This study used a multiphasic approach, characterized by the simultaneous use of culture-dependent and culture-independent methods, to investigate endophytic bacterial communities in strawberry (Fragaria ananassa) fruit. A total of 92 bacterial endophytes were isolated and initially grouped by their repetitive extragenic palindromic (rep)-PCR banding pattern and biochemical features. Phylogenetic analysis of the 16S rRNA gene sequences of 45 representatives showed that the isolates belonged to the species Bacillus subtilis (eight isolates), Bacillus sp. (seven isolates), Enterobacter sp. (seven isolates), Enterobacter ludwigii (six isolates), Lactobacillus plantarum (six isolates), Pseudomonas sp. (five isolates), Pantoea punctata (three isolates), and Curtobacterium citreum (three isolates). Nucleic acids were extracted from the strawberry fruit and subjected to 16S rRNA gene directed polymerase chain reaction denaturing gradient gel electrophoresis (16S rRNA PCR-DGGE). The species B. subtilis, Enterobacter sp., and Pseudomonas sp. were detected both by isolation and DGGE. The DGGE fingerprints of total bacterial DNA did not exhibit bands corresponding to several of the representative species isolated in the extinction dilution (L. plantarum, C. citreum, and P. punctata). In contrast, bands in the DGGE profile that were identified as relatives of Arthrobacter sp. and one uncultivable Erythrobacter sp. were not recovered by cultivation techniques. After isolation, the nitrogen fixation ability and the in vitro production of indole-3-acetic acid (IAA) equivalents and siderophores were evaluated. A high percentage of isolates were found to possess the ability to produce siderophores and IAA equivalents; however, only a few isolates belonging to the genera Pseudomonas and Enterobacter showed the ability to fix nitrogen. Plant growth promotion was evaluated under greenhouse conditions and revealed the ability of the Bacillus strains to enhance the number of leaves, shoot length, root dry weight, and shoot dry weight. The activity of the bacterial isolate identified as B. subtilis NA-108 exerted the greatest influence on strawberry growth and showed a 42.8% increase in number of leaves, 15.26% for high shoot, 43.5% increase in root dry weight, and a 77% increase in shoot dry weight when compared with untreated controls.

Composting is the process of natural degradation of organic matter carried out by environmental microorganisms whose metabolic activities cause the mineralization and partial humification of substances in the pile. This compost can be beneficially applied to the soil as organic fertilizer in horticulture and agriculture. The number of studies involving microbial inoculants has been growing, and they aim to improve processes such as composting. However, the behavior of these inoculants and other microorganisms during the composting process have not yet been described. In this context, this work aimed to investigate the effects of using a microbial inoculum that can improve the composting process and to follow the bacterial population dynamics throughout the process using the high-resolution melt (HRM) technique. To do so, we analysed four compost piles inoculated with Bacillus cereus, Bacillus megaterium, B. cereus + B. megaterium and a control with no inoculum. The analyses were carried out using samples collected at different stages of the process (5th to 110th days). The results showed that the bacterial inocula influenced the process of composting, altering the breakdown of cellulose and hemicelluloses and causing alterations to the temperature and nitrogen levels throughout the composting process. The use of a universal primer (rDNA 16S) allowed to follow the microbial succession during the process. However, the design of a specific primer is necessary to follow the inoculum throughout the composting process with more accuracy.Index terms: High Resolution Melt (HRM); bacteria; microbial ecology. RESUMOA compostagem é um processo de degradação natural da matéria orgânica realizado por microrganismos presentes no ambiente, levando a mineralização e humificação parcial das substâncias presentes na pilha, esse composto formado pode ser beneficamente aplicado ao solo como fertilizante orgânico na horticultura e agricultura. O número de estudos envolvendo inoculantes microbianos é crescente, os quais tem por objetivo a otimização de processos de compostagem. Contudo, o comportamento desses inoculantes e da microbiota ao longo do processo não tem sido caracterizado. Nesse contexto, este trabalho foi realizado com o objetivo de avaliar o efeito da utilização de um inóculo bacteriano que promova melhorias no processo de compostagem, bem como o de acompanhar a dinâmica populacional bacteriana ao longo de todo o processo através da técnica de High Resolution Melt (HRM). Para isso foram analisados quatro pilhas de compostagem inoculadas com Bacillus cereus, Bacillus megaterium, B. cereus + B. megaterium e o controle sem adição de inóculo. Foram realizadas análises químicas e moleculares (HRM) das amostras coletadas em diferentes períodos da compostagem (5º ao 110º dias). Os resultados mostraram que os inóculos bacterianos influenciaram no processo de compostagem com alteração na degradação de celulose, hemicelulose bem como alteração da temperatura e níveis de nitrogênio ao longo da compostagem. A utilização de ...

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The composition and genetic diversity of fungal populations during phase II of compost production for the cultivation of Agaricus subrufescens was determined using culture-dependent and -independent methods on days 3, 6, 10, 12, and 14 of phase II composting. The isolates were morphologically characterized and subsequently analyzed using repetitive extragenic palindromic sequences (rep-PCR), and the intergenic region was sequenced to genetically identify the isolates. Changes on in the filamentous fungi population were analyzed using denaturing gradient gel electrophoresis (DGGE), and the resulting bands were sequenced. The population did not significantly change from day 3 to 10 (2.55 x 10(5) -6 x 10(5) CFU g(-1)), and maximum counts on day 14 of phase II composting (6.92 log CFU g(-1)). In the morphological characterization, Scytalidium thermophilum, Thermomyces lanuginosus, and Thermomyces ibadanensis were the most abundant identified species. The 26 most abundant isolates identified by morphological analysis were characterized using rep-PCR. A significant amount of genetic diversity was detected among the isolates of all three studied species. Based on the DGGE analysis, the diversity of the fungi was reduced during phase II composting, and S. thermophilum was the predominant species identified throughout the entire process. Thus, this study presents the first report of the involvement of T. ibadanensis in the production of compost for Agaricus mushroom cultivation.

The production of tomato seedlings is conducted on commercial substrates with adequate properties for the good formation of the aerial part and root. The Spent Mushroom Substrate, or SMS, presents advantages over commercial substrates regarding the quality of the vegetable seedlings, which may be provided by the presence of a rich microbiota, bringing higher balance and competition with pathogenic microorganisms, in addition to the biological control of pathogens and nematodes. It is important to know the microbiota present in this material and its relation to the plant, in order for this association to occur in the best manner possible. This work had the objective of identifying the microbiota present in the rhizosphere of tomato seedlings produced in SMS of Agaricus subrufescens and Agaricus bisporus mushrooms, added or not with commercial substrate. The microbiota was analyzed by DGGE and the representative samples were sequenced in order to identify the species. Among the eukaryotes, the Chaetomium globosum, Arthrobotrys amerospora species were predominant in the A. subrufescens SMS, and Geastrum floriforme, Chrysosporium chiropterum, Pleurotus sp., and Trichosporon sp. in the A. bisporus SMS. Prokaryotes are the most diverse group, suggesting a large number of species not yet classified or which sequences are not yet available.