Microbiologia
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Item Production of pectin lyase by Penicillium griseoroseum as a function of the inoculum and culture conditions(World Journal of Microbiology and Biotechnology, 1992-10-09) Brumano, M. H. N.; Coelho, J. L. C.; Araújo, E. F.; Silva, D. O.Optimum activity of an extracellular pectin lyase produced by Penicillium griseoroseum in submerged culture was after 120 h using 0.1% (w/v) citrus pectin as substrate. Sucrose at 0.1% (w/v) stimulated enzyme production and citrus pectin gave the highest activity of enzyme per unit growth.Item Occurrence and types of ectomycorrhizae present in seedlings of Picea glehnii in a natural forest in Hokkaido(Mycoscience, 1995-10) Kasuya, Maria Catarina Megumi; Igarashi, Tsuneo; Masato ShibuyaPicea glehnii seedlings whose ages ranged from 2 to 5 years and which were growing in aP. glehnii-Abies sachalinensis natural forest where the surface had been scarified 6 years before, were assessed for the occurrence and types of ectomycorrhizae. Based on the macro- and microscopic characteristics, 34 types of ectomycorrhizae were classified. The basidiomycetes were clearly dominant and it was common to observe more than one type of mycorrhiza occurring in the same root tip. The diversity of mycorrhizal types and the presence of at least one type of ectomycorrhiza in a single seedling, observed in this work, indicate a very dynamic system of mycorrhizal formation inP. glehnii seedlings, which allows them to survive and grow in a forest surface subjected to soil scarification.Item Production and regeneration of protoplasts from the mycorrhizal fungus Suillus granulatus(World Journal of Microbiology and Biotechnology, 1996-04-11) Dias, E. S.; Araújo, E. F.; Guimarães, W. V.; Muchovej, R. M. C.Experiments were performed with the mycorrhizal fungus Suillus granulatus to define the parameters for production and regeneration of protoplasts. Protoplasts were released at frequencies between 1 and 3×107/ml from mycelium 3 to 7 days old. The best osmotic stabilizer for protoplast release was MgSO4 (0.7 m). To optimize protoplast release and regeneration an enzyme (Novozym 234) concentration 1.7 mg/ml was chosen, with a digestion time of 1 to 2 h. Regenerated colonies formed mycorrhizae within 60 days after inoculation in Pinus caribaea var. hondurensis seedlings.Item Genetic diversity of the ectomycorrhizal fungus Pisolithus tinctorius based on RAPD-PCR analysis(Mycorrhiza, 1997-10-03) Junghans, D. T.; Gomes, E. A.; Guimarães, W. V.; Barros, E. G.; Araújo, E. F.Twenty Pisolithus tinctorius isolates from different geographic locations and different hosts were characterized by the random amplified polymorphic DNA technique. Thirteen arbitrary primers generated 87 DNA fragments, all of them polymorphic. These data were used to calculate genetic distances among the isolates. The pairwise genetic distances ranged from 1 to 100%, with an average of 58.7%. Cluster analysis based on the amplified fragments grouped the isolates according to their host and geographical origins. Group I contained isolates collected in Brazil and group II those collected in the Northern Hemisphere. In addition to the diversity seen at the molecular level, the isolates also showed host specificity. Greenhouse experiments demonstrated that isolates from the Northern Hemisphere colonized mainly Pinus whereas isolates from Brazil colonized only Eucalyptus. The molecular data suggest that the Pisolithus tinctorius isolates analyzed belong to two distinct groups. The data also suggest new guidelines for future investigations on the taxonomy and systematic of this important fungus species. Furthermore, these results support future experiments aimed at the selection and development of improved isolates of P. tinctorius.Item Estudo da atividade antibacteriana de plantas medicinais, aromáticas e corantes naturais(Revista Brasileira de Farmacognosia, 1998) Bara, Maria Teresa Freitas; Vanetti, Maria Cristina DantasO emprego de recursos naturais como mecanismo auxiliar no tratamento de distintas patologias tem ressurgido, com ênfase, nos útimos tempos. Tendo