (by 23 %) in mice. Trehalose had little effect on control mice, causing a decrease in standard food intake and an increase in dietary caloric intake by the number of calories from trehalose itself. In obese mice, trehalose increased total caloric intake and biscuit consumption but had no substantial effect on body weight gain, blood metabolic parameters, or expression of liver genes regulating glucose transport (Slc2a2), insulin sensitivity (Insr), and autophagy processes (Atg8, Becn1, Park2). Since the cafeteria diet is the most adequate model of alimentary obesity development in humans, our results question the use of trehalose to correct the dietary type of obesity in humans.

Recent genome-wide association studies have identified a link between the RAB29 gene and Parkinson’s disease (PD). The Rab29 protein encoded by RAB29 regulates leucine-rich repeat kinase 2 (LRRK2). Mutations in the LRRK2 gene increase its kinase activity and contribute to autosomal dominant forms of PD. Previous research has shown that altered LRRK2 kinase activity may correlate with the activity of lysosomal hydrolases and the concentration of sphingolipids. This study aimed to assess the association of the rs823144 variant in the promoter region of the RAB29 gene with PD risk, and to evaluate RAB29 expression, lysosomal hydrolase activity, and sphingolipid concentrations in the blood of PD patients. We screened the rs823144 variant of the RAB29 gene in a cohort of PD patients (N = 903) and controls (N = 618) using next-generation sequencing (NGS) and polymerase chain reaction (PCR) followed by restriction fragment length polymorphism analysis. The expression of the RAB29 gene was measured in peripheral blood mononuclear cells (PBMCs) using qPCR. We assessed the activities of lysosomal hydrolases (glucocerebrosidase (GCase), alpha-galactosidase (GLA), acid sphingomyelinase (ASMase), and galactosylcerebrosidase (GALC)) and the concentrations of sphingolipids (globotriaosylsphingosine (LysoGb3), sphingomyelin (LysoSM), and hexosylsphingosine (HexSph)) in blood using high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS). The RAB29 rs823144 C allele was associated with a reduced risk of PD in the Northwestern Russian population (OR = 0.7806, 95 % CI: 0.6578–0.9263, p = 0.0046), which is consistent with global data. However, no significant association was observed between the rs823144 C allele and RAB29 mRNA expression in PBMCs. Notably, the C allele was associated with increased GLA activity and decreased concentrations of LysoGb3 and LysoSM in the blood of PD patients. In conclusion, we demonstrate for the first time an association between the RAB29 rs823144 C allele and a reduced risk of PD in the Northwestern Russian population. Moreover, the RAB29 rs823144 C allele is associated with altered lysosomal enzyme activity and sphingolipid profiles, suggesting a potential role of RAB29 in sphingolipid metabolism relevant to PD pathogenesis.

Chronic obstructive pulmonary disease (COPD) is a multifactorial disease of the respiratory system and is the third leading cause of death worldwide. In the framework of the most relevant concepts of COPD pathogenesis, the key focus is on accelerated cellular senescence. FOXO family transcription factors are important hub components of cellular senescence signaling pathways. The objective of the study is to identify the association of FOXO1 (rs12585277, rs9549240), and FOXO3A (rs2253310, rs3800231) genes polymorphic variants with COPD and disease phenotypes. DNA samples from COPD patients (N = 710) and healthy individuals (N = 655) were used, polymorphic loci were analyzed by real-time PCR. For the first time, significant associations of FOXO1 (rs12585277) and FOXO3A (rs2253310) gene polymorphic loci with COPD and disease phenotypes were shown. Association with COPD was established with FOXO1 (rs12585277) (P_adj = 0.0018, OR = 1.44 for the AG genotype) and FOXO3A (rs2253310) (P_adj = 5.926 × 10^–7, OR = 1.99 for the GG genotype). A significant genotype-dependent variation of smoking index (in pack/years), vital capacity and forced vital capacity was revealed for FOXO1 (rs9549240, rs12585277) and FOXO3A (rs2253310) loci. Multiple regression and ROC analysis identified highly informative COPD risk model, which included polymorphic variants of the FOXO1 (rs12585277) and FOXO3A (rs2253310) genes, smoking index and age (P = 5.25 × 10^–93, AUC = 0.864). The multivariate regression model of the COPD “frequent exacerbator” phenotype included the AG genotype of FOXO1 (rs12585277), smoking index and age (AUC = 0.897, P = 4.1 × 10^–86). FOXO family transcription factors involved in autophagy, oxidative stress and cellular homeostasis may provide a platform for a new diagnostic and treatment strategy for COPD as potential biomarkers and targets for therapy.

