Bacteria Associated With a Commercial Mycorrhizal Inoculum: Community Composition and Multifunctional Activity as Assessed by Illumina Sequencing and Culture-Dependent Tools.

The implementation of sustainable agriculture encompasses practices enhancing the activity of beneficial soil microorganisms, able to modulate biogeochemical soil cycles and to affect soil fertility. Among them, arbuscular mycorrhizal fungi (AMF) establish symbioses with the roots of most food crops and play a key role in nutrient uptake and plant protection from biotic and abiotic stresses. Such beneficial services, encompassing improved crop performances, and soil resources availability, are the outcome of the synergistic action of AMF and the vast communities of mycorrhizospheric bacteria living strictly associated with their mycelium and spores, most of which showing plant growth promoting (PGP) activities, such as the ability to solubilize phosphate and produce siderophores and indole acetic acid (IAA). One of the strategies devised to exploit AMF benefits is represented by the inoculation of selected isolates, either as single species or in a mixture. Here, for the first time, the microbiota associated with a commercial AMF inoculum was identified and characterized, using a polyphasic approach, i.e., a combination of culture-dependent analyses and metagenomic sequencing. Overall, 276 bacterial genera were identified by Illumina high-throughput sequencing, belonging to 165 families, 107 orders, and 23 phyla, mostly represented by Proteobacteria and Bacteroidetes. The commercial inoculum harbored a rich culturable heterotrophic bacterial community, whose populations ranged from 2.5 to 6.1 × 106 CFU/mL. The isolation of functional groups allowed the selection of 36 bacterial strains showing PGP activities. Among them, 14 strains showed strong IAA and/or siderophores production and were affiliated with Actinomycetales (Microbacterium trichotecenolyticum, Streptomyces deccanensis/scabiei), Bacillales (Bacillus litoralis, Bacillus megaterium), Enterobacteriales (Enterobacter), Rhizobiales (Rhizobium radiobacter). This work demonstrates for the first time that an AMF inoculum, obtained following industrial production processes, is home of a large and diverse community of bacteria with important functional PGP traits, possibly acting in synergy with AMF and providing additional services and benefits. Such bacteria, available in pure culture, could be utilized, individually and/or in multispecies consortia with AMF, as biofertilizers and bioenhancers in sustainable agroecosystems, aimed at minimizing the use of chemical fertilizers and pesticides, promoting primary production, and maintaining soil health and fertility.

The Role of Polyphenoloxidase, Peroxidase, and ß-Glucosidase in Phenolics Accumulation in Olea europaea L. Fruits under Different Water Regimes.

Olive fruits and oils contain an array of compounds that contribute to their sensory and nutritional properties. Phenolic compounds in virgin oil and olive-derived products have been proven to be highly beneficial for human health, eliciting increasing attention from the food industry and consumers. Although phenolic compounds in olive fruit and oil have been extensively investigated, allowing the identification of the main classes of metabolites and their accumulation patterns, knowledge of the molecular and biochemical mechanisms regulating phenolic metabolism remains scarce. We focused on the role of polyphenoloxidase (PPO), peroxidase (PRX) and ß-glucosidase (ß-GLU) gene families and their enzyme activities in the accumulation of phenolic compounds during olive fruit development (35-146 days after full bloom), under either full irrigation (FI) or rain-fed (RF) conditions. The irrigation regime affected yield, maturation index, mesocarp oil content, fruit size, and pulp-to-pit ratio. Accumulation of fruit phenolics was higher in RF drupes than in FI ones. Members of each gene family were developmentally regulated, affected by water regime, and their transcript levels were correlated with the respective enzyme activities. During the early phase of drupe growth (35-43 days after full bloom), phenolic composition appeared to be linked to ß-GLU and PRX activities, probably through their effects on oleuropein catabolism. Interestingly, a higher ß-GLU activity was measured in immature RF drupes, as well as a higher content of the oleuropein derivate 3,4-DHPEA-EDA and verbascoside. Activity of PPO enzymes was slightly affected by the water status of trees during ripening (from 120 days after full bloom), but was not correlated with phenolics content. Overall, the main changes in phenolics content appeared soon after the supply of irrigation water and remained thereafter almost unchanged until maturity, despite fruit growth and the progressive decrease in pre-dawn leaf water potential. We suggest that enzymes involved in phenolic catabolism in the olive fruit have a differential sensitivity to soil water availability depending on fruit developmental stage.

