In nature, plants are constantly exposed to a varied abiotic and biotic stresses or their combinations, limiting the productivity of major crops, including wheat. Combinations of drought and soil-borne Fusarium-instigated diseases are the most common combinations of stresses, significantly reducing wheat yield around the world. Here, were analyzed the potential of application of endophytic bacteria Bacillus subtilis (strain 10–4) together with the natural signal molecule salicylic acid (SA) to improve growth and tolerance of Triticum aestivum L. (wheat) plants under combined drought and Fusarium culmorum-instigated root rot (FRR) stresses. It was revealed that pre-sowing treatment with B. subtilis 10–4, SA, and B. subtilis 10–4 + SA, both under normal and combined drought conditions, notably reduced (by 50–80% or more) the incidence of FRR development in wheat plants, with the most notable effect for B. subtilis 10–4 + SA (wherein disease symptoms were almost absent). Moreover, B. subtilis 10–4, SA, and especially B. subtilis 10–4 + SA increased plant growth (root and shoot length, fresh and dry biomass) under normal (up to 20–50%), drought (up to 15–40%), FRR (up to 15–30%), and combined drought + FRR stresses (up to 20%), with the maximum effect for B. subtilis 10–4 + SA. Additionally, B. subtilis 10–4, SA, and B. subtilis 10–4 + SA decreased stress (drought, FRR, and combined drought + FRR)-instigated lipid peroxidation and osmotic damages of plant cells. The findings indicate that endophytic bacteria B. subtilis 10–4 alone and in a mixture with SA may be used as an effective eco-friendly agent to improve wheat growth and tolerance under the influence of drought, FRR, and combinations of these stresses.

Biomimetic patterning emerges as a promising antibiotic-free approach to protect medical devices from bacterial adhesion and biofilm formation. The main advantage of this approach lies in its simplicity and scalability for industrial applications. In this study, we employ it to produce antibacterial coatings based on silicone materials, widely used in the healthcare industry. In doing so, we patterned silicone substrates with a topography of various flower petals (rose, chamomile, pansy, and magnolia) and studied the relationship between the antibacterial properties of the obtained biomimetic substrates and their surface topography. To study the surface topography of biomimetic surfaces, we used the fractal analysis of their SEM images. In particular, as a measure of surface complexity and heterogeneity, we used the values of the developed interfacial area ratio (Sdr) and lacunarity coefficient (β). In the result, we found that the bacterial area coverage of biomimetic substrates decreased exponentially with the increase in their surface complexity and heterogeneity, and prominent antibacterial properties were observed at β > 1.6 and Sdr > 50. The results of this study can be used to identify biomimetic materials with superior antibacterial properties and produce efficient antibacterial silicone coatings for biomedical and healthcare applications.

Background. Rhizobia are the most effective nitrogen-fixing organisms that can fix nitrogen only in symbiosis with leguminous plants. The general transcriptional activator of nitrogen fixation genes in diazotrophic bacteria is NifA. In this work, the possibility of modifying the regulation of nitrogen fixation in the nodule bacteria Mesorhizobium, Ensifer and Rhizobium was studied by introducing an additional copy of the nifA gene into the bacterial genomes during the regulation of induced bacterial promoters. Materials and methods. A series of expression genetic constructs with NifA genes of nodule bacteria strains under the control of an inducible promoter Pm were created. The resulting constructs were transformed into strains of nodule bacteria. The obtained recombinant strains were investigated for the appearance of their nitrogen-fixing activity in the free-living state. Results. It was shown that the expression of nifA in recombinant cells of all three genera of bacteria leads to the appearance of insignificant nitrogenase activity. At the same time, the level of nitrogenase activity does not have a correlation with the level of expression of the introduced nifA gene, which, most likely, is a consequence of the multilevel regulation of nitrogen fixation. Conclusion. The possibility of artificial activation of nitrogenase activity in nodule bacteria in the free-living state by introducing the NifA regulatory protein gene into bacteria was shown.