em vista esta situação, este estudo teve por finalidade investigar a atividade antimicrobiana de algumas plantas medicinais e aromáticas bastantes comuns na cultura brasileira. Foi constatado que, de modo particular, o alecrim-pimenta (Lippia sidoides) levou a uma considerável inibição do crescimento bacteriano. Tambêm plantas como alecrim (Rosmarinus officinalis), cravo (Sysygium aromaticum), noz-moscada (Myristica fragrans), pimenta-da-jamaica (Pimenta dioica) e sálvia (Salvia officinalis) demonstraram constituir fontes eficientes de compostos bioativos antibacterianos.Item Variação sazonal da colonização de raízes de clones de híbridos de eucalipto por fungos micorrízicos no estado do Espírito Santo(Revista Brasileira de Ciência do Solo, 1998-07) Grazziotti, P. H.; Barros, N. F.; Borges, A. C.; Neves, J. C. L.; Fonseca, S.A colonização de raízes de clones de três híbridos de Eucalyptus grandis x E. urophylla por fungos micorrízicos foi estudada no campo em diferentes condições de solo. A colonização por fungos ectomicorrízicos e micorrízicos arbusculares foi avaliada em três locais do Espírito Santo, em quatro épocas de amostragem (julho e outubro de 1992 e janeiro e abril de 1993). A unidade experimental foi constituída por três árvores com cinco anos de idade. A colonização micorrízica mostrou-se dependente da área, da época de amostragem e dos clones. A ocorrência de ectomicorrizas foi maior no verão, quando se registraram as maiores temperaturas. A intensidade de colonização por fungos micorrízicos arbusculares foi semelhante entre as áreas e não apresentou variação consistente em função da época. O número de esporos de fungos micorrízicos arbusculares por grama de solo variou de acordo com a área e época de amostragem.Item Characterization of an Aspergillus nidulans mutant with abnormal distribution of nuclei in hyphae, metulae, phialides and conidia(FEMS Microbiology Letters, 1998-07-10) Queiroz, Marisa Vieira de; Azevedo, João Lúcio deThe V10 deteriorated variant of Aspergillus nidulans has hyphae, metulae, phialides and conidia with abnormal nuclear distributions. The alterations observed were: increase in the number of nuclei in hyphae, metulae and phialides, presence of anucleate, uninucleate and multinucleate conidia, abnormal vegetative growth and defective conidiation. When 0.5 M NaCl was added to the medium, an increase in the number of conidia was observed but their morphology and number of nuclei were not modified. The gene responsible for these alterations was named anuA1. The anuA1 gene is located on linkage group VII and is possibly involved in nuclear migration to hyphae, metulae, phialides and conidia. z 1998 Federation of European Microbiological SocietiesItem Fermentation of maltose and starch by Klebsiella oxytoca P2(Biotechnology Letters, 1998-12) Santos, Vera L. dos; Guimarães, Walter V.; Barros, Everaldo G. de; Araújo, Elza F.Klebsiella oxytoca P2, which has genes from Zymomonas mobilis encoding the alcohol dehydrogenase and pyruvate decarboxylase integrated in its chromosome, fermented 50 g maltose/l to 25.4 g of ethanol/l. It also fermented 10, 20 and 40 g starch/l yielding 4, 8.4, and 17.7 g ethanol/l, respectively, representing 72, 75 and 78% of the theoretical yield.Item Cloning and characterization of a gene encoding the endopolygalacturonase of Penicillium griseoroseum(Biotechnology Letters, 1999-05) Ribon, A. O. B.; Coelho, J. L. C.; Barros, E. G. de; Araújo, E. F.A conserved region of a polygalacturonase (PG) gene from Penicillium griseoroseum was PCR amplified and used to screen a genomic library from this fungus. The nucleotide sequence of the isolated clone (pggI) consisted of 1497 bp, including a coding region of 1251 bp. This region potentially encodes a protein of 376 amino acids, and is interrupted by two introns. Extensive homology was observed between this protein and several fungal endopolygalacturonases. DNA hybridization analyses revealed that there is a low copy number of pggI in the P. griseoroseum genome, probably one or two copies.Item Utilização de características microbiológicas na avaliação da degradação ou recuperação de uma