Vanillin production by recombinant strains of Escherichia coli

A produção de vanilina a partir de ácido ferúlico foi estudada utilizando-se diferentes linhagens recombinantes de Escherichia coli. Para prevenir a ocorrência de condições de aerobiose e a possível oxidação do produto, os ensaios foram realizados em frascos Erlenmeyer sob agitação moderada (150 rpm). E. coli JM109 (pBBI) mostrou-se o melhor produtor de vanilina entre os demais agentes transformantes, sendo capaz de converter 95% do ácido ferúlico inicial em produto após 1h, rendimento este que decresceu para 72% após 72h, provavelmente devido à atividade de uma oxidase não-específica responsável pela oxidação de vanilina a ácido vanílico.

Ribosomal DNA sequence analysis shows that the basidiomycete C30 belongs to the genus Trametes.

The basidiomycete C30 was considered as an isolate of a population of Marasmius quercophilus collected on evergreen oak litter from the Mediterranean forest. Recent phenotypic studies have clearly shown that it differs from newly characterized M. quercophilus isolates. Subsequent analysis of laccase genes revealed that C30 sequences are similar to laccase encoding sequences from organisms belonging to the polyporoid clade. Comparison of sequences of the C30 ITS regions, including 5.8S rDNA, with those found in databanks confirmed that C30 is not a Marasmius. Finally, 25S rDNA analysis revealed that C30 is closely related to the Coriolaceae and, in particular, to Trametes trogii.

Vanillin production by recombinant strains of Escherichia coli

Vanillin production from ferulate was studied using different recombinant strains of Escherichia coli. To prevent the occurrence of aerobic conditions and then possible product oxidation, tests were performed in Erlenmeyer flasks under mild mixing (150 rpm). Among other transformants, E. coli JM109(pBB1) appeared to be the best vanillin producer, being able to convert no less than 95% of starting ferulate to the product within 1h. This yield decreased down to 72% after 72h, likely because of a non-specific oxidase activity responsible for vanillin oxidation to vanillate.

A produção de vanilina a partir de ácido ferúlico foi estudada utilizando-se diferentes linhagens recombinantes de Escherichia coli. Para prevenir a ocorrência de condições de aerobiose e a possível oxidação do produto, os ensaios foram realizados em frascos Erlenmeyer sob agitação moderada (150 rpm). E. coli JM109 (pBBI) mostrou-se o melhor produtor de vanilina entre os demais agentes transformantes, sendo capaz de converter 95% do ácido ferúlico inicial em produto após 1h, rendimento este que decresceu para 72% após 72h, provavelmente devido à atividade de uma oxidase não-específica responsável pela oxidação de vanilina a ácido vanílico.

Vanillin production by recombinant strains of Escherichia coli

Vanillin production from ferulate was studied using different recombinant strains of Escherichia coli. To prevent the occurrence of aerobic conditions and then possible product oxidation, tests were performed in Erlenmeyer flasks under mild mixing (150 rpm). Among other transformants, E. coli JM109(pBB1) appeared to be the best vanillin producer, being able to convert no less than 95% of starting ferulate to the product within 1h. This yield decreased down to 72% after 72h, likely because of a non-specific oxidase activity responsible for vanillin oxidation to vanillate.

A produção de vanilina a partir de ácido ferúlico foi estudada utilizando-se diferentes linhagens recombinantes de Escherichia coli. Para prevenir a ocorrência de condições de aerobiose e a possível oxidação do produto, os ensaios foram realizados em frascos Erlenmeyer sob agitação moderada (150 rpm). E. coli JM109 (pBBI) mostrou-se o melhor produtor de vanilina entre os demais agentes transformantes, sendo capaz de converter 95% do ácido ferúlico inicial em produto após 1h, rendimento este que decresceu para 72% após 72h, provavelmente devido à atividade de uma oxidase não-específica responsável pela oxidação de vanilina a ácido vanílico.