Phytophthora infestans is a highly destructive oomycete for Solanum tuberosum (potato), causing late blight disease with significant yield losses worldwide. While resistance (R) genes and targeted editing of susceptibility (S) genes have been widely investigated for disease management, the functional dynamics of S-genes during infection are less understood. In this study, we investigated cultivar-dependent differences in late blight susceptibility and the expression of S-genes during different infection stages. Desiree was the most susceptible cultivar, followed by Lady Rosetta, Sante, Cardinal, Asterix, Coroda, and Mozika. While no visual tuber blight symptoms were observed, infected plants produced a significantly higher number of smaller-sized tubers, which suggests the impact of foliage late blight. Among the candidate S-genes, expression analysis revealed that St-DMR6, St-PMR4, and St-CESA3 underwent consistent early induction (5 dpi) across all susceptible cultivars. However, by 15 dpi, expression patterns became cultivar-specific; induction was sustained in Cardinal and Desiree but suppressed in Coroda and Lady Rosetta. The early induction of S-gene(s) at 5 dpi likely depicts facilitated pathogen colonization, while sustained expression at 15 dpi indicated a stable role during later stages of infection. Our findings highlight the critical involvement of S‑genes, particularly St-PMR4, in mediating cultivar-specific susceptibility to P. infestans and provide potential targets for developing late blight-resistant potato through genome editing.
Plant viruses significantly decrease the yield of cultivated plants; therefore, developing new methods of combating viral diseases and studying the effectiveness of these methods is relevant. The expression of the heterologous extracellular RNase gene of Zinnia elegans (ZRNase II) increases the resistance of transgenic plants to viruses. The authors have proposed a model system for visualizing the protective effect of the ZRNase II gene and studying the spread of GFP-marked viral particles in Nicotiana benthamiana plants. The presence of the target ZRNase II gene was confirmed in the plants obtained after Agrobacterium-mediated transformation. Four lines were selected and adapted to ex vitro conditions. Total RNase activity in all transgenic lines was four to seven times higher than in wild-type plants. The systemic spread of the Potato virus X-based PVX-GFP construct in these plants was studied. To this end, agroinfiltration was performed using the pICH27566 and pICH6692 vectors. Following agroinfiltration, transient expression of GFP occurred and its systemic spread throughout the plant was observed due to transport proteins within the virus-like particles. Expression of the heterologous RNase gene (ZRNase II) delayed the spread of viral particles by 5–30 days and decreased the accumulation of GFP-marked viral particles in all transgenic lines compared to the control. To assess the rapid systemic response of plants to agroinfiltration and virus expression, the content of chlorophyll and anthocyanins in N. benthamiana leaves was studied outside the zone of visual spread of GFP-marked viral particles. Differences were found between control and transgenic plants in anthocyanin content in response to infection. Expression of the ZRNase II gene in plants delayed the systemic spread of virus-like particles and changed the response to biotic stress at the plant secondary metabolism level.
Growth regulating factor (GRF) binds to the QLQ (Gln, Leu, Gln) domain at the N-terminal of GRF interacting factor (GIF), acting as a transcriptional co-activator and playing important roles in plant growth, development and resistance to stresses. To date, no comprehensive identification and analysis of GRF and GIF gene families have been reported in Isatis indigotica Fort. In this study, the gene structures, conserved motifs, physicochemical properties, chromosomal localizations, cis-acting elements, evolutionary relationships and expression patterns were systematically analyzed. The results showed that there were 10 IiGRFs distributed on 5 chromosomes, and 4 IiGIFs on 2 chromosomes. IiGRFs and IiGIFs could be divided into 4 and 2 subfamilies, respectively. IiGRFs has two pairs of segmental duplication genes, but no tandem duplication genes. Moreover, IiGRFs (19.14-fold) and IiGIFs (8.0-fold) had the highest expression levels in young leaves and stems, compared with those in roots. The upstream regions of IiGRFs and IiGIFs contained more photo and MeJA hormone responsive elements, responding to shade and hormone stresses. Furthermore, the correlation analysis showed that IiGRF4 and IiGIF2, IiGRF4 and IiGIF4, and IiGRF4 and IiGIF3 were co-expressed with each other, which suggested that these paired genes could function together. The findings provided the more fundamental information on IiGRFs and IiGIFs, and laid good foundations for their further functional research in I. indigotica.
