Oxidative foliar photo-necrosis produced by the bacteria Pseudomonas cedrina

BACKGROUND: Although bioactive metabolites capable of causing oxidative photo-necrosis in plant tissues have been identified in fungi, little is known about this type of mechanism in bacteria. These metabolites act as photosensitizers that generate reactive oxygen species (ROS) capable of causing damage to cells. In addition, these metabolites can pass into an energetically excited state when they receive some luminous stimulus, a condition in which they interact with other molecules present in the environment, such as molecular oxygen (O2), also known as triplet oxygen (3 O2), generating ROS. RESULTS: The suspension of the bacterial culture of Pseudomonas cedrina was shown to produce foliar necrosis in papaya leaves (Carica papaya L.) only in the presence of sunlight, which is evidence of photosensitizing mechanisms that generate singlet oxygen (1 O2). From the chemical study of extracts obtained from this bacteria, 3-(4-(2-carboxipropyl) phenyl) but-2-enoic acid (1) was isolated. This compound, in the presence of light and triplet oxygen (3 O2), was able to oxidize ergosterol to its peroxide, since it acted as a photosensitizer producing 1 O2, with which it was corroborated that a photosensitization reaction occurs, mechanism by which this bacterium could prove to cause oxidative foliar photo-necrosis. CONCLUSIONS: P. cedrina was able to induce oxidative foliar photo-necrosis because of its potential ability to produce photosensitizing metabolites that generate singlet oxygen in the plants it colonizes. Based on the above, it can be proposed that some bacteria can cause oxidative foliar photo-necrosis as an important mechanism in the pathogenesis of host species.

Illumina-based analysis of endophytic bacterial diversity of tree peony (Paeonia Sect. Moutan) roots and leaves

ABSTRACT Diverse communities of bacteria inhabit plant tissues and those bacteria play a crucial role for plant health and growth. Tree peony (Paeonia Sect. Moutan) is known for its excellent ornamental and medicinal values as Chinese traditional plant, but little is known about its associated bacterial community under natural conditions. To examine how endophytic bacteria in tree peony vary across tissues and cultivars, PCR-based Illumina was applied to reveal the diversity of endophytic bacteria in tree peony. A total of 149,842 sequences and 21,463 operational taxonomic units (OTUs) were obtained. The OTU abundance of roots was higher than leaves across other three cultivars except for 'Kinkaku' and 'Luoyanghong'. The community was composed of five dominant groups (Proteobacteria, Firmicutes, Bacteroidetes, Acidobacteria and Actinobacteria) in all samples. Endophytic bacteria community structures had changed in leaves and roots. Sequences of Pseudomonas and Enterobacteriaceae were prevalent in root samples, whereas Succinivibrio and Acinetobacter were the dominant genus in leaf samples. Otherwise, the distribution of each dominant genus among the 5 cultivars was either varied. These findings suggested that both plant genotype and tissues contribute to the shaping of the bacterial communities associated with tree peony.

Endophytic fungi from selected varieties of soybean (Glycine max L. Merr.) and corn (Zea mays L.) grown in an agricultural area of Argentina / Hongos endófitos aislados de cultivares de soja (Glycine max L. Merr) y maíz (Zea mays L.) presentes en áreas agrícolas argentinas

Endophytic fungi are ubiquitous and live within host plants without causing any noticeable symptoms of disease. Little is known about the diversity and function of fungal endophytes in plants, particularly in economically important species. The aim of this study was to determine the identity and diversity of endophytic fungi in leaves, stems and roots of soybean and corn plants and to determine their infection frequencies. Plants were collected in six areas of the provinces of Buenos Aires and Entre Ríos (Argentina) two areas were selected for sampling corn and four for soybean. Leaf, stem and root samples were surface-sterilized, cut into 1 cm² pieces using a sterile scalpel and aseptically transferred to plates containing potato dextrose agar plus antibiotics. The species were identified using both morphological and molecular data. Fungal endophyte colonization in soybean plants was influenced by tissue type and varieties whereas in corn plants only by tissue type. A greater number of endophytes were isolated from stem tissues than from leaves and root tissues in both species of plants. The most frequently isolated species in all soybean cultivars was Fusarium graminearum and the least isolated one was Scopulariopsis brevicaulis. Furthermore, the most frequently isolated species in corn plants was Aspergillus terreus whereas the least isolated one was Aspergillus flavus. These results could be relevant in the search for endophytic fungi isolates that could be of interest in the control of agricultural pests.

