Maria Catarina Megumi Kasuya

Efeitos de diferentes formulações comerciais de glyphosate sobre estirpes de Bradyrhizobium

O objetivo deste trabalho foi avaliar efeitos de formulacoes comerciais de glyphosate sobre estirpes de Bradyrhizobium, em condicoes de laboratorio. As formulacoes foram aplicadas na concentracao de 43,2 µg L-1 do equivalente acido. As bacterias foram inoculadas em meio de cultura a base de manitol e extrato de levedura (YM). O efeito do herbicida no crescimento das estirpes de Bradyrhizobium foi avaliado mediante leitura da densidade otica em espectrofotometro. Avaliou-se o crescimento das estirpes de B. japonicum SEMIA 5079 e de B. elkanii SEMIA 5019 e SEMIA 587 sob efeito de nove formulacoes de glyphosate: Zapp Qiâ, Roundupâ, Roundup Multiacaoâ, Roundup Transorbâ, Roundup WGâ, Tropâ, Agrisatoâ, glyphosate tecnico [padrao de N-(phosphonomethyl) glycina] e controle sem adicao de herbicida (testemunha para as estirpes). Foram utilizadas seis repeticoes. Confeccionaram-se curvas de crescimento para cada estirpe. Pelos resultados, pode-se observar que todas as formulacoes de glyphosate causaram efeitos diferenciados sobre as estirpes de Bradyrhizobium SEMIA 5019, SEMIA 5079 e SEMIA 587. Constatou-se que a formulacao Zapp Qi foi a menos toxica as estirpes de Bradyrhizobium avaliadas. A maior toxicidade foi observada para Roundup Transorb, que provocou reducoes no crescimento acima de 94% para todas as estirpes de Bradyrhizobium estudadas. Nao se observou correlacao entre o tipo de sal - isopropilamina, amonio ou potassico, presentes na formulacao herbicida - e o grau de inibicao no crescimento das estirpes. SEMIA 587 foi a estirpe menos tolerante a maioria das formulacoes testadas, porem SEMIA 5019 foi a mais sensivel ao glyphosate padrao, sem adicao de sais ou de outros aditivos.

Polymorphism in the internal transcribed spacer (ITS) of the ribosomal DNA of 26 isolates of ectomycorrhizal fungi

Inter- and intraspecific variation among 26 isolates of ectomycorrhizal fungi belonging to 8 genera and 19 species were evaluated by analysis of the internal transcribed sequence (ITS) of the rDNA region using restriction fragment length polymorphism (RFLP). The ITS region was first amplified by polymerase chain reaction (PCR) with specific primers and then cleaved with different restriction enzymes. Amplification products, which ranged between 560 and 750 base pairs (bp), were obtained for all the isolates analyzed. The degree of polymorphism observed did not allow proper identification of most of the isolates. Cleavage of amplified fragments with the restriction enzymes Alu I, Hae III, Hinf I, and Hpa II revealed extensive polymorphism. All eight genera and most species presented specific restriction patterns. Species not identifiable by a specific pattern belonged to two genera: Rhizopogon (R. nigrescens, R. reaii, R. roseolus, R. rubescens and Rhizopogon sp.), and Laccaria (L. bicolor and L. amethystea). Our data confirm the potential of ITS region PCR-RFLP for the molecular characterization of ectomycorrhizal fungi and their identification and monitoring in artificial inoculation programs.

Epulorhiza epiphytica sp. nov. isolated from mycorrhizal roots of epiphytic orchids in Brazil

Abstract Epulorhiza epiphytica sp. nov. isolated from the roots of two Brazilian native epiphytic orchid species is described. In culture, it differs from the known species of Epulorhiza in the minute size of monilioid cells with foveate surfaces. This is the first report of an orchid mycorrhizal fungus from Brazil.

Vesicular-arbuscular-/ecto-mycorrhiza succession in seedlings of. Eucalyptus spp.