The results of the analysis of new genomic techniques (NGTs), currently widely used methods for genome editing (GE/GEd), are discussed. The CRISPR/Cas system, the most precise and efficient genome editing biotechnology, is considered in more detail. The rapid growth of plants improved by new editing methods has led to the need to consider or adopt new regulatory approaches to genome-edited plants. This review analyzes the regulatory landscape of genome-edited crops and products produced from them in some countries and regions of the world. A comparison is made with the legislation on genetically modified organisms. Current discussions and proposals for the legal regulation of genome-edited plants in the European Union are considered. Genome-editing methods and the regulatory framework for regulating genome-edited plants in the global world are constantly developing and changing, so the authors tried to appeal to the latest data, relying on academic publications and relevant regulatory documents. The publication aims to provide an overview of the various regulatory approaches currently in place (in use) or under consideration (under consideration) for genetically modified plants in some countries around the world.
Wool is a valuable and irreplaceable raw material for the textile industry, but a significant amount of it is defective. Improving the qualitative characteristics of wool is impossible without studying its structure. Accordingly, the purpose of this research was to study the structure of normal and felted wool of sheep of the Ukrainian Carpathian mountain breed. Samples of wool, which were divided into awn and down, were studied. The surface of the fibers was studied using scanning electron microscopy (SEM), while cross-sections were analyzed using transmission electron microscopy (TEM), and keratosis was examined through treatment with peracids and alkali. Studies have shown that the cortex of downy fibers is characterized by a bilateral structure, i.e., it consists of ortho- and paracortical cells, and the structure of the awn is characterized by radial asymmetry. A feature of the structure of the awn fibers is the presence of a medullary layer, which is located in the central part of the hair and is composed of porous tissue containing cavities. The cuticular layer of the awn, in comparison with the downy fibers, contains a larger number of scales that differ in their shape, and it is this layer that undergoes the greatest changes in felted wool, as indicated by the scales exfoliated in some places with smoothed edges and a deformed surface, as well as a significant decrease in the beta-keratosis fraction, both in the awn (from 15.28 to 11.98
Sodium-GLucose сoTransporter 2 (SGLT2) inhibitors, known as gliflozins, are a new effective class of hypoglycemic drugs. This study has revealed major variability in the pockets of the target site for all human SLC5 family members and confirmed the possibility of SGLT2 selective inhibition. Considering the uniqueness of the site, the virtual screening of Enamine Ltd chemical space identifies 36 prospective inhibitors of SGLT2. The ranking of selected compounds based on docking scor and binding energies identified 5 leaders with predicted activity comparable with known approved drugs. For the first time, the compounds Z2195993226 and Z2195993230 were identified as potential gliflozins. One more compound, Z1494829516 (Puerarin), previously known as an autophagy inducer, ferroptosis inhibitor, cardioprotector, antioxidant, anti-inflammatory, and antipyretic, was proposed as a potential gliflozin. Compound Z2417819595 was excluded from the list of possible SGLT2 inhibitors based on structural considerations. Furthermore, Z2235801995 was identified as the active compound of FDA-approved gliflozin NVOKAMET (FDA ID: 4129180), confirming the correctness of the screening protocol.
Osteoarthritis (OA) represents the most prevalent form of joint disease, leading to considerable pain and functional disability. While surgical interventions and pain relief modalities are available, there is an urgent necessity for effective therapeutic strategies to restore the damage. Our research investigates CK2.1, an innovative peptide that has exhibited significant promise in stimulating chondrogenesis, specifically within articular chondrocytes, for the treatment of OA. In our study, we employed micromass cultures derived from chondrocytes of human OA patients to evaluate the effectiveness of CK2.1. We demonstrated that CK2.1 significantly enhanced proteoglycan synthesis in chondrocytes isolated from OA patients. This observation highlights CK2.1’s targeted action and potential as a novel therapeutic strategy for treating osteoarthritis.