Los hongos endófitos son ubicuos y se encuentran en el interior de los tejidos de las plantas de manera asintomática. Se sabe muy poco acerca de la diversidad y la función de estos hongos, particularmente en especies de importancia económica. El objetivo de este trabajo fue determinar la diversidad y la frecuencia de colonización de hongos endófitos en raíces, tallos y hojas de 2 variedades de maíz y de 4 variedades de soja; las muestras se tomaron de 6 áreas diferentes ubicadas en las provincias de Buenos Aires y Entre Ríos (Argentina). Con un bisturí estéril se obtuvieron porciones de 1 cm² de raíz, tallo y hoja, que fueron colocados en placas con agar papa dextrosa más antibiótico. Las especies de hongos fueron identificadas a partir de características morfológicas y moleculares. La colonización de hongos endófitos en soja estuvo influenciada por la variedad y por el tipo de tejido, en tanto que en el maíz solo hubo influencia del tipo de tejido. El mayor número de endófitos se encontró en los tallos de ambas especies. El aislamiento más frecuente en todas las variedades de soja fue Fusarium graminearum y el menos frecuente Scopulariopsis brevicaulis. En ambas variedades de maíz la especie con mayor frecuencia de aislamiento fue Aspergillus terreus y la de menor fue Aspergillus flavus. Estos resultados son relevantes para la búsqueda de especies de hongos endófitos que podrían ser de interés en el control de plagas agrícolas.

Characterization saprobic fungi on leaf litter of two species of trees in the Atlantic Forest, Brazil

Abstract We investigated the composition and structure of fungal communities associated with leaf litter generated by Clusia nemorosa and Vismia guianensis that belong to phylogenetically-related botanical families and exist together in a remnant of the Atlantic Forest in Bahia, Brazil. Samplings were conducted during wet (June 2011) and dry (January 2013) seasons in Serra da Jibóia. The fungi were isolated using particle filtration and the 1,832 isolates represented 92 taxa. The wet season yielded the largest number of isolates (1,141) and taxa (76) compared with the dry season (641 isolates and 37 taxa). The richness and diversity of fungal species associated with C. nemorosa (64 taxa, Simpson=0.95)were higher compared with those of V.guianensis (59 taxa, Simpson =0.90). Analysis of similarity (ANOSIM) revealed significant variations in the composition and community structure of fungi isolated from the two plants as a function of seasons. In contrast, nonmetric multidimensional scaling (NMDS) analysis show that the seasonality was an important influence on the distribution of fungal species. However, the populations of the saprobic fungal communities were dynamic, and several factors may influence such communities in the Atlantic Forest.

Characterization of the hrpZ gene from Pseudomonas syringae pv. maculicolaM2

Pseudomonas syringae pv. maculicola is a natural pathogen of members of the Brassicaceae plant family. Using a transposon-based mutagenesis strategy in Pseudomonas syringaepv. maculicola M2 (PsmM2), we conducted a genetic screen to identify mutants that were capable of growing in M9 medium supplemented with a crude extract from the leaves of Arabidopsis thaliana. A mutant containing a transposon insertion in the hrpZ gene (PsmMut8) was unable to infect adult plants from Arabidopsis thaliana or Brassica oleracea, suggesting a loss of pathogenicity. The promotorless cat reporter present in the gene trap was expressed if PsmMut8 was grown in minimal medium (M9) supplemented with the leaf extract but not if grown in normal rich medium (KB). We conducted phylogenetic analysis using hrpAZB genes, showing the classical 5-clade distribution, and nucleotide diversity analysis, showing the putative position for selective pressure in this operon. Our results indicate that the hrpAZB operon from Pseudomonas syringaepv. maculicola M2 is necessary for its pathogenicity and that its diversity would be under host-mediated diversifying selection.

Characterization of the hrpZ gene from Pseudomonas syringae pv. maculicolaM2

Pseudomonas syringae pv. maculicola is a natural pathogen of members of the Brassicaceae plant family. Using a transposon-based mutagenesis strategy in Pseudomonas syringaepv. maculicola M2 (PsmM2), we conducted a genetic screen to identify mutants that were capable of growing in M9 medium supplemented with a crude extract from the leaves of Arabidopsis thaliana. A mutant containing a transposon insertion in the hrpZ gene (PsmMut8) was unable to infect adult plants from Arabidopsis thaliana or Brassica oleracea, suggesting a loss of pathogenicity. The promotorless cat reporter present in the gene trap was expressed if PsmMut8 was grown in minimal medium (M9) supplemented with the leaf extract but not if grown in normal rich medium (KB). We conducted phylogenetic analysis using hrpAZB genes, showing the classical 5-clade distribution, and nucleotide diversity analysis, showing the putative position for selective pressure in this operon. Our results indicate that the hrpAZB operon from Pseudomonas syringaepv. maculicola M2 is necessary for its pathogenicity and that its diversity would be under host-mediated diversifying selection.

