Phylogenetic and pangenomic analyses of members of the family Micrococcaceae related to a plant-growth-promoting rhizobacterium isolated from the rhizosphere of potato (Solanum tuberosum L.).

We report the results of taxonomic studies on members of the family Micrococcaceae that, according to the 16S rRNA, internal transcribed spacer 1 (ITS1), average nucleotide identity (ANI), and average amino acid identity (AAI) tests, are related to Kocuria rosea strain RCAM04488, a plant-growth-promoting rhizobacterium (PGPR) isolated from the rhizosphere of potato (Solanum tuberosum L.). In these studies, we used whole-genome phylogenetic tests and pangenomic analysis. According to the ANI > 95 % criterion, several known members of K. salina, K. polaris, and K. rosea (including K. rosea type strain ATCC 186T) that are related most closely to isolate RCAM04488 in the ITS1 test should be assigned to the same species with appropriate strain verification. However, these strains were isolated from strongly contrasting ecological and geographical habitats, which could not but affect their genotypes and phenotypes and which should be taken into account in evaluation of their systematic position. This contradiction was resolved by a pangenomic analysis, which showed that the strains differed strongly in the number of accessory and strain-specific genes determining their individuality and possibly their potential for adaptation to different ecological niches. Similar results were obtained in a full-scale AAI test against the UniProt database (about 250 million records), by using the AAI-profiler program and the proteome of K. rosea strain ATCC 186T as a query. According to the AAI > 65 % criterion, members of the genus Arthrobacter and several other genera belonging to the class Actinomycetes, with a very wide geographical and ecological range of sources of isolation, should be placed into the same genus as Kocuria. Within the paradigm with vertically inherited phylogenetic markers, this could be regarded as a signal for their following taxonomic reclassification. An important factor in this case may be the detailing of the gene composition of the strains and the taxonomic ratios resulting from analysis of the pangenomes of the corresponding clades.

Improving the efficacy of potato clonal micropropagation by inoculation with the rhizosphere bacteria Azospirillum baldaniorum Sp245 and Ochrobactrum cytisi IPA7.2.

Sustainable development of agriculture depends on the provision of quality seeds to the market. Inoculation with plant-growth-promoting rhizobacteria in in vitro culture can be used to improve the growth eff icacy and performance of microplants. We examined the effect of in vitro inoculation of microplants of the cultivars Nevsky and Kondor with the strains Azospirillum baldaniorum Sp245 and Ochrobactrum cytisi IPA7.2 separately and in combination. We examined the morphological variables of plant growth in in vitro culture and under ex vitro adaptation conditions; we also investigated the growth and performance of the plants in the greenhouse. The dependence of the inoculation eff icacy on potato genotype, growth stage, and inoculum composition was ascertained throughout the experiment. In vitro, A. baldaniorum Sp245 alone and in combination with O. cytisi IPA7.2 promoted the formation of roots on the microplants of both cultivars and the growth of Nevsky shoots. During plant growth ex vitro, all growth variables of the Nevsky microplants were promoted by O. cytisi IPA7.2 alone and in combination with A. baldaniorum Sp245. In both cultivars grown in the greenhouse, shoot growth was promoted in most inoculation treatments. The survival ability of the Nevsky microplants in the greenhouse increased 1.7-fold under the effect of simultaneous inoculation. Inoculation of microplants with a combination of A. baldaniorum Sp245 and O. cytisi IPA7.2 increased the number of Nevsky minitubers 1.5-fold and the number of Kondor minitubers 3.5-fold. Inoculation with the tested strains can be used to promote the growth of microplants and increase the yield of minitubers in potato seed breeding for the production of healthy planting material.

Preparation of polyclonal antibodies to diminazene and its detection in animal blood plasma.

This work was undertaken to examine the possibility of preparing polyclonal antibodies to diminazene by using colloidal gold nanoparticles as an antigen carrier and adjuvant. The antibodies prepared by us were used for immunodot determination of diminazene in calf-blood plasma. The immunochemical results agreed with those obtained by high-performance liquid chromatographic estimation of diminazene content in animal biological fluids.

Obtainment of polyclonal antibodies to clenbuterol with the use of colloidal gold.

The effectiveness of an in vivo synthesis of antibodies to clenbuterol with the use of gold nanoparticles as a carrier was evaluated. For comparison, conjugates of clenbuterol with bovine serum albumin were used in immunization. The serum titer was determined by an ELISA. The antibodies were tested by an immunodot assay with immunogold markers. With both techniques we obtained specific and relatively high-titer antibodies to clenbuterol. It was found that the antibodies obtained with the use of gold nanoparticles were not inferior in titer to those obtained to the conjugates of clenbuterol with the protein but surpassed them in specificity.

The effectivity analysis of accumulation of liposomal, micellar, and water-soluble forms of diminazene in cells and in organs.

The study was aimed at evaluating the effectiveness of liposomal, micellar, and water-soluble drug forms of diminazene for its localization in cells and selective accumulation at the sites of aggregation of pathogenic organisms. Pharmacological and dynamic properties of a new injection micellar diminazene preparation were experimentally determined. These properties were compared with the same parameters obtained for the water-soluble and liposomal diminazene aceturate drug forms. The drug forms studied may be arranged in the following order of decreasing effectiveness of accumulation of diminazene in red and white blood cells and in murine organs: liposome form, micellar form, and water-soluble form.