Thalassemia is the most prevalent form of inherited anemia throughout the world. It is estimated by the World Health Organization that approximately 60 000 babies are born with significant forms of thalassemia each year. This study uncovers the role of LRP5 (rs4988321, rs3736228) and VEGF (rs699947) genetic variants as genetic modifiers affecting vascular and skeletal complications by examining their frequency and association in β-thalassemia patients. The T allele of LRP5 gene SNP rs3736228 was more prevalent in patients (66
Artificial introgression hybridization within Triticeae is used to expand gene pools of cultivated species. Significant progress has been achieved, first of all, for traits of resistance to biotic stressors and product quality. The improvement of the methodical basis of molecular genetics now allows us to operate with the sequences of genome/transcriptome sequences and to compare hybrid genomes within the limits of their expression with the components of the original crosses. This changed the approach to evaluating the result of introgression. Traditionally, introgression was considered at the phenotypic level as a manifestation of the trait of interest and at the level of the nucleotide sequence of alien DNA present in the introgression line. In modern works, there is a gradual shift of attention from ascertaining the appearance of the target trait among the offspring to the study of the participation of epigenetic mechanisms in its implementation: DNA methylation and the participation of transposons and noncoding RNAs. These processes are activated by the very fact of combining two different genomes in one hybrid and originating an additional level of variability, which is reflected in the formation of the target phenotype. Currently, it is becoming obvious that changes at the level of nucleotide sequences and the organization of their expression can relate to both alien and genes of the recipient genome itself.
Prostate cancer (PCa) is considered the most common malignancy in men globally. Although prostate specific antigen (PSA) is still the only universally utilized biomarker related to prostate cancer, however PSA is not an ideal biomarker and its level can be altered by many factors. The aim of this study is to study serum H19 and miR-675 expression in prostate cancer patients and their potential diagnostic prognostic significance targeting to find a noninvasive biomarker. This study comprised 25 PCa patients and 25 healthy controls. The expression of serum H19 and miR-675 was detected by real-time polymerase chain reaction while N-Cadherin as an adhesion marker was measured by ELISA (enzyme-linked immunosorbent assay). The expression levels of serum H19 and miR-675 were increased in prostate cancer patients compared with controls. Also serum level of N-cadherin was higher in prostate cancer patients. Upregulation of H19 and miR-675 correlated with PCa metastasis and higher Gleason score. By using the ROC curve, the sensitivity of H19 and miR-675 was higher than PSA for the diagnosis of PCa. The results suggest that H19 and miR-675 expression could be considered potential noninvasive diagnostic and prognostic biomarkers for PCa.
Accipiter nisus is a small to medium-sized raptor widely distributed across Eurasia and parts of Africa. In this study, we sequenced the complete mitochondrial genome of A. nisus from Mianyang using the Illumina platform. We then compared mitochondrial genome A. nisus variations across four regions and analyzed the phylogenetic relationship of this species within the genus Accipiter. The results indicate that the mitochondrial genome of A. nisus from Mianyang is 18297 bp in length, which is smaller than those from Qufu (18647 bp), Yeongyang-gun (18352 bp), and Buk-Gu (19417 bp). The genomes of all four individuals contain 37 genes, an A+T-rich control region, and a pseudo-control region, with the most notable length variation occurring in the pseudo-control region. Additionally, inconsistencies in the usage of start and stop codons were observed in some protein-coding genes (PCGs). Phylogenetic analysis based on 13 PCGs and CYTB genes revealed that A. nisus is phylogenetically related to A. gentilis. This study provides essential genetic data and scientific evidence for A. nisus conservation.
Infertility mostly affects more than one member in a family, increasing the chances that the underlying cause is genetic. WGS aids in the discovery of novel variations through a variety of processes that cause infertility. To identify potential infertility-causing variations using WGS, a comprehensive method for analyzing the entire genome, and to identify the underlying mechanisms that may be targeted for future disease management. For this purpose, we identified a family with two male members exhibiting clinically diagnosed infertility and one fertile member. Blood samples were collected and subjected to WGS for CNV, SNV, and Run of homozygosity (ROH) analysis. The resulting WGS data were analyzed using bioinformatics tools/pipelines to investigate potential variants. We identified missense and stop-gain variations in TUBA3C and GJB2 genes as probable causes of Deafness Infertility Syndrome. Importantly, the TUBA3C variant represents the third deleterious variant linked to azoospermia in this emerging infertility gene. The TUBA3C and GJB2 variants were in a 9 and 18 kb region of overlapping ROHs in 12-1 and 12-2 (chr13:19138488-19231863) and (chr13:20060678-20434605), respectively. Furthermore, 12-3 did not have an ROH overlapping either variant. Families with two or more male individuals presenting with infertility, along with healthy fertile controls, present a vital resource and opportunity to comprehensively identify the genetic landscape in the maintenance of male fertility. This study will help in the mechanistic understanding of Deafness Infertility Syndrome and provide for identifying with certainty causal variations among idiopathic patients, targets for management, and the development of future therapy for male infertility.