Cultivable endophytic bacteria from leaf bases of Agave tequilana and their role as plant growth promoters

Agave tequilana Weber var. 'Azul' is grown for the production of tequila, inulin and syrup. Diverse bacteria inhabit plant tissues and play a crucial role for plant health and growth. In this study culturable endophytic bacteria were extracted from leaf bases of 100 healthy Agave tequilana plants. In plant tissue bacteria occurred at mean population densities of 3 million CFU/g of fresh plant tissue. Three hundred endophytic strains were isolated and 16s rDNA sequences grouped the bacteria into eight different taxa that shared high homology with other known sequences. Bacterial endophytes were identified as Acinectobacter sp., A. baumanii, A. bereziniae, Cronobacter sakazakii, Enterobacter hormaechei, Bacillus sp. Klebsiella oxytoca, Pseudomonas sp., Enterococcus casseliflavus, Leuconostoc mesenteroides subsp. mesenteroides and Gluconobacter oxydans. Isolates were confirmed to be plant growth promoting bacteria (PGPB) by their capacities for nitrogen fixation, auxin production, phosphate solubilization, or antagonism against Fusarium oxysporum AC132. E. casseliflavus JM47 and K. oxytoca JM26 secreted the highest concentrations of IAA. The endophyte Acinectobacter sp. JM58 exhibited the maximum values for nitrogen fixation and phosphate solubilization index (PSI). Inhibition of fungi was found in Pseudomonas sp. JM9p and K. oxytoca JM26. Bacterial endophytes show promise for use as bio-inoculants for agave cultivation. Use of endophytes to enhance cultivation of agave may be particularly important for plants produced by micropropagation techniques, where native endophytes may have been lost.

Production and characterization of an extracellular lipase from Candida guilliermondii

Extracellular lipases from the endophytic yeast Candida guilliermondii isolated from castor leaves (Ricinus communis L.) were produced using low-cost raw materials such as agro-industrial residues and applying them in the esterification of oleic acid for evaluating their potential use in biodiesel production. After partial purification using ammonium sulfate, the enzyme was characterized and presented higher activity (26.8 ± 1.5 U mL-1) in the presence of 5 mmol L-1 NaCl at 30 ºC and pH 6.5. The production through submerged fermentation was formerly performed in 150 mL erlenmeyer flasks and, once the enzyme production was verified, assays in a 14 L bioreactor were conducted, obtaining 18 ± 1.4 U mL-1. The produced enzyme was applied in the oleic acid esterification under different solvents: hexane, cyclohexane or cyclohexanone) and different acid:alcohol molar ratios. Higher ester conversion rate (81%) was obtained using hexane and the molar ratio of 1:9 was the best conditions using methanol. The results suggest the potential for development of endophytic yeast in the production of biocatalyst through submerged fermentation using agroindustrial residues as culture medium.

A procedure to evaluate the efficiency of surface sterilization methods in culture-independent fungal endophyte studies

Extraneous DNA interferes with PCR studies of endophytic fungi. A procedure was developed with which to evaluate the removal of extraneous DNA. Wheat (Triticum aestivum) leaves were sprayed with Saccharomyces cerevisiae and then subjected to physical and chemical surface treatments. The fungal ITS1 products were amplified from whole tissue DNA extractions. ANOVA was performed on the DNA bands representing S. cerevisiae on the agarose gel. Band profile comparisons using permutational multivariate ANOVA (PERMANOVA) and non-metric multidimensional scaling (NMDS) were performed on DGGE gel data, and band numbers were compared between treatments. Leaf surfaces were viewed under variable pressure scanning electron microscopy (VPSEM). Yeast band analysis of the agarose gel showed that there was no significant difference in the mean band DNA quantity after physical and chemical treatments, but they both differed significantly (p < 0.05) from the untreated control. PERMANOVA revealed a significant difference between all treatments (p < 0.05). The mean similarity matrix showed that the physical treatment results were more reproducible than those from the chemical treatment results. The NMDS showed that the physical treatment was the most consistent. VPSEM indicated that the physical treatment was the most effective treatment to remove surface microbes and debris. The use of molecular and microscopy methods for the post-treatment detection of yeast inoculated onto wheat leaf surfaces demonstrated the effectiveness of the surface treatment employed, and this can assist researchers in optimizing their surface sterilization techniques in DNA-based fungal endophyte studies.