The occurrence of vesicular-arbuscular mycorrhizae (AM) and ectomycorrhizae (ECM) in the same root system was observed when species of Eucalyptus urophylla S.T. Blake, E. citriodora Hook f., E. grandis W. Hill ex Maiden, E. cloeziana F. Muell. and E. camaldulensis Dehnh were simultaneously inoculated with Glomus etunicatum Becker & Gederman and Pisolithus tinctorius (Per.) Cocker & Couch, isolate Pt 90A. The succession between the two fungi was observed. In general ectomycorrhizal colonization increased followed by a decrease in AM. Pisolithus tinctorius was favored in simultaneous inoculation with G. etunicatum, and the positive effect of the simultaneous inoculation of both fungi in the percent colonization by the AM fungus occurred up to 60 days after inoculation. After 120 days, colonization of roots by G. etunicatum decreased in the presence of P. tinctorius. When inoculated simultaneously, the proportion of AM and ECM varied with evaluation time, while the combined percentage of mycorrhizal roots approached the maximum and remained more or less constant after 60 days, suggesting that there could be competition between the fungi for limiting substrate. The maximum percent mycorrhizal colonization varied with Eucalyptus species and the highest value was observed for E. camaldulensis, followed in order by E. citriodora, E. urophylla, E. grandis and E. cloeziana.

Enrichment of mushrooms: An interesting strategy for the acquisition of lithium

The capability of Pleurotus ostreatus mushroom to accumulate lithium (Li) and the accessibility of this Li compared with lithium carbonate (Li(2)CO(3)), often used as psychiatric medicine, were investigated. Mushrooms were produced on a substrate-based on coffee husk, with different added concentrations of lithium chloride (LiCl). Biological efficiency (BE), the crude protein content, the concentration of Li and other elements present in mushrooms were determined. The sequential extraction and in vitro test were used to verify the accessibility and the degree of solubility of this element. Li concentration in mushrooms was directly influenced by increasing LiCl concentration in the substrate (P<0.05). The BE was not affected by different concentrations of LiCl. Li present in enriched mushrooms showed greater accessibility than in Li(2)CO(3). Therefore, P. ostreatus mushrooms, enriched with lithium can be an alternative source of Li, as well as being a food with high nutritional value.

Molecular characterization of Pisolithus spp. isolates by rDNA PCR-RFLP

Abstract Variation within ribosomal DNA (rDNA) genes of 19 isolates of Pisolithus from different geographic origins and hosts was examined by polymerase chain reaction (PCR) coupled with restriction fragment length polymorphism (RFLP) analysis. The primers utilized amplify rDNA regions in a wide range of fungi. One amplified region includes the internal transcribed spacer (ITS), which has a low degree of conservation. The ITS amplification products (640–750 bp) were digested with a variety of restriction endonucleases. Cluster analysis based on the restriction fragments grouped the isolates into three distinct groups: group I contained isolates collected in the northern hemisphere, except Pt 1, group II contained those collected in Brazil and group III contained isolate Pt 1. Additional analysis of other rDNA regions, IGS, 17 S and 25 S rDNA, resulted in similar groups. The data suggest that the taxonomy and systematics of this ectomycorrhizal fungus should be revised.

In vivo bioavailability of selenium in enriched Pleurotus ostreatus mushrooms

The in vivo bioavailability of Se was investigated in enriched Pleurotus ostreatus mushrooms. A bioavailability study was performed using 64 Wistar male rats separated in 8 groups and fed with different diets: without Se, with mushrooms without Se, with enriched mushrooms containing 0.15, 0.30 or 0.45 mg kg(-1) Se and a normal diet containing 0.15 mg kg(-1) of Se using sodium selenate. The experiment was performed in two periods: depletion (14 days) and repletion (21 days), according to the Association of Official Analytical Chemists. After five weeks, the rats were sacrificed under carbon dioxide, and blood was drawn by heart puncture. Blood plasma was separated by centrifugation. The total Se concentration in the plasma of rats fed with enriched mushrooms was higher than in rats fed with a normal diet containing sodium selenate. The plasma protein profiles were obtained using size exclusion chromatography (SEC) and UV detectors. Aliquots of effluents (0.5 mL per minute) were collected throughout in the end of the chromatographic column. However, Se was determined by graphite furnace atomic absorption spectrometry (GF AAS) only in the aliquots where proteins were detected by SEC-UV. The plasma protein profile of rats fed with different diets was similar. The highest Se concentration was observed in a peptide presenting 8 kDa. Furthermore, the higher Se concentration in this peptide was obtained for rats fed with a diet using enriched mushrooms (7 μg L(-1) Se) compared to other diets (2-5 μg L(-1) Se). These results showed that Se-enriched mushrooms can be considered as an alternative Se food source for humans, due to their high bioavailability.