Galinsoga parviflora and G. quadriradiata are among the most successful invasive species of the family Asteraceae, having actively spread across Europe and other continents since the late 18th century. Despite the significant impact on agroecosystems, the genetic structure of their populations remains insufficiently studied using molecular approaches. Using sequencing and bioinformatic analysis, we characterized the polymorphism of the chloroplast DNA intergenic spacer rpl32–trnL(UAG) in Galinsoga samples from Ukraine, other European countries, and China. It was found that the vast majority of G. parviflora and G. quadriradiata samples are represented by two main haplotypes, Par1 and Qua1, indicating low genetic diversity of these species within their secondary range and supporting the hypothesis of a single introduction involving a limited amount of source material. Four rare haplotypes were also identified, which likely originated within the invaded range. It was shown that in hybridization between G. parviflora and G. quadriradiata both species can act as the maternal parent. The effectiveness of the rpl32–trnL(UAG) region for genetic barcoding was confirmed, and it was found that G. parviflora dominate in the central and northern regions of Ukraine, while G. quadriradiata prevails in the western regions of the country.
The olfactory analyzer is a relatively simple, but important part of the nervous system for realization of many mental functions, the studies of which can significantly deepen knowledge about molecular mechanisms of chemical reception, regulation of gene expression, formation of new neurons, and development of neural networks in the mature brain, as well as about the pathogenesis of neurodegenerative and mental disorders. The article considers the structure and functions of the olfactory tubercle, diagonal band of Broca, and nucleus basalis of Meynert located within the so-called olfactory cortex. Based on numerous literary data, it was demonstrated that despite a relatively simple neural structure, all three these structures are involved in the mechanisms of the most complex mental functions, and the zone of their location is a kind of anatomical bridge between the anterior cingulate and anterior insular cortex, the significance of which in the mental process is difficult to overestimate. Along with the consideration of these issues, the article reveals a historical priority of Volodymyr Betz in the cytoarchitectonic description of the olfactory tubercle. It was also demonstrated that the work of V. Betz can contain information about the structure of the diagonal band and nucleus basalis, previously discovered by P. Broca and T. Meynert, respectively. Consequently, taking into account a significance of the considered part of the brain, cytoarchitectonic data V. Betz on the structure of the olfactory tubercle and adjacent structures can be considered no less important than his other observations recognized by a professional community.
In this study, a comparative analysis of the amino acid sequences and domain organization of histone deacetylases (HDACs) from Arabidopsis thaliana and Oryza sativa was performed, and the presence of experimentally determined 3D structures as well as the quality of predicted models was assessed. The results of the analysis of deacetylase inhibitor binding across different classes demonstrated their limited selectivity toward plant HDACs. Notable inhibitory selectivity was observed for SAHA, whereas trichostatin A, sodium butyrate, and cyclic hydroxamic acid derivatives acted predominantly as pan-HDACis (HDAC inhibitors). It was shown that sodium butyrate can bind within the catalytic pocket of deacetylases; however, its interaction is unstable and likely induces inhibition by triggering conformational changes. The structural analysis provided a detailed characterization of the shared and distinct features of the HDACs examined. HDAC10 from O. sativa was found to exhibit the highest structural similarity to HDA14 from A. thaliana, suggesting a potentially similar functional role and shared deacetylation substrates. Four key amino acid residues required for Zn2+ coordination in the active site were identified, including the “XDXH” motif, which is critically important for catalytic activity. The results obtained enhance our understanding of the structural organization of plant HDACs, identify key determinants of their catalytic activity, and provide a foundation for the development of selective inhibitors and biotechnological strategies aimed at regulating plant growth and improving stress tolerance.
The study was aimed at assessing the efficiency of SSR markers for the identification and differentiation of Ukrainian hazelnut varieties. The studied material included 30 specimens of Corylus spp., comprising 25 hazelnut variaties and three Corylus species from the collection of the Sofiyivka National Dendrological Park (National Academy of Sciences of Ukraine). The polymorphism and informativeness parameters of nine microsatellite (SSR) loci were analyzed. All loci were found to be polymorphic, with the number of alleles per locus ranging from four to seven. The informativeness indices were as follows: polymorphic information content (PIC) ranged from 0.337 to 0.760 (mean 0.617); expected heterozygosity (He) ranged from 0.367 to 0.791; observed heterozygosity (Ho) ranged from 0.240 to 0.920. The results indicate the high efficiency of the analyzed SSR loci as molecular markers for the identification and differentiation of Ukrainian hazelnut variaties and their potential use in breeding programs.