Endophytic fungi from Myrcia guianensis at the Brazilian Amazon: distribution and bioactivity

Beneficial interactions between plants and microorganisms have been investigated under different ecological, physiological, biochemical, and genetic aspects. However, the systematic exploration of biomolecules with potential for biotechnological products from this interaction still is relatively scarce. Therefore, this study aimed the evaluation of the diversity and antimicrobial activity of the endophytic fungi obtained from roots, stems and leafs of Myrcia guianensis (Myrtaceae) from the Brazilian Amazon. 156 endophytic fungi were isolated and above 80% were identified by morphological examination as belonging to the genera Pestalotiopsis, Phomopsis, Aspergillus, Xylaria, Nectria, Penicillium and Fusarium. Fermented broth of those fungi were assayed for antimicrobial activity and four inhibited the growth of Staphylococcus aureus, Enterococcus faecalis, Candida albicans and Penicillium avellaneum. As the strain named MgRe2.2.3B (Nectria haematococca) had shown the most promising results against those pathogenic strains, its fermented broth was fractioned and only its two low polar fractions demonstrated to be active. Both fractions exhibited a minimum bactericidal concentration of 50 µg.mL-1 against S. aureus and a minimum fungicidal concentration of 100 µg.mL-1 against P. avellaneum. These results demonstrate the diversity of fungal genera in M. guianensis and the potential of these endophytic fungi for the production of new antibiotics.

Ultrastructural changes and oxidative stress markers in wild and cultivar Sesamum orientale L. following Alternaria sesami (Kawamura) Mohanty and Behera inoculation.

Alternaria sesami causes leaf spot disease in Sesamum orientale. Conidium germination, inoculation, penetration and colonization of the pathogen on the plant surfaces were studied using scanning electron microscopy. Electron microscopy analysis revealed multiple germ tubes from conidium that spread in all direction across the leaf surfaces. Penetration in the plant surface occured, directly through the epidermis or via stomata with or without the appressoria formation. Hyphal penetration continued through the substomata cavity and some of hyphal branches grew in the intercellular space of mesophyll tissue. Hyphal toxin, caused cell and cell wall damages. Changes in different biochemical parameters in the diseased sesame plants (both in wild and cultivar) were compared to control. Transmission electron microscopy showed structural changes in the chloroplast of diseased plants. Isozyme pattern and assays of different enzymes, namely catalase, acid phosphatase and peroxidase expressed varied level of activities. Meanwhile, esterase, polyphenol oxidase and superoxide dismutase in diseased plants showed remarkable levels compared to control. Due to the infection, chlorophyll content, carbohydrates and total soluble protein decreased whereas free amino acid, proline, phenols and disease-related proteins increased in the host plants. Differential SDS-PAGE band profiling of total soluble proteins were also observed in plants due to the infection.

Diversity of foliar endophytic fungi from the medicinal plant Sapindus saponaria L . and their localization by scanning electron microscopy

Endophytic fungi inhabit vegetable tissues or organs, without causing them any harm. Endophytes can co-evolve with plant hosts and possess species-specific interactions. They can protect the plant from insect attacks and diseases, and are also able to produce substances of biotechnological interest. In folk medicine, the bark, roots and fruits of Sapindus saponaria is used to produce substances with anxiolytic, astringent, diuretic and expectorant properties, as well as tonics, blood depuratives and cough medicine. This study evaluated the diversity of endophytic fungi present in the leaves of S. saponaria L. and observed the colonization of host plants by endophytes, using light and scanning electron microscopy. We verified that these fungi are found in intercellular and intracellular spaces. The genera of some isolates of S. saponaria were identified mainly by sequencing of ITS region of rDNA and, when possible, also by their microscopic features, as follows: Cochliobolus, Alternaria, Curvularia, Phomopsis, Diaporthe and Phoma. Phylogenetic analysis showed the existence of genetic variability of the genera Phomopsis and Diaporthe and interspecific variation among the Curvularia, Alternaria and Phoma, belonging to family Pleosporaceae.

