Eucalyptus Essential Oil As Bio-stimulator Of The Growth Of In Vitro Ectomycorrhizal Fungi [Óleo Essencial De Eucalipto Como Bioestimulador Do Crescimento De Fungos Ectomicorrízicos In Vitro]

The secondary metabolites extracted from forest species known as micossimbiontes can stimulate the growth of in vitro ectomycorrhizal isolates. We determined the effect of concentration of Eucalyptus grandis essential oil in stimulating growth of the ectomycorrhizal Pisolithus sp. (UFSC Pt 24 and UFSC Pt 188), Pisolithus microcarpus (UFSC Pt 116) Chondrogaster angustiporus (UFSC Ch 163), Scleroderma citrinum (UFSC Sc 124) and Suillus sp. (UFSM RA RA UFSM 2.2 and 2.8) in liquid culture medium. After a period of 25 days of incubation, we evaluated the morphology and growth of the isolates. The addition of essential oil at concentrations from 20 to 30 μL L-1 promoted mycelial growth in vitro in isolated Pt 24 UFSC, UFSC Pt 116, Ch 163 UFSC, UFSC Sc 124, UFSM RA RA UFSM 2.2 and 2.8. The addition of essential oil at a concentration of 20 μL L-1 resulted in an increase in the diameter and branching hyphae of isolates UFSC Pt 116 and Pt 24 UFSC. 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Efeitos de diferentes formulações comerciais de glyphosate sobre estirpes de Bradyrhizobium Effects of different glyphosate commercial formulations on Bradyrhizobium strains

O objetivo deste trabalho foi avaliar efeitos de formulações comerciais de glyphosate sobre estirpes de Bradyrhizobium, em condições de laboratório. As formulações foram aplicadas na concentração de 43,2 µg L-1 do equivalente ácido. As bactérias foram inoculadas em meio de cultura à base de manitol e extrato de levedura (YM). O efeito do herbicida no crescimento das estirpes de Bradyrhizobium foi avaliado mediante leitura da densidade ótica em espectrofotômetro. Avaliou-se o crescimento das estirpes de B. japonicum SEMIA 5079 e de B. elkanii SEMIA 5019 e SEMIA 587 sob efeito de nove formulações de glyphosate: Zapp Qi®, Roundup®, Roundup Multiação®, Roundup Transorb®, Roundup WG®, Trop®, Agrisato®, glyphosate técnico [padrão de N-(phosphonomethyl) glycina] e controle sem adição de herbicida (testemunha para as estirpes). Foram utilizadas seis repetições. Confeccionaram-se curvas de crescimento para cada estirpe. Pelos resultados, pôde-se observar que todas as formulações de glyphosate causaram efeitos diferenciados sobre as estirpes de Bradyrhizobium SEMIA 5019, SEMIA 5079 e SEMIA 587. Constatou-se que a formulação Zapp Qi foi a menos tóxica às estirpes de Bradyrhizobium avaliadas. A maior toxicidade foi observada para Roundup Transorb, que provocou reduções no crescimento acima de 94% para todas as estirpes de Bradyrhizobium estudadas. Não se observou correlação entre o tipo de sal - isopropilamina, amônio ou potássico, presentes na formulação herbicida - e o grau de inibição no crescimento das estirpes. SEMIA 587 foi a estirpe menos tolerante à maioria das formulações testadas, porém SEMIA 5019 foi a mais sensível ao glyphosate padrão, sem adição de sais ou de outros aditivos.<br>This work aimed to evaluate the effects of glyphosate commercial formulations on Bradyrhizobium strains under laboratory conditions. The formulations were applied in the concentration of 43.2 µg L-1 of the a.e. and the strains were inoculated in yeast extract manitol (YM). Herbicide effect on the growth of the Bradyrhizobium strains was assessed by optic density reading in a spectrophotometer. Twenty seven treatments arranged in a factorial design were evaluated and consisted of one strain of B. japonicum: SEMIA 5079; and two strains of B. elkanii: SEMIA 5019 and SEMIA 587, under the effect of nine glyphosate formulations: Zapp QI®, Roundup®, Roundup Multiação®, Roundup Transorb®, Roundup WG®, Trop®, Agrisato®, technical glyphosate [N-(phosphonomethyl) glycine] and control without herbicide addition (as the strain control treatment), with six replications. A growth curve was established for each strain. It could be observed that the different glyphosate formulations Zapp Qi, Roundup, Roundup Multiação, Roundup transorb, Roundup WG, Trop and Agrisato caused differentiated effects on the strains of Bradyrhizobium SEMIA 5019, SEMIA 5079 and SEMIA 587. It was verified that the Zapp Qi formulation was the least toxic to the strains. The highest toxicity was observed for Roundup Transorb, which reduced growth over 94% for all the strains assessed. Correlation was not observed among the type of salt, isopropylamine, ammonium or potassic, present in the formulation herbicides, and the toxicity degree to the strains. The strain SEMIA 587 was the least tolerant to most formulations while SEMIA 5019 was the most sensitive to the control treatment N- (phosphonomethyl) glycine, without salts or other additives