To date, several genome-wide association studies (GWAS) of antisocial behavior (ASB) have been conducted in Europeans, which promoted research aimed at evaluating liability to ASB-related phenotypes in independent samples. Such studies implemented a polygenic score (PGS) approach, which represents a composite score considering a number of “risky” alleles. Since no GWAS of ASB has been conducted in Russians, the present study aimed to perform a replication study of liability to severe criminal behavior (homicide) in individuals from Russia using PGS. Moreover, we sought to obtain the best model considering PGS and potential social factors as predictors. Genotyping of the “top” ten SNPs previously identified in GWAS meta-analysis of ASB (CADM2, REV3L, FOXP1, FOXP2, BDNF, FURIN, XKR6, TMEM18, SORCS3, and ZIC4 genes) was conducted via real-time PCR in 227 homicide offenders and 254 healthy donors from the Volga-Ural region of Russia. Multiple regression models included “weighted” and “unweighted” PGS and potential social factors as predictors. The best regression model of liability to severe ASB was based on genetic effects of examined SNPs and social predictors, including traumatic brain injury, severe chronic disease, and tobacco smoking, which was more pronounced among subjects with a family history of mental illness (p = 2 × 10^–13). PGS alone explained a small proportion of variance in liability to ASB (1.1–1.5 %), while the inclusion of social parameters increased variance explained (16.2–21.2 %). Revealed findings evidence a higher impact of social factors than a composite effect of selected “top” SNPs in predicting liability to ASB in the examined cohort. A higher probability of ASB was linked to comorbid substance abuse, traumatic brain injury, and family history of mental illness, which may also represent a result of a “risky” genetic profile.

The neuropeptide oxytocin (OT) secreted by specialized neurons in the hypothalamus affects social behavior and aggression in various animal species in a dose-dependent manner. Our earlier studies showed that OT administration by nasal application to adult and adolescent Norway rat males selected for enhanced aggressive response to humans reduced aggression upon the opponent in the resident-intruder test. By contrast, OT administration to rats selected for tame behavior exerted no effect on behavior or even enhanced aggression. It was still unknown how selection for behavior affected the endogenous oxytocinergic system in rats. Here we study the populations of OT-containing cells in the paraventricular and supraoptic nuclei of the hypothalamus in intact tame and aggressive rats with regard to lateralization, as the hypothalamus is known to be functionally asymmetrical. We have also assessed blood OT changes after nasal OT application to rats selected for behavior. As it is known that the effect of OT on rat aggressiveness may depend on the basal level of the latter, we have analyzed the effect of OT administration on behavior in tame and aggressive rats interacting on neutral ground, where the aggressiveness of males manifests itself less than in the defense of territory in the resident-intruder test. The asymmetry in the numbers of OT-containing cells in the left and right halves of the paraventricular and supraoptic nuclei has been observed only in tame rats. The number of such cells in the right half of tame rats is greater than in aggressive. In contrast, the blood OT level in tame rats is significantly lower than in aggressive ones both in the intact animals and after OT administration. Oxytocin administration to aggressive rats shortens aggressive interactions and lateral threats and reduces the number of the latter as compared to animals of the same behavior pattern having received saline. This observation may point to an anti-aggressive effect of OT. In tame rats, though, OT administration increases the number of hind leg kicks and kicking duration. It appears that the differences in the endogenous OTergic system of hypothalamus found in this study are associated with both the behavior formed during selection and different responses to exogenous OT in tame and aggressive animals.

One of the main methods for obtaining transgenic plants is Agrobacterium-mediated transformation. This process relies on the ability of certain soil bacteria, specifically from the genera Agrobacterium and Rhizobium, to transfer and integrate a fragment of their plasmid into the chromosome of the recipient plant. This transferred DNA is referred to as T-DNA. Laboratory studies have demonstrated that whole plants can be regenerated from transgenic cells. It soon became evident that similar processes occur in nature, leading to the emergence of naturally transgenic plants, or natural GMOs. Thus, naturally transgenic plants possess homologues of the T-DNA genes from agrobacteria in their genomes (cellular T-DNA, or cT-DNA). These sequences are inherited through multiple sexual generations and retain their functionality. Furthermore, the potential for using newly acquired plant sequences in phylogenetic studies has been established, as cT-DNAs are clearly defined, highly specific, and recognizable DNA fragments that differ from typical plant DNA sequences. They are not found in untransformed ancestors, and their integration at specific chromosomal sites marks a monophyletic group of species. This review highlights the diversity of cellular T-DNAs and their potential use as phylogenetic markers. It includes a description of the main methodological approaches to such studies and discusses specific examples that clarify controversial points in the phylogeny of the genera Nicotiana, Camellia, Vaccinium, and Arachis. An important aspect of phylogenetic analysis based on cT-DNA is the assembly of individual alleles, which enables the tracking of interspecific hybridization events. This approach demonstrated the incomplete process of speciation within the Thea section of the genus Camellia and confirmed the role of interspecific hybridization in the breeding of North American blueberries. The review also addresses the dating of transformation events based on cT-DNA, which are organized in the form of imperfect repeats, as well as the application of phylogenetic studies to investigate the biodiversity of agrobacterial T-DNA genes.

