Endophytic fungi from Vitis labrusca L. ('Niagara Rosada') and its potential for the biological control of Fusarium oxysporum

We investigated the diversity of endophytic fungi found on grape (Vitis labrusca cv. Niagara Rosada) leaves collected from Salesopolis, SP, Brazil. The fungi were isolated and characterized by amplified ribosomal DNA restriction analysis, followed by sequencing of the ITS1-5.8S-ITS2 rDNA. In addition, the ability of these endophytic fungi to inhibit the grapevine pathogen Fusarium oxysporum f. sp herbemontis was determined in vitro. We also observed that the climatic factors, such as temperature and rainfall, have no effect on the frequency of infection by endophytic fungi. The endophytic fungal community that was identified included Aporospora terricola, Aureobasidium pullulans, Bjerkandera adusta, Colletotrichum boninense, C. gloeosporioides, Diaporthe helianthi, D. phaseolorum, Epicoccum nigrum, Flavodon flavus, Fusarium subglutinans, F. sacchari, Guignardia mangiferae, Lenzites elegans, Paraphaeosphaeria pilleata, Phanerochaete sordida, Phyllosticta sp, Pleurotus nebrodensis, Preussia africana, Tinctoporellus epiniltinus, and Xylaria berteri. Among these isolates, two, C. gloeosporioides and F. flavus, showed potential antagonistic activity against F. oxysporum f. sp herbemontis. We suggest the involvement of the fungal endophyte community of V. labrusca in protecting the host plant against pathogenic Fusarium species. Possibly, some endophytic isolates could be selected for the development of biological control agents for grape fungal disease; alternatively, management strategies could be tailored to increase these beneficial fungi.State of Sao Paulo Research Foundation (FAPESP/BIOTA)State of Sao Paulo Research Foundation (FAPESP/BIOTA) [2004/13910-6]National Council for Scientific and Technological Development (CNPq)National Council for Scientific and Technological Development (CNPq

CONFLITOS DA ARBORIZAÇÃO COM ELEMENTOS URBANOS NA CIDADE DE VALENÇA, ESTADO DO RIO DE JANEIRO

O planejamento da arborização tem grande importância na gestão urbana, a fim de evitar ou minimizar os conflitos que ainda podem causar prejuízos nas estruturas públicas ou privadas. Nesse estudo foi realizado um inventário da arborização na região central de Valença, cidade turística do estado do Rio de Janeiro, com registro dos conflitos de árvores com a fiação, acesso, iluminação, portões e garagens, recuo do meio fio e serviços subterrâneos. As ruas foram categorizadas e a maior parte delas apresentou característica de via residencial (67,3%) seguida de uso misto (20,8%) e por fim uso comercial (11,9%). Do total de árvores levantadas, 45% apresentam conflitos com a fiação telefônica e 38% com a fiação elétrica, os maiores valores dentre todos os elementos urbanos. Um número bastante alto apresentou sinais de podas, indicando conflitos anteriores e demonstrando forte intervenção nestes indivíduos. Estes resultados estão de acordo com estudos realizados em diferentes espaços urbanos onde não foi observado planejamento, que também mostram percentuais elevados de conflitos e uma alta demanda por intervenções

Analysis of 16S rRNA and mxaF genes reveling insights into Methylobacterium niche-specific plant association

The genus Methylobacterium comprises pink-pigmented facultative methylotrophic (PPFM) bacteria, known to be an important plant-associated bacterial group. Species of this group, described as plant-nodulating, have the dual capacity of producing cytokinin and enzymes, such as pectinase and cellulase, involved in systemic resistance induction and nitrogen fixation under specific plant environmental conditions. The aim hereby was to evaluate the phylogenetic distribution of Methylobacterium spp. isolates from different host plants. Thus, a comparative analysis between sequences from structural (16S rRNA) and functional mxaF (which codifies for a subunit of the enzyme methanol dehydrogenase) ubiquitous genes, was undertaken. Notably, some Methylobacterium spp. isolates are generalists through colonizing more than one host plant, whereas others are exclusively found in certain specific plant-species. Congruency between phylogeny and specific host inhabitance was higher in the mxaF gene than in the 16S rRNA, a possible indication of function-based selection in this niche. Therefore, in a first stage, plant colonization by Methylobacterium spp. could represent generalist behavior, possibly related to microbial competition and adaptation to a plant environment. Otherwise, niche-specific colonization is apparently impelled by the host plant

Environmental interactions are regulated by temperature in Burkholderia seminalis TC3.4.2R3.