Molecular battles between plant and pathogenic bacteria in the phyllosphere

The phyllosphere, i.e., the aerial parts of the plant, provides one of the most important niches for microbial colonization. This niche supports the survival and, often, proliferation of microbes such as fungi and bacteria with diverse lifestyles including epiphytes, saprophytes, and pathogens. Although most microbes may complete the life cycle on the leaf surface, pathogens must enter the leaf and multiply aggressively in the leaf interior. Natural surface openings, such as stomata, are important entry sites for bacteria. Stomata are known for their vital role in water transpiration and gas exchange between the plant and the environment that is essential for plant growth. Recent studies have shown that stomata can also play an active role in limiting bacterial invasion of both human and plant pathogenic bacteria as part of the plant innate immune system. As counter-defense, plant pathogens such as Pseudomonas syringae pv tomato (Pst) DC3000 use the virulence factor coronatine to suppress stomate-based defense. A novel and crucial early battleground in host-pathogen interaction in the phyllosphere has been discovered with broad implications in the study of bacterial pathogenesis, host immunity, and molecular ecology of bacterial diseases.

Danos causados pela infecção de oídio em diferentes estádios fenológicos da soja / Damages caused by infection of powdery mildews in different phenological stages of soybeans

Ainda não há estudos precisos que quantifiquem os prejuízos decorrentes de infecção por oídio e/ou outras doenças foliares, para a maioria das culturas de importância econômica no Brasil. O objetivo foi quantificar as perdas causadas por oídio (Microsphaera diffusa) infectando a cultura da soja em diferentes estádios fenológicos e relacioná-las ao desenvolvimento e produtividade da cultura. O experimento foi desenvolvido em ambiente protegido, e os tratamentos foram testemunha controlada, testemunha sem controle, infecção iniciada em R1 ­ R2, infecção iniciada em R ­ R, infecção iniciada em R ­ R e infecção iniciada em R ­ R. A avaliação foi feita 345.15.25.35.4 semanalmente, considerando a porcentagem da área foliar infectada. Os resultados mostraram que, no tratamento em que houve infecções iniciadas em R1-R2 e R3-R4, a porcentagem de área foliar afetada foi maior (41% e 38%, respectivamente), com consequente menor produtividade (1.186,6 e 1.309,5 kg.ha-1 respectivamente). No tratamento em que a infecção ocorreu em R ­ R, houve 5.35.4 a menor média de área foliar afetada pela doença (24%) e a produtividade teve queda de 26%. Os resultados mostraram que as perdas de produtividade pelo oídio na cultivar Embrapa 48 variaram ao redor de 26 a 50%, e que a recomendação oficial para o início de controle do oídio da soja, quando esta se apresentar entre 40 e 50% de severidade, deve ser questionada e outros trabalhos neste âmbito devem ser desenvolvidos para determinação das perdas ocasionadas por esta doença na cultura.

At present there are no precise studies quantifying the damages caused by powdery mildews and other foliar diseases for the majority of economically important crops in Brazil. The objective of the present study was to quantify the losses caused by powdery mildews (Microsphaera diffusa) in soybeans in different phenological stages, and to correlate them with the development and yield of the crop. The trials was carried out in the greenhouse and the treatments were: controlled check, noncontrolled check, infection initiated at stage R1-R2, infection initiated at stage R3-R4, infection initiated at stage R5.1-R5.2, infection initiated at stage R5.3-R5.4. The evaluation was done weekly considering the percentage of infected leaf area. The results showed that for the infection beginning at stages R1-R2 and R3-R4 the percentage of affected leaf area was higher (41% and 38%), with consequently lower yields (1,200 and 1,240 kg ha-1). When the infection occurred later at stage R5.3-R5.4, a lower affected leaf area (24%) was observed, and the yield decreased 26%. The results showed that the loss of yield by powdery mildew in cultivar Embrapa 48 ranged from around 26 to 50%, and that the official recommendation for the beginning of control of powdery mildew of soybean, where it presents between 40 and 50% of severity, should be questioned, and other work in this area should be undertaken to determine the loss caused by this disease in the crop.