In spite of recent substantial progress in genomic approaches, there is still a need for molecular markers convenient for Sanger sequencing and providing good phylogenetic reconstructions at short evolutionary distances. A new molecular marker, the histone H3–H4 region, containing partial coding sequences of the genes for histones H3 and H4 and the non-coding spacer between them, is proposed. This marker is potentially useful for molecular phylogenetic studies at the generic, species, and even intra-species level in insects and some other organisms, even from other phyla. The highly conserved histone-coding sequences ensure the universality of primers and the ease of primary alignment, while the highly variable non-coding spacer provides enough variation for analyses at short evolutionary distances. In insects, the histone genes reside in the histone repeat which is tandemly repeated in dozens to hundred copies forming the so-called histone cluster. This ensures a high concentration of the template for the marker in genomic DNA preparations. However, the order and orientation of the histone genes in the histone repeat is variable among orders, which puts some limitations on the use of the proposed marker. The marker efficacy is hereby shown for Odonata (dragonflies and damselflies), where it provided good resolution at the family, genus and species levels. The new marker also provided an interesting pattern in the relationship of two Sympetrum species, S. croceolum and S. uniforme, showing the sequences of the latter as a branch nested among those of the former. The same combination of the proposed original primers should also work in Diptera.

Lonicera L. is one of the largest and economically significant genera in the family Caprifoliaceae Juss., with a controversial taxonomy. To contribute to its molecular taxonomy, we sequenced the plastomes of Lonicera species: Lonicera caerulea (two subspecies), L. tatarica, and L. micrantha – using next-generation sequencing technology and conducted a comparative analysis. Plastome sizes ranged from 153,985 bp in L. micrantha to 164,000 bp in L. caerulea subsp. pallasii, each containing 130 genes, including 85 protein-coding, 37 tRNA, and 8 rRNA genes. Five protein-coding (rps7, rps12, ndhB, ycf2, and ycf15), 7 tRNA (trnA-UGC, trnI-CAU, trnI-GAU, trnL-CAA, trnN-GUU, trnR-ACG, and trnV-GAC), and 4 rRNA (rrn4.5, rrn5, rrn16, and rrn23) genes were duplicated. Comparative analysis of Lonicera plastome boundaries revealed structural variations in L. caerulea subsp. altaica and L. caerulea subsp. pallasii, particularly in ndhA gene distribution. Three highly variable, two intergenic (ycf1-trnN-GUU and trnN-GUU-ndhF) and one genic (accD) region were identified. A total of 641 simple sequence repeats were detected in four plastomes. Phylogenetic analyses grouped Lonicera samples into two clades corresponding to subgenera Periclymenum and Chamaecerasus. In this study, the plastid genomes of two subspecies of L. caerulea and species L. micrantha were sequenced for the first time. The maximum likelihood tree derived from complete plastid genome sequences proved to be the most informative, showing a topology consistent with previous studies. The nucleotide sequences of variable regions (accD-ycf1-ndhF-trnN-GUU) demonstrate high potential for use in DNA barcoding and may serve as valuable molecular markers for species phylogenetic studies within the genus Lonicera.

It typically takes 12 to 15 years to develop a new promising variety. One of the ways to reduce this time is through speed breeding. This method allows for up to six consecutive generations of spring cereals in a single year. Although far-red light is often overlooked in speed breeding protocols, it serves as a potent inducer of accelerated flowering in various plant species. In this study, we explored the advantages of far-red light as a means to optimize the speed breeding of spring triticale. Experimental plants were cultivated under three conditions with different red to far-red ratios at 660 nm (R – red) and 730 nm (FR – far red): 1) 3.75 (R > FR); 2) 0.8 (R = FR) and 3) 0.3 (R < FR). We found that the onset of triticale flowering occurred significantly earlier at the lowest red to far-red light ratio (R/FR 0.3). On average, plants bloomed 2.6 and 4.1 days earlier in a mineral wool and a soil mixture at R/FR 0.3, respectively, than those grown at R/FR 3.75. A negative effect of higher-intensity far-red light on the reproductive system of triticale was observed. Additionally, seeds obtained from plants grown under higher-intensity far-red light showed significantly lower germination energy and capacity. No differences were found in the regenerative capacity of isolated embryos in vitro obtained from plants grown under the different spectral compositions. Our results demonstrate that the accelerated triticale development requires not only the involvement of far-red light, but also a specific red to far-red light ratio close to 0.3. A modified speed breeding protocol relying on this ratio enabled flowering to commence as early as 33.9 ± 1.2 days after sowing. The same triticale variety grown under field conditions in the Krasnodar region and in traditional laboratory growing conditions with a photoperiod of 18/6 h day/night flowered 25 to 29 days later than those cultivated under the speed breeding conditions.