Burkholderia seminalis strain TC3.4.2R3 is an endophytic bacterium isolated from sugarcane roots that produces antimicrobial compounds, facilitating its ability to act as a biocontrol agent against phytopathogenic bacteria. In this study, we investigated the thermoregulation of B. seminalis TC3.4.2R3 at 28 °C (environmental stimulus) and 37 °C (host-associated stimulus) at the transcriptional and phenotypic levels. The production of biofilms and exopolysaccharides such as capsular polysaccharides and the biocontrol of phytopathogenic fungi were enhanced at 28 °C. At 37 °C, several metabolic pathways were activated, particularly those implicated in energy production, stress responses and the biosynthesis of transporters. Motility, growth and virulence in the Galleria mellonella larvae infection model were more significant at 37 °C. Our data suggest that the regulation of capsule expression could be important in virulence against G. mellonella larvae at 37 °C. In contrast, B. seminalis TC3.4.2R3 failed to cause death in infected BALB/c mice, even at an infective dose of 107 CFU.mL-1. We conclude that temperature drives the regulation of gene expression in B. seminalis during its interactions with the environment

Isolation, identification and genetic characterization of endophytic bacteria from rootstocks citrus

Endófitos são microrganismos que em pelo menos uma fase do seu ciclo de vida, habitam o interior de tecidos da parte aérea de plantas assintomáticas. Foi realizado um estudo sobre a população bacteriana endofítica de dez porta-enxertos de citros, mantidos no Centro de Citricultura Sylvio Moreira - IAC. Foram isolados de explantes foliares principalmente dois grupos bacterianos: um deles constituído de isolados do gênero Bacillus (incluindo B. pumilus, B. megaterium, B. cereus, B. lentus) e outro, constituído por Erwinia herbicola. Verificou-se que a espécie mais frequente foi E. herbicola (24,45%); entretanto, se for considerado o gênero, Bacillus foi o mais frequente (50,98%). A presença em maior frequência de isolados deste gênero se deve provavelmente à resistência de seus endósporos ao tratamento para desinfecção superficial dos tecidos foliares, pois resultados obtidos da interação destas bactérias com o fungo endofítico Guignardia citricarpa e análise via marcadores de RAPD, sugerem que Bacillus spp. fazem parte da população epifítica destas plantas, e eventualmente podem entrar no hospedeiro por meio de aberturas como estômatos elou ferimentos, sendo nestes casos considerados como endófitos facultativos. Já, E. herbicola apresenta uma associação simbiótica com o fungo endofítico estudado, o qual deve favorecer a colonização bacteriana no hospedeiro. Esta característica aliada ao fato de que esta bactéria, frequentemente isolada de plantas como endófita, apresentou pequena divergência genética, detectada por marcadores de RAPD e perfil plasmidial, sugerem que deva ocorrer uma íntima interação entre o fungo G. citricarpa, a bactéria endofítica e a planta hospedeira, a qual deve manter reduzida a variabilidade genética neste grupo bacteriano. A constatação do estado endofítico de E. herbicola, juntamente com esta interação com a planta, mostra a possibilidade de se utilizar esta espécie para a introdução de características de interesse em porta-enxertos de citros.Endophytes are microorganisms which live inside aerial tissue of assyntomatic plants, during at least one phase of their life cycles. Ten rootstocks citrus were used. In order to get insights about their endophytic bacterial population. Endophytic bacteria were isolated from foliar tissue and could be divided in two main groups. The first group was formed by Bacillus (including B. megaterium, B. cereus, B. pumilus, B. lentus), and a second group was formed by Erwinia herbicola. The frequencies of bacteria from group one and two were 50,98% and 24,45%, respectively. The high frequency of the genus Bacillus found in the bacterial population is probably due to the endospore resistance to superficial desinfection of the leaf tissue. This observation was based on the interaction showed by these bacterias with the endophytic fungi Guignardia citricarpa and RAPD analysis, suggesting that Bacillus spp. is part of the epiphytic population of this plants. Eventually these epiphytic bacteria it may penetrate inside the host by stomats or injuries and in this case they could be considered facultative endophytes. However, E herbicola shows a simbiotic association with the studied endophytic fungi, which favours the bacterial colonization of the host. This finding, plus the low genetic divergence of several isolates from this bacteria, detected by RAPD and plasmidial profile, suggested that E. herbicola occurs in a close interaction with the fungus. The endophytic state of E. herbicola, added to its interaction with the host plants, suggests the possibility of using this species for the introduction of characteristics of interest in rootstocks citrus

Isolation, identification and genetic characterization of endophytic bacteria from rootstocks citrus