Evaluation of foliar fungal endophyte diversity and colonization of medicinal plant Luehea divaricata (Martius et Zuccarini)

Endophyte microorganisms are organisms that live inside plants without causing any apparent damage to their hosts. Since all plants exhibit endophyte microorganisms, it is believed that mutual association is of great importance in nature. Luehea divaricata (Martius & Zuccarini), known popularly in Brazil as agoita-cavalo, is a big-sized tree with a wide distribution in the country that possesses medicinal qualities for: dysentery, leucorrhea, rheumatism, blennorrhoea, tumors, bronchitis, and depuration. This research aims at isolating and molecularly characterizing fungi isolates from L. divaricata by sequence analysis of ITS1-5.8S-ITS2 rDNA. Further, the colonization of endophyte in the host plant by Light and Scanning Electron Microscopy will also be investigated. Whereas, genera Alternaria, Cochliobolus, Diaporthe, Epicoccum, Guignardia, Phoma, and Phomopsis, were identified; rDNA sequence analysis revealed intra-species variability among endophyte isolates of the genus Phomopsis sp. Light and Scanning Electron Microscopy techniques showed the presence of endophyte fungi inside L. divaricata leaves, inhabiting inter- and intra-cellular spaces. These types of extensive colonization and dissemination were reported throughout all the leaf parts in palisade parenchyma, esclerenchyma, spongy parenchyma, adaxial epidermis, and vascular bundle indicating colonization of endophytes in múltiple structural sub-niches in the host plant.

Nutritional deficiency in citrus with symptoms of citrus variegated chlorosis disease / Deficiência nutricional em citros com sintomas da doença da clorose variegada dos citros

It is well known that citrus plants that have been infected by Xylella fastidiosa display nutritional deficiencies, probably caused by production of extracellular polymers by the bacteria that block normal nutrient flow through the xylem. The aim of this work was to study the mineral composition of specific foliar areas in different stages of infection in citrus. Thus, the concentrations of macro and micronutrients in leaves of citrus infected by X. fastidiosa were measured. Samples from four infected citrus orchards in the State of São Paulo, Brazil, were respectively collected from Santa Rita do Passa Quatro, Neves Paulista, Gavião Peixoto and Paraíso counties. The presence of X. fastidiosa in leaves was confirmed by polymerase chain reaction (PCR) using specific PCR primers. To understand the variation in leaf-nutrient content in citrus plants, we used foliar nutrient values from control (non-symptomatic) plants as a reference. Chemometric analysis showed that the deficiency of P and K in symptomatic trees for all orchards and high concentrations of Fe, Mn and Zn were observed in chlorotic areas, although other studies revealed deficiency of zinc in leaves. This is the first report showing that a correlation between chlorotic citrus leaf and higher concentrations of Fe, Mn and Zn are observed when infected and healthy plants were compared.

Já é bem conhecido que cultivares cítricas que foram infectadas pela bactéria Xylella fastidiosa apresentam deficiências nutricionais devido à produção de polímero extracelular por esta bactéria, o qual bloqueia o fluxo normal de nutriente pelo xilema. O objetivo deste trabalho foi o de estudar a composição mineral em áreas foliares específicas em diferentes fases de infecção na planta. Assim, as concentrações de macro e micronutrientes em folhas de citros infectados por X. fastidiosa foram quantificadas. Foram coletadas amostras de quatro pomares cítricos infectados localizados em: Santa Rita do Passa Quatro, Neves Paulista, Gavião Peixoto e Paraíso, no Estado de São Paulo. A presença de X. fastidiosa em folhas foi confirmada através de reação da polimerase em cadeia (PCR) usando iniciadores específicos. Para entender a variação no conteúdo de nutriente foliar em plantas cítricas, utilizou-se de valores de nutrientes foliares de plantas não sintomáticas (controle) como referência. A análise quimiométrica mostrou que a deficiência de P e K em plantas sintomáticas e concentrações altas de Fe, Mn e Zn foram presentes em áreas foliares cloróticas, embora outros estudos mostrem a deficiência de zinco em folhas. Este é o primeiro relato indicando que uma correlação entre folhas cítricas cloróticas e elevadas concentrações de Fe, Mn e Zn foi observada quando plantas infectadas e saudáveis foram comparadas.

Alkalithermophilic actinomycetes in a subtropical area of Jujuy, Argentina / Actinomicetos termoalcalófilos del área subtropical de Jujuy, Argentina

The objective of this study was to examine the alkalithermophilic actinomycete communities in the subtropical environment of Jujuy, Argentina, characterized by sugarcane crops. Laceyella putida, Laceyella sacchari, Thermoactinomyces intermedius, Thermoactinomyces vulgaris and Thermoflavimicrobium dichotomicum were isolated on the media with novobiocin, from sugar cane plants and renewal rhizospheres, and grass and wood soils. Soil pH was almost neutral or lightly alkaline, except for grass soil acidified by lactic liquor. A smaller number of actinomycetes was found on the living plants and bagasse (recently obtained or stored according to the Ritter method) with respect to decomposed leaves on the soil. Thermophilic species of Laceyella, Thermoactinomyces, Thermoflavimicrobium, Saccharomonospora, Streptomyces and Thermononospora were isolated on the media without novobiocin, from composted sugar cane residues. Air captured near composted bagasse piles, contained alkalithermophilic actinomycete spores.