Molecular analysis of the bacterial diversity in a specialized consortium for diesel oil degradation Análise molecular da diversidade bacteriana de um consórcio degradador de óleo diesel

Diesel oil is a compound derived from petroleum, consisting primarily of hydrocarbons. Poor conditions in transportation and storage of this product can contribute significantly to accidental spills causing serious ecological problems in soil and water and affecting the diversity of the microbial environment. The cloning and sequencing of the 16S rRNA gene is one of the molecular techniques that allows estimation and comparison of the microbial diversity in different environmental samples. The aim of this work was to estimate the diversity of microorganisms from the Bacteria domain in a consortium specialized in diesel oil degradation through partial sequencing of the 16S rRNA gene. After the extraction of DNA metagenomics, the material was amplified by PCR reaction using specific oligonucleotide primers for the 16S rRNA gene. The PCR products were cloned into a pGEM-T-Easy vector (Promega), and Escherichia coli was used as the host cell for recombinant DNAs. The partial clone sequencing was obtained using universal oligonucleotide primers from the vector. The genetic library obtained generated 431 clones. All the sequenced clones presented similarity to phylum Proteobacteria, with Gammaproteobacteria the most present group (49.8 % of the clones), followed by Alphaproteobacteira (44.8 %) and Betaproteobacteria (5.4 %). The Pseudomonas genus was the most abundant in the metagenomic library, followed by the Parvibaculum and the Sphingobium genus, respectively. After partial sequencing of the 16S rRNA, the diversity of the bacterial consortium was estimated using DOTUR software. When comparing these sequences to the database from the National Center for Biotechnology Information (NCBI), a strong correlation was found between the data generated by the software used and the data deposited in NCBI.<br>O óleo diesel é um composto derivado do petróleo, constituído basicamente por hidrocarbonetos. Condições precárias no processo de transporte e armazenagem desse produto contribuem significativamente para derrames acidentais, ocasionando sérios problemas ecológicos no solo e água, alterando assim toda a diversidade microbiológica do ambiente. A estratégia de clonagem e sequenciamento do gene 16S rRNA é uma das técnicas moleculares que permitem estimar e comparar a diversidade microbiana de diferentes amostras ambientais, sejam elas impactadas ou não. O objetivo deste trabalho foi estimar a diversidade de microrganismos pertencentes ao domínio Bacteria em um consórcio degradador de óleo diesel por meio de sequenciamento parcial do gene 16S rRNA. Após extração do DNA metagenômico, o material foi amplificado por reação de PCR com oligonucleotídeos iniciadores específicos para o gene 16S rRNA. Os produtos da reação de PCR foram clonados em vetor pGEM T Easy (Promega) e transformados em células competentes de Escherichia coli. O sequenciamento parcial dos clones foi feito com oligonucleotídeos universais do vetor. A biblioteca obtida gerou 431 clones. Todos os clones mostraram similaridade com o filo Proteobacteria, onde as Gammaproteobacteria compreenderam o grupo de maior representatividade, com 49,8 % dos clones, seguida das Alphaproteobacteira, com 44,8 %, e das Betaproteobacteria, com 5,4 %. O gênero Pseudomonas destacou-se como representante com maior frequência de clones na biblioteca, seguido pelos gêneros Parvibaculum e Sphingobium. Após o sequenciamento parcial do gene 16S rRNA, a diversidade bacteriana do consórcio foi estimada utilizando-se o software DOTUR. Essas sequências, quando comparadas com as do banco do National Center for Biotechnology Information (NCBI), mostraram grande correlação entre os dados gerados pelo software utilizado e aqueles depositados no NCBI

Changes in bacterial community of anthracene bioremediation in municipal solid waste composting soil*

Polycyclic aromatic hydrocarbons (PAHs) are common contaminants in a municipal solid waste (MSW) composting site. Knowledge of changes in microbial structure is useful to identify particular PAH degraders. However, the microbial community in the MSW composting soil and its change associated with prolonged exposure to PAHs and subsequent biodegradation remain largely unknown. In this study, anthracene was selected as a model compound. The bacterial community structure was investigated using terminal restriction fragment length polymorphism (TRFLP) and 16S rRNA gene clone library analysis. The two bimolecular tools revealed a large shift of bacterial community structure after anthracene amendment and subsequent biodegradation. Genera Methylophilus, Mesorhizobium, and Terrimonas had potential links to anthracene biodegradation, suggesting a consortium playing an active role