área sob mineração de ferro(Revista Brasileira de Ciência do Solo, 2000-05) Trindade, A. V.; Grazziotti, P. H.; Tótola, M. R.O estado de degradação e, ou, recuperação de diferentes sítios de uma área de mineração de ferro no município de Mariana (MG) foi avaliado pela atividade microbiana, número de esporos de fungos micorrízicos arbusculares (FMA) e população total de fungos e bactérias. Os substratos presentes em cada sítio, caracterizados como Rejeito de flotação (Rj), Solo superficial (SS), Subsolo (Ss), Subsolo com vegetação (SsV), Área em revegetação (AR) e Mata secundária (MS), foram amostrados na profundidade de 10-25 cm, retirando-se uma amostra composta, formada por oito amostras simples. A atividade microbiana foi avaliada pela evolução de CO2 em amostras ao natural e com adição de sacarose ou sacarose + uréia. Fez-se também a determinação de características químicas e teor de umidade. A colonização do substrato SsV por gramíneas e plantas de candeia (Vanillosmopsis erythropappa Schult. Bip.) foi acompanhada pelo número de esporos de FMA, teor de carbono orgânico e atividade microbiana. Os esporos de FMA só foram observados em áreas com vegetação. O armazenamento da camada superficial de solo (sítio SS) reduziu os teores de carbono orgânico, nitrogênio total, número de esporos de FMA e atividade microbiana. Nos substratos resultantes da atividade mineradora, houve necessidade do fornecimento conjunto de carbono e nitrogênio para estimular a atividade microbiana. As características microbiológicas avaliadas apresentaram valores maiores no sítio de mata secundária do que nos demais amostrados, indicando que aqueles em recuperação não possuíam o mesmo equilíbrio biológico. As avaliações microbiológicas do solo foram capazes de distinguir os estados de perturbação ou recuperação das diferentes áreas estudadas.Item Electrophoretic characterization ofaspergillus nidulans strains with chromosomal duplications(Genetics and Molecular Biology, 2000-06) Queiroz, Marisa V. de; Pizzirani-Kleiner, Aline Aparecida; Azevedo, João LúcioPulsed-field gel electrophoresis was used to characterize strains ofAspergillus nidulans with a chromosomal duplication Dp(I-II). Morphologically deteriorated and improved variants of these strains were also analyzed. The electrophoretic karyotype demonstrated that in two duplicated strains (A and B) the 4.2 Mb band, which corresponds to chromosome II, was absent and a new band was observed. Hybridization studies using the uapA (chromosome I) and wA (chromosome II) genes demonstrated that the new band corresponded to chromosome II plus the duplicated segment of chromosome I. The size of the chromosomal duplication was approximately 1.0 Mb. Analysis of the chromosomal bands of a morphologically improved strain showed that the duplicated segment of chromosome I was completely lost. The morphologically deteriorated variants V9 and V17 had the same karyotype as the duplicated strains. However, the deteriorated variant V5 lost part of chromosome I and had a rearrangement involving chromosome V. This rearrangement may have resulted from the mutagenic treatment used to obtain the genetic markers. Pulsed-field gel electrophoresis was found to be an excellent tool for locating chromosomal rearrangements.Item Fermentation of sweet whey by recombinant Escherichia coli KO11(Brazilian Journal of Microbiology, 2000-07) Leite, Amarildo Ricardo; Guimarães, Walter Vieira; Araújo, Elza Fernandes de; Silva, Daison OlzanyThe production of ethanol from sweet whey using the recombinant Escherichia coli KO11, in batch fermentation, was tested. The maximum ethanol yield was reached after 96h, representing only 38% of the theoretical yield. The supplementation of whey with components of LB broth increased the maximum yield to 96% in 72h. The addition of 0.5% yeast extract to whey resulted in maximum yield of 74% at 36h and it increased to over 100% when yeast extract and trace metals solution (Fe++, Mn++ and Zn++) were added.Item Production of pectin lyase by Penicillium griseoroseum in bioreactors in the absence of inducer(Brazilian Journal of Microbiology, 2001-04) Passos, Flávia Maria