El objetivo de este trabajo fue examinar los actinomicetos termoalcalófilos presentes en el área subtropical de Jujuy, Argentina, caracterizada por el cultivo de la caña de azúcar. Se aislaron en medio con novobiocina las especies Laceyella putida, Laceyella sacchari, Thermoactinomyces intermedius, Thermoactinomyces vulgaris y Thermoflavimicrobium dichotomicum a partir de la rizósfera de plantas y de renuevos de caña de azúcar, así como de suelos de pastura y de monte natural. El pH de los suelos era casi neutro a ligeramente alcalino, excepto en un solo caso en que el suelo estaba acidificado por licor láctico. El número de actinomicetos encontrados sobre los tejidos vivos y en el bagazo recién obtenido o almacenado según el método de Ritter fue pequeño en comparación con el observado sobre las hojas en descomposición. L. sacchari predominó respecto de T. vulgaris. Se aislaron especies termoalcalófilas de Laceyella, Thermoactinomyces, Thermoflavimicrobium, Saccharomonospora, Streptomyces y Thermononospora de los residuos compostados de caña de azúcar utilizando medio sin novobiocina. El aire capturado cerca de pilas de bagazo en compostaje contenía esporos de estos organismos.

Influence of roadside pollution on the phylloplane microbial community of Alnus nepalensis (Betulaceae)

The North Eastern region of India is undergoing industrial development at a faster rate than expected. Roads form the main system of transportation and communication owing to the hilly topography of the region. Automobiles discharge a number of gaseous and trace metal contaminants. Human activities like stone grinding, road construction and sand milling also increase the atmospheric dust and heavy metal contaminant level. These contaminants get settled on leaf surfaces at roadsides and enter in contact with phylloplane microorganisms. This study compares microorganisms on leaf surfaces of alder (Alnus nepalensis (Betulaceae)) on roadside and non-roadside environments. Two sites dominated by alder were selected. One at a busy road intersection on the National Highway no. 44 in Shillong with high traffic density (8 000-9 000 heavy vehicles/day), taken as the polluted site and the other one in a forest approximately 500 m away from the roadside considered as the unpolluted site. Analysis of phylloplane microorganisms, lead, zinc, copper, cadmium and sulphur was carried out from leaves. The bacterial population was higher at the unpolluted site. Bacterial population showed a significant negative correlation with lead, zinc, copper, cadmium and sulphur. Similarly, fungal population was higher at the unpolluted site. A total of 29 fungal species were isolated from the phylloplane of A. nepalensis (polluted site 16 species; unpolluted site 28 species). Some fungal forms like Mortierella sp., Fusarium oxysporum and Aureobasidium pollulans were dominant in the polluted site. Numbers of phylloplane fungi and bacteria were significantly reduced in the polluted site. The correlation coefficient indicated a detrimental effect of metals like lead, zinc, copper, cadmium and sulphur on the microbial community of leaf surfaces. The specificity of certain fungi to the unpolluted site may be attributed to their sensitivity to pollution. The predominance of Aureobasidium pollulans, Fusarium oxysporum and Mortierella sp. in the polluted site may be due to their resistance to pollutants in roadside conditions. Rev. Biol. Trop. 56 (3): 1521-1529. Epub 2008 September 30.