A pH signaling mechanism involved in the spatial distribution of calcium and anion fluxes in ectomycorrhizal roots.

Mycorrhization is a typical example of a host-pathogen symbiotic interaction where the pathogen cell biology and the host immune response coevolved several functional links. Here, the role played by ion fluxes across the root concerning nutrient uptake, osmoregulation, growth and signaling events is addressed. An ion-selective vibrating probe system was used to determine the net fluxes of protons (H(+)), calcium (Ca(2+)) and anions (A(-)) along nonmycorrhizal and ectomycorrhizal (ECM) roots of Eucalyptus globulus colonized by Pisolithus sp. These data show that, from five root zones analyzed, the main effect of fungal colonization was localized to the elongation zone. Here, strong changes in ion dynamics and rhizosphere acidification capacity were observed. Additionally, ion fluxes exhibited periodic fluctuations. To verify whether these fluctuations corresponded to sustained oscillations, continuous wavelet time spectrum analysis was applied and it was determined that H(+) and A(-) fluxes from ECM roots had longer periods than nonmycorrhizal roots. By contrast, Ca(2+) oscillations were completely abolished following fungal interaction. These results are interpreted in the light of a working model in which nutrient uptake and stimulation of growth are mediated by ECM fungi and may be pH-dependent. Furthermore, the variations detected in ECM roots for H(+) and A(-) fluxes suggest a main contribution from the plant, while the results obtained for Ca(2+) point to a significant involvement of the fungus.

Degradation of Oxo-Biodegradable Plastic by Pleurotus ostreatus

Growing concerns regarding the impact of the accumulation of plastic waste over several decades on the environmental have led to the development of biodegradable plastic. These plastics can be degraded by microorganisms and absorbed by the environment and are therefore gaining public support as a possible alternative to petroleum-derived plastics. Among the developed biodegradable plastics, oxo-biodegradable polymers have been used to produce plastic bags. Exposure of this waste plastic to ultraviolet light (UV) or heat can lead to breakage of the polymer chains in the plastic, and the resulting compounds are easily degraded by microorganisms. However, few studies have characterized the microbial degradation of oxo-biodegradable plastics. In this study, we tested the capability of Pleurotus ostreatus to degrade oxo-biodegradable (D2W) plastic without prior physical treatment, such as exposure to UV or thermal heating. After 45 d of incubation in substrate-containing plastic bags, the oxo-biodegradable plastic, which is commonly used in supermarkets, developed cracks and small holes in the plastic surface as a result of the formation of hydroxyl groups and carbon-oxygen bonds. These alterations may be due to laccase activity. Furthermore, we observed the degradation of the dye found in these bags as well as mushroom formation. Thus, P. ostreatus degrades oxo-biodegradable plastics and produces mushrooms using this plastic as substrate.

Diversity of Fungi Associated with Atta bisphaerica (Hymenoptera: Formicidae): The Activity of Aspergillus ochraceus and Beauveria bassiana

The grass-cutting ant Atta bisphaerica is one of the most serious pests in several pastures and crops in Brazil. Fungal diseases are a constant threat to these large societies composed of millions of closely related individuals. We investigated the occurrence of filamentous fungi associated with the ant A. bisphaerica in a pasture area of Vicosa, Minas Gerais State, Brazil. Several fungi species were isolated from forager ants, and two of them, known as entomopathogenic, Beauveria bassiana and Aspergillus ochraceus, were tested against worker ants in the laboratory. The two species were highly virulent, achieving 50 percent worker mortality within 4-5 days. It is the first time A. ochraceus, a commonly found fungal species, is reported to infect Atta species at a high prevalence. Possible uses for the fungus within biological control are discussed.