The functional activity of different phylogenetic variants of the transcriptional activator of nitrogen fixation genes in the cells of root nodule bacteria (rhizobia) of different taxa was analyzed. For this purpose, recombinant strains of root nodule bacteria of the genera Mesorhizobium, Ensifer (Sinorhizobium), and Rhizobium were created using genetic engineering constructs based on the plasmid pJN105. This plasmid contains different phylogenetic variants of the nifA gene typical for bacteria of the genera Mesorhizobium, Ensifer, and Rhizobium under the control of the ParaBAD bacterial promoter. It was shown that in free-living recombinant cells of all three bacterial genera, the introduction of an additional active copy of the nifA gene to their genome resulted in emergence of detectable nitrogenase activity. The level of nitrogenase activity was not directly related to the phylogeny of the introduced nifA gene, which is most likely explained by the universality of the product of this gene within the studied group of bacteria. The data obtained indicate the possibility of combinatorial processes in the evolution of nitrogen fixation genes.

BACKGROUND: The beginning of the life cycle of a leguminous plant in its natural habitat is usually associated with interaction with nodule bacteria in order to form a nitrogen-fixing symbiosis. In a short period of time, a plant needs to “choose” suitable microsymbionts for itself. Since a wide variety of rhizobial strains is formed in the rhizosphere of legumes, the choice made by the macrosymbiont will further influence its productivity. AIM: The purpose of our work was to compare the principles of selection by different plants of their microsymbionts at different stages of plant development. MATERIALS AND METHODS: Nodule bacteria Trifolium hybridum L. and Galegaorientalis Lam. were taken into the study. Their genetic diversity was studied by the RAPD method, a phylogenetic analysis of bacteria and their symbiotic nodC and nifH genes was carried out, and their nitrogen-fixing activity was assessed. RESULTS: It was found that the rhizobia that form nodules on the roots of the studied leguminous plants at different stages of their vegetation have certain patterns. It was found that the highest polymorphism and specific nitrogen-fixing activity are characteristic of bacteria obtained from nodules formed at the initial stage of vegetation. CONCLUSIONS: We assume that the plasticity of the rhizobia genome gives the host plant the ability to more flexibly adjust its nitrogen-fixing apparatus to changes in growing conditions.

Background. An analysis of the spatial distribution of some taxonomically and ecologically related legumes in the Ural showed a nontrivial spatial distribution of related species of the genus Oxytropis DC of the Orobia Bunge section within the Uchalinsky uplands. Despite the similarities in ecology, these species practically do not grow together. Explicit spatial segregation of closely related plants over a relatively small area allows this phenomenon to be used as a convenient model for studying the effect of segregation of closely related legume species on the genetic composition of their nodule bacteria. Materials and methods. The genetic diversity of nodule bacteria entering into symbiosis with O. kungurensis, O. baschkiriensis, O. approximata and O. gmelinii plants was studied. In addition, the polymorphism of their symbiotic genes has also been analyzed. Results. Phylogenetic characteristics of nodule bacteria endemic for the Southern Ural belonging to 4 species of leguminous plants of the genus Oxytropis of the section Orobia: O. kungurensis, O. baschkiriensis, O. approximata, O. gmelinii which are characterized by spatial separation of the growth sites, also called plant segregation, are given. It was shown that all of them belong to the genus Mesorhizobium despite certain phylogenetic differences of bacteria. Analysis of the symbiotic genes of the analyzed strains revealed a lack of congruence of their phylogeny with the core part of the genome. It was found that the microsymbionts of O. baschkiriensis plants differ in the phylogeny of nod-genes from nodule bacteria of other plants of the Oxytropis genus and are close to microsymbionts of plants of the Lupinaster genus growing in the Southern Urals. Conclusion. Acquisition of the property to enter into symbiosis with nodule bacteria of plants of the genus Lupinaster may turn out to be an adaptive mechanism that arose as a result of segregation of O. baschkiriensis from other species of Oxytropis.