Endófitos são microrganismos que em pelo menos uma fase do seu ciclo de vida, habitam o interior de tecidos da parte aérea de plantas assintomáticas. Foi realizado um estudo sobre a população bacteriana endofítica de dez porta-enxertos de citros, mantidos no Centro de Citricultura Sylvio Moreira - IAC. Foram isolados de explantes foliares principalmente dois grupos bacterianos: um deles constituído de isolados do gênero Bacillus (incluindo B. pumilus, B. megaterium, B. cereus, B. lentus) e outro, constituído por Erwinia herbicola. Verificou-se que a espécie mais frequente foi E. herbicola (24,45%); entretanto, se for considerado o gênero, Bacillus foi o mais frequente (50,98%). A presença em maior frequência de isolados deste gênero se deve provavelmente à resistência de seus endósporos ao tratamento para desinfecção superficial dos tecidos foliares, pois resultados obtidos da interação destas bactérias com o fungo endofítico Guignardia citricarpa e análise via marcadores de RAPD, sugerem que Bacillus spp. fazem parte da população epifítica destas plantas, e eventualmente podem entrar no hospedeiro por meio de aberturas como estômatos elou ferimentos, sendo nestes casos considerados como endófitos facultativos. Já, E. herbicola apresenta uma associação simbiótica com o fungo endofítico estudado, o qual deve favorecer a colonização bacteriana no hospedeiro. Esta característica aliada ao fato de que esta bactéria, frequentemente isolada de plantas como endófita, apresentou pequena divergência genética, detectada por marcadores de RAPD e perfil plasmidial, sugerem que deva ocorrer uma íntima interação entre o fungo G. citricarpa, a bactéria endofítica e a planta hospedeira, a qual deve manter reduzida a variabilidade genética neste grupo bacteriano. A constatação do estado endofítico de E. herbicola, juntamente com esta interação com a planta, mostra a possibilidade de se utilizar esta espécie para a introdução de características de interesse em porta-enxertos de citros.Endophytes are microorganisms which live inside aerial tissue of assyntomatic plants, during at least one phase of their life cycles. Ten rootstocks citrus were used. In order to get insights about their endophytic bacterial population. Endophytic bacteria were isolated from foliar tissue and could be divided in two main groups. The first group was formed by Bacillus (including B. megaterium, B. cereus, B. pumilus, B. lentus), and a second group was formed by Erwinia herbicola. The frequencies of bacteria from group one and two were 50,98% and 24,45%, respectively. The high frequency of the genus Bacillus found in the bacterial population is probably due to the endospore resistance to superficial desinfection of the leaf tissue. This observation was based on the interaction showed by these bacterias with the endophytic fungi Guignardia citricarpa and RAPD analysis, suggesting that Bacillus spp. is part of the epiphytic population of this plants. Eventually these epiphytic bacteria it may penetrate inside the host by stomats or injuries and in this case they could be considered facultative endophytes. However, E herbicola shows a simbiotic association with the studied endophytic fungi, which favours the bacterial colonization of the host. This finding, plus the low genetic divergence of several isolates from this bacteria, detected by RAPD and plasmidial profile, suggested that E. herbicola occurs in a close interaction with the fungus. The endophytic state of E. herbicola, added to its interaction with the host plants, suggests the possibility of using this species for the introduction of characteristics of interest in rootstocks citrus

Epicolactone - Natural Product Isolated from the Sugarcane Endophytic Fungus Epicoccum nigrum

The endophytic fungus Epicoccum nigrum was isolated from sugarcane and the bioguided fractionation of the ethyl acetate extract led to the isolation of epicolactone, mellein, and 4,5-dimethylresorcinol. Characterization of epicolactone by MS, NMR and X-ray crystallography revealed a new natural product with an unusual carbon skeleton. The production of this secondary metabolite decreased in mutants of Epicoccum nigrum transformed by Agrobacterium tumefaciens. Additionally, these mutants produced 4-hydroxymellein.Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [2010/08286-2]Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)PetrobrasPETROBRA

Epicolactone - Natural Product Isolated from the Sugarcane Endophytic Fungus Epicoccum nigrum

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)The endophytic fungus Epicoccum nigrum was isolated from sugarcane and the bioguided fractionation of the ethyl acetate extract led to the isolation of epicolactone, mellein, and 4,5-dimethylresorcinol. Characterization of epicolactone by MS, NMR and X-ray crystallography revealed a new natural product with an unusual carbon skeleton. The production of this secondary metabolite decreased in mutants of Epicoccum nigrum transformed by Agrobacterium tumefaciens. Additionally, these mutants produced 4-hydroxymellein.2752255230Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)PetrobrasFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Participação em colónias de férias e promoção da autonomia : perceções de adolescentes e monitores