Lopes; Passos, Frederico José Vieira; Silva, Daison Olzany; Piccoli-Valle, Roberta HilsdorfPenicillium griseoroseum was grown in bioreactors on mineral medium supplemented with yeast extract and sucrose. The influence of inoculum and carbon source concentrations, aeration and pH on pectin lyase (PL) production, as well as the capacity of P. griseoroseum to produce PL when grown on sugar cane syrup as carbon source were evaluated. Inoculum concentration did not influence PL production. Production was higher in non-aerated than in aerated medium. The best results were obtained using 60 mM sucrose at pH 6.3-7.2. Production using cane syrup 25% (v/v), without yeast extract supplement, was equal to that obtained under the conditions cited above.Item The high fermentative metabolism of Kluyveromyces marxianus UFV-3 relies on the increased expression of key lactose metabolic enzymes(Antonie van Leeuwenhoek, 2001-11-09) Diniz, Raphael H. S.; Silveira, Wendel B.; Fietto, Luciano G.; Passos, Flávia M. L.The aim of this work was to obtain insights about the factors that determine the lactose fermentative metabolism of Kluyveromyces marxianus UFV-3. K. marxianus UFV-3 and Kluyveromyces lactis JA6 were cultured in a minimal medium containing different lactose concentrations (ranging from 0.25 to 64 mmol l−1) under aerobic and hypoxic conditions to evaluate their growth kinetics, gene expression and enzymatic activity. The increase in lactose concentration and the decrease in oxygen level favoured ethanol yield for both yeasts but in K. marxianus UFV-3 the effect was more pronounced. Under hypoxic conditions, the activities of β-galactosidase and pyruvate decarboxylase from K. marxianus UFV-3 were significantly higher than those in K. lactis JA6. The expression of the LAC4 (β-galactosidase), RAG6 (pyruvate decarboxylase), GAL7 (galactose-1-phosphate uridylyltransferase) and GAL10 (epimerase) genes in K. marxianus UFV-3 was higher under hypoxic conditions than under aerobic conditions. The high expression of genes of the Leloir pathway, LAC4 and RAG6, associated with the high activity of β-galactosidase and pyruvate decarboxylase contribute to the high fermentative flux in K. marxianus UFV-3. These data on the fermentative metabolism of K. marxianus UFV-3 will be useful for optimising the conversion of cheese whey lactose to ethanol.Item Structural organization of polygalacturonase-encoding genes from Penicillium griseoroseum(Genetics and Molecular Biology, 2002) Ribon, Andréa O. B.; Queiroz, Marisa V.; Araújo, Elza F. deThe pectinolytic system of Penicillium griseoroseum has been studied as a model to investigate aspects of gene organization in filamentous fungi. Here we show that the endopolygalacturonase-coding genes previously isolated exist as single copies in the fungus genome. DNA blot analysis revealed the presence of corresponding genes in other Penicillium species, although only one or two genes were found in opposition to the endoPG gene family reported for other filamentous fungi. The nucleotide and amino acid sequences of Penicillium PG genes of retrieved from data banks were compared for intron length and number, codon usage, and consensus sequences for translation initiation sites. The introns are conserved in the same position, although there was no conservation of their nucleotide sequences. Other sequence features resemble those seen in Aspergillus and Neurospora genesItem Polymorphism in the internal transcribed spacer (ITS) of the ribosomal DNA of 26 isolates of ectomycorrhizal fungi(Genetics and Molecular Biology, 2002) Gomes, Eliane A.; Kasuya, Maria Catarina M.; Barros, Everaldo G. de; Borges, Arnaldo C.; Araújo, Elza F.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.Item Molecular characterization and evaluation of pectinase and cellulase production of Penicillium spp.