La región nororiental de la India está experimentando un rápido desarrollo industrial y debido a la topografía montañosa de la región, los automóviles descargan variedad de contaminantes. Actividades humanas como moler piedra y construir caminos también aumentan el polvo atmosférico y la contaminación con metales pesados. Este estudio compara microorganismos en superficies foliares del árbol Alnus nepalensis (Betulaceae). Se estudió uno sitio en una intersección de Shillong, con alta densidad de tráfico (8 000-9 000 vehículos pesados/día), tomada como el sitio contaminado, y otro en un bosque aproximadamente a 500 m de la carretera (sitio no contaminado). Se contabilizaron bacterias, hongos, plomo, zinc, cobre, cadmio y sulfuro. La población bacteriana era más alta en el sitio no contaminado. La población bacteriana mostró una correlación negativa significativa con el contenido de plomo, cinc, cobre, cadmio y sulfuro foliares en el sitio contaminado, mientras que en el sitio no contaminado la correlación fue insignificante. La población de hongos también fue más alta en el sitio no contaminado. Un total de 29 especies de hongos fueron aislados A. nepalensis: 16 en el sitio contaminado y 28 en el no contaminado. Algunas especies de hongos como Mortierella sp., Fusarium oxysporum y Aureobasidium pollulans fueron dominantes en el sitio contaminado. El número de hongos y bacterias fue significativamente menor en el sitio contaminado. El coeficiente de correlación indicó un efecto perjudicial de metales como el plomo, el zinc, el cobre, el cadmio y el sulfuro en esta comunidad que habitan superficies de hojas. La especificidad de ciertos hongos al sitio no contaminado se puede atribuir a su sensibilidad a la contaminación. El predominio de Aureobasidium pollulans, Fusarium oxysporum y Mortierella sp. en el sitio contaminado puede ser debido a su resistencia a los agentes contaminadores.

Microbial flora associated with submerged mangrove leaf litter in India

We studied the microbial flora in decomposing mangrove leaves in relation to changes in nitrogen and tannin levels, and in penaeid prawn assemblages. Senescent leaves of two mangrove species (Rhizophora apiculata and Avicennia marina) kept in nylon bags, were separately immersed for 80 days in five tanks full of mangrove water. A known amount of decomposing leaves was collected every ten days from each tank for microorganism counts, total nitrogen and tannin measurement, and juvenile penaeid prawn counts. Five genera of total heterotrophic bacteria (THB), three species of azotobacters and 19 species of fungi were identified. The azotobacters showed a significant peak around 40-50 days after the beginning of of decomposition, similar to the trend for total nitrogen and for prawn assemblages. Rev. Biol. Trop. 55 (2): 393-400. Epub 2007 June, 29.

Se estudió la flora microbiana en hojas en descomposición de mangles, considerando nitrógeno, taninos y camarones peneidos jóvenes. Colocamos hojas viejas de dos especies de mangle (Rhizophora apiculata y Avicennia marina) en bolsas de nylon y las sumergimos en agua de manglar durante 80 días usando cinco tanques separados. Cada diez días extrajimos una cantidad conocida de hojas en descomposición de cada tanque. Hallamos cinco géneros de bacterias heterotróficas totales (THB), tres especies de azotobacterias y 19 especies de hongos. Las azotobacterias presentaron un pico significativo de abundancia alrededor de los 40-50 días de descomposición, un patrón similar a los del nitrógeno total y los camarones.

Bacillus thuringiensis in caterpillars and associated materials collected from protected tropical forests in northwestern Costa Rica

Bacillus thuringiensis (Bt) synthesizes crystalline inclusions that are toxic to caterpillars (Lepidoptera) and other orders of invertebrates. Materials associated with 37 caterpillars from 16 species, collected while feeding on 15 different species of host plants in dry, cloud and rain forests located in the Area de Conservación Guanacaste in northwestern Costa Rica, were examined for the presence of Bt. From a total of 101 derived samples, 25 Bt isolates were cultured: 56% from host plant leaves, 8% from caterpillar guts and 36% from caterpillar fecal pellets. Bt was isolated from at least one sample in 38% of the systems constituted by the food plant, gut and fecal pellets corresponding to a single caterpillar. Four different morphologies of crystalline inclusions were observed, with bipyramidal and irregular crystal morphologies being the most prevalent.

Bacillus thuringiensis (Bt) sintetiza inclusiones cristalinas que resultan tóxicas para algunas larvas de lepidópteros y otros órdenes de invertebrados. Su presencia fue examinada en materiales asociados a 37 orugas de mariposas de 16 especies, las cuales fueron colectadas mientras se alimentaban en 15 especies diferentes de plantas hospederas en bosques secos, nubosos y húmedos localizados dentro del Área de Conservación Guanacaste (ACG) en el noroeste de Costa Rica. A partir de un total de 101 muestras se obtuvo 25 aislamientos de Bt: 56% a partir de material foliar de las plantas hospederas, 8% a partir del contenido intestinal de las larvas y 36% a partir de sus excrementos. Esta bacteria fue cultivada a partir de al menos uno de los 3 diferentes tipos de muestra asociados a una oruga particular (planta hospedera, intestino, excremento) en 38% de los casos posibles. En la colección de aislamientos obtenida se observaron cuatro morfologías de inclusiones cristalinas, siendo aquellas bipiramidales e irregulares las más prevalentes.