A autonomia traduz-se numa habilidade relacional cujo desenvolvimento ocorre ao longo de toda a vida sendo influenciada por variáveis individuais, contextuais e familiares diversas (Barbosa & Wagner, 2015). O estudo apresentado procura compreender a autonomia, percecionada por 42 adolescentes/jovens e por 66 monitores/animadores, e o contributo da participação em Colónias de Férias neste âmbito, atendendo a variáveis sociodemográficas e institucionais. A amostra de adolescentes/jovens foi sobretudo do litoral (69%), género feminino (76.2%) e 68.8% que ainda se encontravam institucionalizados. Quanto aos monitores, a maioria era do litoral (77.3%), encontrava-se a trabalhar (84.8%), 63.6% do género masculino e apresentava idade média de 28 anos. Foi aplicado aos adolescentes o Questionário de Autonomia dos Adolescentes e um instrumento construído especificamente para o estudo aos monitores. Recorreu-se ao SPSS IBM 23 para a análise de dados. Relativamente à autonomia dos adolescentes/jovens percecionada pelos próprios verificaram-se diferenças estatisticamente significativas entre: 1) rapazes (melhores resultados) e raparigas na independência e autonomia cognitiva; 2) grupos etários (13-17 vs. 18-24) na auto-determinação (resultados superiores dos 13 aos 17); 3) os que residiam ainda em instituição ou não (médias mais elevadas) na independência; 4) e adolescentes e jovens institucionalizados (resultados mais favoráveis) e não institucionalizados na auto-determinação e autonomia total. Em relação à contribuição das Colónias de Férias para a autonomia dos adolescentes e jovens, percecionada pelos próprios e pelos monitores/animadores, não se verificaram resultados estatisticamente significativos. Será fundamental continuar a explorar estas questões inerentes à compreensão da autonomia, nomeadamente atendendo a variáveis de natureza diversa, incluindo aspetos pessoais e contextuais.The autonomy refers to a relational hability that grows lifelong influenced by individual, contextual and familiar variables (Barbosa & Wagner, 2015). The following study aims to understand the autonomy perceived by 42 teenagers/young people and 66 monitors/animators as well as the contribution of participations in Holidays Camps in this sense, considering sociodemografic and institucional. The teenagers/youngs were in large number from coast (69%), females (76.2%) and people who still are institutionalized (68.8%). As for the monitors, the majority lived in the coast (77.3%), most of them had a job (84.8%) and there were more males (63.6%), with the medium age of 28 years. The Adolescents Autonomy Questionnaire was applied to adolescents and an instrument built specifically for the study of the monitors. SPSS IBM23 was used for data analysis. For the teenagers and young's autonomy, there was some statistically significant differences between: 1- boys (better results) and girls on independence and cognitive autonomy; 2- age groups (13-17 vs 18-24) on self-determination (higher levels on 13-17 group); 3- those who still live in institution and those who don’t (higher results), on independence; 4- institutionalized and non-institutionalized individuals (better results) on self-determination and total autonomy. In relation to the contribution of Holidays Camps for the teenagers and youngs autonomy, there were not statistically significant results. Its fundamental that we continue to explore this questions about autonomy to have a better knowledge, particularly about the contribution of the personal and contextual variables

Complete genome sequence of the sugarcane nitrogen-fixing endophyte gluconacetobacter diazotrophicus PAL5

17 p. : il.Background: Gluconacetobacter diazotrophicus Pal5 is an endophytic diazotrophic bacterium that lives in association with sugarcane plants. It has important biotechnological features such as nitrogen fixation, plant growth promotion, sugar metabolism pathways, secretion of organic acids, synthesis of auxin and the occurrence of bacteriocins. Results: Gluconacetobacter diazotrophicus Pal5 is the third diazotrophic endophytic bacterium to be completely sequenced. Its genome is composed of a 3.9 Mb chromosome and 2 plasmids of 16.6 and 38.8 kb, respectively. We annotated 3,938 coding sequences which reveal several characteristics related to the endophytic lifestyle such as nitrogen fixation, plant growth promotion, sugar metabolism, transport systems, synthesis of auxin and the occurrence of bacteriocins. Genomic analysis identified a core component of 894 genes shared with phylogenetically related bacteria. Gene clusters for gum-like polysaccharide biosynthesis, tad pilus, quorum sensing, for modulation of plant growth by indole acetic acid and mechanisms involved in tolerance to acidic conditions were identified and may be related to the sugarcane endophytic and plant-growth promoting traits of G. diazotrophicus. An accessory component of at least 851 genes distributed in genome islands was identified, and was most likely acquired by horizontal gene transfer. This portion of the genome has likely contributed to adaptation to the plant habitat. Conclusion: The genome data offer an important resource of information that can be used to manipulate plant/bacterium interactions with the aim of improving sugarcane crop production and other biotechnological applications