Employment of pqqE gene as molecular marker for the traceability of Gram negative phosphate solubilizing bacteria associated to plants.

Insoluble phosphorous compounds solubilization by soil bacteria is of great relevance since it puts available the phosphorus to be used by plants. The production of organic acids is the main microbiological mechanism by which insoluble inorganic phosphorus compounds are solubilized. In Gram negative bacteria, gluconic acid is synthesized by the activity of the holoenzyme glucose dehydrogenase-pyrroloquinoline quinine named GDH-PQQ. The use of marker genes is a very useful tool to evaluate the persistence of the introduced bacteria and allow to follow-up the effect of biotic and abiotic factors on these beneficial microorganisms in the soil. In previous studies we detected the presence of the pqqE gene in a great percentage of both non-culturable and culturable native soil bacteria. The objective of this study was to analyze the phylogeny of the sequence of pqqE gene and its potential for the study of phosphate solubilizing bacteria from pure and mixed bacterial cultures and rhizospheric soil samples. For this, the presence of the pqqE gene in the genome of phosphate solubilizing bacteria that belong to several bacteria was determined by PCR. Also, this gene was analyzed from mixed bacterial cultures and rhizospheric soil associated to peanut plants inoculated or not with phosphate solubilizing bacteria. For this, degenerate primers designed from several bacterial genera and specific primers for the genus Pseudomonas spp., designed in this study, were used. DNA template used from simple or mixed bacterial cultures and from rhizospheric soil samples was obtained using two different DNA extraction techniques. Results indicated that pqqE gene amplification product was found in the genome of all Gram negative phosphate solubilizing bacteria analyzed. It was possible to detect this gene in the DNA obtained from mixed cultures where these bacteria grew in interaction with other microorganisms and in that obtained from rhizospheric soil samples inoculated or not with these bacteria. The phylogenetic analysis indicated that pqqE gene is a conserved gene within related genera. In conclusion, pqqE gene could be a potential marker for the study of phosphate solubilizing bacterial populations.

The heart microbiome of insectivorous bats from Central and South Eastern Europe.

Host associated microbiome not only may affect the individual health-status or provide insights into the species- or group specific bacterial communities but may act as early warning signs in the assessment of zoonotic reservoirs, offering clues to predict, prevent and control possible episodes of emerging zoonoses. Bats may be carriers and reservoirs of multiple pathogens such as viruses, bacteria and parasites, showing in the same time robust immunity against many of them. The microbiota plays a fundamental role on the induction, training and function of the host immune system and the immune system has largely evolved in order to maintain the symbiotic relationship of the host with these diverse microbes. Thus, expanding our knowledge on bat-associated microbiome it can be usefully in understanding bats' outstanding immune capacities. The aim of this study was to investigate the presence of different bacterial communities in heart tissue of insectivorous bats, Nyctalus noctula, Pipistrellus pipistrellus and Rhinoplophus hipposideros, from Central and Eastern Europe using high-throughput sequencing of variable regions of the 16S rRNA. In addition, species-specific PCRs were used to validate the presence of the vector-borne pathogens Bartonella spp. and Rickettsia spp. In this study we identified a wide variety of bacterial groups, with the most abundant phyla being Proteobacteria and Firmicutes. The results showed that at individual level, the year or location had no effect on the diversity and composition of the microbiome, however host species determined both structure and abundance of the bacterial community. We report the presence of vector-borne bacteria Bartonella spp. in samples of N. noctula and indications of Rickettsia spp. in R. hipposideros. Our results provide a first insight into the bacterial community found in heart tissue of bats from Central and South Eastern Europe.

Evaluation of native bacteria and manganese phosphite for alternative control of charcoal root rot of soybean.

Plant growth promoting rhizobacteria (PGPR) are potential agents to control plant pathogens and their combined use with biopesticides such as phosphites may constitute a novel strategy to incorporate in disease management programs. In the present study, 11 bacterial isolates were selected on the basis of their antagonistic activity against Macrophomina phaseolina in dual-culture tests, and their plant growth promoting traits. Selected isolates were characterised on the basis of auxin and siderophore production, phosphate solubilisation and rep-PCR genomic fingerprinting. Two of these isolates, identified as Pseudomonas fluorescens 9 and Bacillus subtilis 54, were further evaluated for their inhibitory capacity against M. phaseolina using in vitro (on soybean seeds) and in vivo (greenhouse assay) tests. Both bacteria were applied individually as well as in combined treatment with manganese phosphite as seed treatments. Damage severity on soybean seeds was significantly reduced, compared with the untreated control, by both bacterial strains; however, the individual application of phosphite showed to be least effective in controlling M. phaseolina. Interestingly, the phosphite treatment improved its performance under greenhouse conditions compared to the results from the in vitro assays. In the greenhouse trials, the greatest reductions in disease severity were achieved when strain P. fluorescens 9 was applied singly or when strain B. subtilis 54 was combined with manganese phosphite, achieving 82% of control in both cases. This work is the first to report the control of M. phaseolina using combined treatment with PGPR and phosphite under greenhouse conditions.