(Biotechnology Letters, 2002-05) Pereira, Jorge Fernando; Queiroz, Marisa Vieira de; Gomes, Eliane Aparecida; Muro-Abad, Júpiter Israel; Araújo, Elza Fernandes dePenicillium species were analyzed with molecular markers and for pectinase and cellulase production. RAPD and PCR-RFLP analysis indicated high polymorphism among at least 5 of 10 Penicillium species. Five species were chosen for pectinase and cellulase production in liquid medium and four of which appeared similar based on molecular analyses. P. brevicompactum and P. griseoroseum gave the highest pectinase production and were highly divergent by molecular techniques.Item Avaliação da diversidade genética de fungos micorrízicos arbusculares em três estádios de estabilização de dunas(Revista Brasileira de Ciência do Solo, 2002-06) Araújo, E. F.; Mendonça, M. M.; Córdoba, A. S.A diversidade genética de fungos micorrízicos arbusculares, recuperados de três estádios de estabilização de dunas, foi avaliada por técnicas moleculares e comparada com resultados obtidos anteriormente por técnicas baseadas na caracterização morfológica dos esporos. O uso da técnica de PCR-RFLP do rDNA, extraído de esporos, permitiu definir impressões características de espécies presentes nas dunas, evidenciar a presença de diferentes comunidades em cada estádio e identificar a anteduna como aquela com comunidades com maior polimorfismo. Esse estádio também apresentou maior diversidade, quando, no estudo das comunidades, foram utilizadas técnicas baseadas em aspectos morfológicos. A combinação de ambas as estratégias, molecular e baseada em aspectos morfológicos, forneceu importantes informações sobre a diversidade destes fungos, visando ao estudo do seu papel no ecossistema.Item Differential expression of polygalacturonase-encoding genes from Penicillium griseoroseum in different carbon sources(Journal of Industrial Microbiology and Biotechnology, 2002-06-14) Ribon, A. O. B.; Queiroz, M. V.; Coelho, J. L. C.; Araújo, E. F deA second polygalacturonase - encoding gene ( pgg2 ) of Penicillium griseoroseum was cloned and consists of an opening reading frame of 1107 bp after removal of two introns. The gene encodes a protein of 369 amino acids with a predicted molecular mass of 38.3 kDa. The deduced protein sequence exhibited high homology with other fungal endopolygalacturonases. A polymerase chain reaction ( PCR ) - based strategy was used to study the expression patterns of pgg1 and pgg2 genes under different culture conditions and our results show that both genes are regulated by the carbon source at the transcriptional level. The pgg1 transcript was detected at 76 h of fungal growth in pectin while the pgg2 transcript was also induced by sucrose. The addition of yeast extract to glucose medium abolished the repressive effect of glucose, suggesting that the transcription of these genes is controlled by different mechanisms.Item Influência de doses de calcário com diferentes relações cálcio: magnésio na produção de matéria seca e na composição mineral da alfafa(Pesquisa Agropecuária Brasileira, 2002-12) Gomes, Fernando Teixeira; Borges, Arnaldo Chaer; Neves, Júlio César Lima; Fontes, Paulo Cezar RezendeA acidez dos solos brasileiros, com alumínio em níveis tóxicos e baixa disponibilidade de elementos essenciais, principalmente o fósforo, exige aplicações de calcário e fertilizantes para a adequada utilização agrícola. Este trabalho objetivou avaliar o efeito de quatro doses de calcário calculadas com base no teor de Al3+ trocável do solo e em cinco relações molares de Ca:Mg, na produção de matéria seca e na composição mineral da alfafa. Duas cultivares de alfafa, Flórida 77 e Crioula, foram cultivadas em Latossolo Vermelho-Amarelo álico, textura argilosa, em casa de vegetação, em esquema fatorial disposto no delineamento inteiramente casualizado, com quatro repetições. A produção de matéria seca da parte aérea das duas cultivares aumentou em razão do aumento das doses de calcário, e somente a cultivar Flórida 77 apresentou diferenças significativas entre as relações molares Ca:Mg. Os teores de Ca, Mg e N na matéria seca da parte aérea, de modo geral, aumentaram em razão do aumento na quantidade de calcário, sendo somente os teores de Ca e Mg alterados pelas diferentes relações molares de Ca:Mg. Os teores de P e K, de modo geral, apresentaram pequenos decréscimos com a elevação das doses de calcário, embora considerados satisfatórios para a nutrição da alfafa.