During this study, two consanguineous Pakistani families with symptoms of intellectual disability and primary microcephaly were investigated. Affected individuals were born to phenotypically normal parents. Clinical information, including age, sex, age at walking, short stature, microcephaly, behavioral severity, intellectual disability, facial dysmorphism, muscular hypertonia, and the existence of any other illnesses, such as speech delay and epilepsy, was documented. To identify disease-causing variants, whole exome sequencing and subsequent Sanger sequencing were performed. As a result, in one of the families, a novel homozygous missense substitution in the NSUN2 gene and a stop codon in the ASPM gene in the other family were identified. Sequence analysis confirmed a homozygous pathogenic variant (c.1853G>T, p.Arg618Ile) in NSUN2 (NM_017755.6) and a stop codon (c.3978G>A, p.Tryp1326X) in ASPM (NM_018136.5). Our results broadened the mutational spectrum of both genes (NSUN2 and ASPM) causing neurological disorders in the affected individuals of Pakistani families.
Recently, progress has been made in establishing the molecular mechanisms of inducing plant resistance to pathogens by β-aminobutyric acid (BABA), which is considered one of the new plant stress metabolites. However, the mechanisms and even the phenomenology of BABA’s effect on plant resistance to major abiotic stressors remain insufficiently studied. This study aimed to determine the effect of priming wheat grains (Triticum aestivum L.) with BABA on their germination under osmotic (12
Low-molecular-weight heat shock proteins (sHSPs) play a key role in protecting plants from abiotic stress and are also involved in growth and development processes. In this study, using bioinformatics approaches, a comprehensive analysis of the multigenic sHsp family in members of the Solanaceae family, which includes numerous food, medicinal, and ornamental plant species, was performed. As a result of a genomic screening for two members of this family, Nicotianaattenuata (coyote tobacco), Solanumlycopersicum (tomato), and the model plant Arabidopsisthaliana, 52, 45, and 34 putative sHsp genes were identified, respectively, and their chromosomal localization was determined. All tobacco genes, five tomato genes, and one Arabidopsis gene were identified for the first time. The putative sHSPs belong to 19 classes. They differ in the size of their structural domains and contain targeting signals for various cellular compartments. Most classes formed before the divergence of the phylogenetic lineages of Superrosids and Superasterids and were conserved throughout the further evolution of dicots, while some classes proved to be specific to A.thaliana or members of the Solanaceae family. Several sHsps are characterized by a clustered organization on the chromosomes, which arose through tandem duplications of individual genes during evolution. In the genomes examined, 15 sHsps were identified, encoding two to four alternative transcripts. Analysis of the promoter regions revealed numerous recognition sites for transcription factors associated with the regulation of stress responses, light signaling, phytohormone action, and various processes governing plant growth and development.
Low-molecular-weight heat shock proteins (sHSPs) play a key role in protecting plants from abiotic stress and are also involved in growth and development processes. In this study, using bioinformatics approaches, a comprehensive analysis of the multigenic sHsp family in members of the Solanaceae family, which includes numerous food, medicinal, and ornamental plant species, was performed. As a result of a genomic screening for two members of this family, Nicotiana attenuata (coyote tobacco), Solanum lycopersicum (tomato), and the model plant Arabidopsis thaliana, 52, 45, and 34 putative sHsp genes were identified, respectively, and their chromosomal localization was determined. All tobacco genes, five tomato genes, and one Arabidopsis gene were identified for the first time. The putative sHSPs belong to 19 classes. They differ in the size of their structural domains and contain targeting signals for various cellular compartments. Most classes formed before the divergence of the phylogenetic lineages of Superrosids and Superasterids and were conserved throughout the further evolution of dicots, while some classes proved to be specific to A. thaliana or members of the Solanaceae family. Several sHsps are characterized by a clustered organization on the chromosomes, which arose through tandem duplications of individual genes during evolution. In the genomes examined, 15 sHsps were identified, encoding two to four alternative transcripts. Analysis of the promoter regions revealed numerous recognition sites for transcription factors associated with the regulation of stress responses, light signaling, phytohormone action, and various processes governing plant growth and development.
