A variety of plant species found in nature contain agrobacterial T-DNAs in their genomes which they transmit in a series of sexual generations. Such T-DNAs are called cellular T-DNAs (cT-DNAs). cT-DNAs have been discovered in dozens of plant genera, and are suggested to be used in phylogenetic studies, since they are well-defined and unrelated to other plant sequences. Their integration into a particular chromosomal site indicates a founder event and a clear start of a new clade. cT-DNA inserts do not disseminate in the genome after insertion. They can be large and old enough to generate a range of variants, thereby allowing the construction of detailed trees. Unusual cT-DNAs (containing the rolB/C-like gene) were found in our previous study in the genome data of two Vaccinium L. species. Here, we present a deeper study of these sequences in Vaccinium L. Molecular-genetic and bioinformatics methods were applied for sequencing, assembly, and analysis of the rolB/C-like gene. The rolB/C-like gene was discovered in 26 new Vaccinium species and Agapetes serpens (Wight) Sleumer. Most samples were found to contain full-size genes. It allowed us to develop approaches for the phasing of cT-DNA alleles and reconstruct a Vaccinium phylogenetic relationship. Intra- and interspecific polymorphism found in cT-DNA makes it possible to use it for phylogenetic and phylogeographic studies of the Vaccinium genus.

In late December 2019, the first cases of COVID-19 emerged as an outbreak in Wuhan, China that later spread vastly around the world, evolving into a pandemic and one of the worst global health crises in modern history. The causative agent was identified as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Although several vaccines were authorized for emergency use, constantly emerging new viral mutants and limited treatment options for COVID-19 drastically highlighted the need for developing an efficient treatment for this disease. One of the most important viral components to target for this purpose is the main protease of the coronavirus (Mpro). This enzyme is an excellent target for a potential drug, as it is essential for viral replication and has no closely related homologues in humans, making its inhibitors unlikely to be toxic. Our review describes a variety of approaches that could be applied in search of potential inhibitors among plant-derived compounds, including virtual in silico screening (a data-driven approach), which could be structure-based or fragment-guided, the classical approach of high-throughput screening, and antiviral activity cell-based assays. We will focus on several classes of compounds reported to be potential inhibitors of Mpro, including phenols and polyphenols, alkaloids, and terpenoids.

The genus Vaccinium includes almost 500 species, among which there are economically important species of cranberries V. macrocarpon Ait. and V. oxycoccos L., lingonberries V. vitis-idaea L., bilberries V. myrtillus L. and blueberries V. uliginosum L., V. angustifolium Ait., V. corymbosum L., V. virgatum Ait. Despite the fact that many of these species were actively used by humans in medicine and food, their active selection began in the 20th century, in connection with which a classification of the genus according to morphological characters was developed. Many of these data remain relevant to the present day. The development of the ideas of molecular phylogeny prompted a revision of the old classification, identifying a number of difficulties that do not allow one to unambiguously determine phylogenetic relationships within the genus. Today, the genus includes 33 sections, while the species composition of the sections and the evolutionary relationships between them remain controversial. This review discusses various approaches to the study of the structure of the genus Vaccinium: from classical to phylogenomic, the main results of using these approaches and their prospects.

Genetic colonization by agrobacteria is possible due to agrobacterial transformation, which implies interspecies transfer of genetic material (T-DNA). A transgenic tissue is formed on the whole non-transgenic plant during that process. However, it turned out that in nature there are plants containing T-DNA fragments in their genomes and they can inherit these T-DNAs sexually. Such T-DNA was called cellular, and such plants were called natural transgenic. Examples of such organisms are plants of the genus Vaccinium. In the genomes of two species of this genus we found cT-DNA, represented by a rolB/C-like gene [1]. Previously, analyzing the natural transgenes in another genus (Camellia L.) [2], we showed the importance of reconstructing the allelic states of transgenes for phylogenetic studies. In this study, we performed analysis of the spreading of the rolB/C-like gene for its use as a molecular marker within Vaccinium. We used molecular-genetic and bioinformatics methods for sequencing, assembly, and analysis of the rolB/C-like gene. We discovered 26 new Vaccinium species and Agapetes serpens (Wight) Sleumer as containing the rolB/C-like gene. Most of studied samples are characterized by the presence of full-size genes. This made it possible to develop approaches for alleles phasing of the rolB/C-like gene and reconstruct a Vaccinium phylogenetic relationship. We subjected the studied species to phylogenetic analysis based on sequences of the rolB/C-like gene. The resulting phylogenetic tree of the genus Vaccinium divided the species into sections in accordance with the classical genus system based on morphological characters. At the same time, our tree did not confirm the taxa determined on the basis of ITS.

SARS-CoV-2 main protease (M^pro) is essential for viral polyprotein processing and represents a prime target for antiviral drug discovery. However, most available screening strategies rely on computational predictions or cell-free biochemical approaches that provide limited functional context and often require specialized instrumentation, while mammalian cell-based models remain costly and require high biosafety levels. Accordingly, there remains a shortage of simple, rapid, and biosafe functional screening tools suitable for early-stage prioritization of potential M^pro inhibitors, particularly those derived from natural sources and in urgent situations such as the COVID-19 pandemic. In this study, a bacterial colorimetric reporter assay was developed that directly links SARS-CoV-2 M^pro activity to β-galactosidase function in Escherichia coli. To the best of our knowledge, the developed assay represents the first bacterial colorimetric model for functional detection of SARS-CoV-2 M^pro inhibition based on a phenotypic readout. The assay enables the rapid visual detection of protease inhibition on X-gal-containing medium and provides a cost-effective and biosafe platform for prioritizing candidate inhibitors, under standard laboratory conditions, prior to further validation. Due to its bacterial expression context, this assay is intended for functional screening to provide the most promising candidate compounds and/or extracts for subsequent biochemical or mammalian cell-based validation; it is not intended to determine quantitative potency or to replace further validation approaches. It should be noted that the selective compound uptake in E. coli restricts the range of chemical compositions that can be evaluated using this method. Therefore, proof-of-concept application was demonstrated using pomegranate juice, a representative natural inhibitor source, rather than most currently known specific M^pro inhibitors. In addition, other plant-derived preparations, including rhubarb, grape, and red/black currant juices, were tested demonstrating the assay’s applicability to diverse natural matrices.

The two-spotted spider mite, Tetranychus urticae Koch, is a major agricultural pest with a rapid propensity for developing acaricide resistance. Bifenazate targets mitochondrial cytochrome b (CYTB). While the G126S mutation is associated with resistance, its independent role remains unclear, as it often occurs with other SNPs. This study explores the molecular basis of bifenazate resistance in a Russian laboratory strain derived from a St. Petersburg greenhouse population. Disruptive selection with increasing bifenazate concentrations generated resistant and susceptible isofemale lines. AlphaFold2 structural modeling of CYTB indicated that G126S causes a steric clash, leading to conformational destabilization, whereas other reported mutations primarily affect the ligand-binding pocket. Oxford Nanopore sequencing revealed a low initial frequency of the G126S allele (<1%; 226/35,895 reads) in the unselected population. After one year of stepwise selection (0.00005–0.031% a.i.), the mutant allele frequency surged to 90% (7272/8056 reads). No other resistance-associated mutations were found in the analyzed cytb fragment. We report the first identification of the G126S mutation in a Russian T. urticae population and demonstrate rapid fixation under bifenazate selection. Within this genetic background, G126S alone appears sufficient to confer high-level resistance, emphasizing the population-specific nature of resistance evolution and the critical need for local monitoring.