Cloning, expression, purification and characterization of recombinant glutathione-S-transferase from Xylella fastidiosa

Xylella fastidiosa is an important pathogen bacterium transmitted by xylem-feedings leafhoppers that colonizes the xylem of plants and causes diseases on several important crops including citrus variegated chlorosis (CVC) in orange and lime trees. Glutathione-S-transferases (GST) form a group of multifunctional isoenzymes that catalyzes both glutathione (GSH)-dependent conjugation and reduction reactions involved in the cellular detoxification of xenobiotic and endobiotic compounds. GSTs are the major detoxification enzymes found in the intracellular space and mainly in the cytosol from prokaryotes to mammals, and may be involved in the regulation of stress-activated signals by suppressing apoptosis signal-regulating kinase 1. In this study, we describe the cloning of the glutathione-S-transferase from X. fastidiosa into pET-28a(+) vector, its expression in Escherichia coli, purification and initial structural characterization. The purification of recombinant xfGST (rxfGST) to near homogeneity was achieved using affinity chromatography and size-exclusion chromatography (SEC). SEC demonstrated that rxfGST is a homodimer in solution. The secondary and tertiary structures of recombinant protein were analyzed by circular dichroism and fluorescence spectroscopy, respectively. The enzyme was assayed for activity and the results taken together indicated that rxfGST is a stable molecule, correctly folded, and highly active. Several members of the GST family have been extensively studied. However, xfGST is part of a less-studied subfamily which yet has not been structurally and biochemically characterized. In addition, these studies should provide a useful basis for future studies and biotechnological approaches of rxfGST. (C) 2008 Elsevier B.V. All rights reserved.

Flooding tolerance of Tabebuia cassinoides: Metabolic, morphological and growth responses

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The protist Trichomonas vaginalis harbors multiple lineages of transcriptionally active Mutator-like elements

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of calcium and magnesium silicate on the growth of the castor oil plant subjected to salinity levels

Salt stress decreases the osmotic potential of soil solution causing water stress, causing toxic effects in the plants resulting in injuries on the metabolism and nutritional disorders, thus compromising the plant growth, resulting in lower production. The calcium silicate and magnesium can perform the same function as limestone, besides providing silicon to plants, may also contribute to the resistance of plants to salt stress. Thus, the objective of this study was to evaluate the effect of calcium and magnesium silicate on the growth of the castor oil plant BRS Energia cultivated under saline conditions. This study evaluated plant height, stem diameter, number of leaves, leaf area, dry weight of shoot and root, and soil chemical characteristics. There was no interaction between factors of salinity level and of silicate level regarding the evaluated variables. There was a direct relationship between salinity levels and plant growth in height and stem diameter. The K concentration in soil were affected by salinity levels.

Hepatogenous photosensitization of ruminants by Brachiaria decumbens and Panicum dichotomiflorum in the absence of sporidesmin: Lithogenic saponins may be responsible

As part of a study of plants involved in crystal-associated hepatogenous photosensitization diseases, samples of Brachiaria decumbens and Panicum dichotomiflorum on which cattle and goats had recently been photosensitized were analyzed. The level of saponins associated with these photosensitization outbreaks were determined by GC-MS. Only low levels of Pithomyces chartarum spores were present on the B decumbens, and all isolates obtained failed to produce sporidesmin.

Seasonal variation in coumarin content of Mikania glomerata

Mikania glomerata Spengal extract contains approximately 0.5 percent of coumarin, a substance that displays anti-inflammatory and expectoratory activities. Extracts from different young leaves of plants collected during the early evening of December and July contained the highest levels coumarin. Plants dried in oven with circulating air contained 7.3 mg of coumarin/g of dry weight, demonstrating that this was the most appropriate drying condition for Mikania glomerata.

Antifungal activity of propolis on different species of Candida

Propolis is a resinous material collected by bees from the buds or other parts of plants. It is known for its biological properties, having antibacterial, antifungal and healing properties. The antifungal activity of propolis was studied in sensitivity tests on 80 strains of Candida yeasts: 20 strains of Candida albicans, 20 strains of Candida tropicalis, 20 strains of Candida krusei and 15 strains of Candida guilliermondii. The yeasts showed a clear antifungal activity with the following order of sensitivity: C. albicans > C. tropicalis > C. krusei > C. guilliermondii. Patients with full dentures who used a hydroalcoholic propolis extract showed a decrease in the number of Candida.

Taxonomy and distribution of Paralemanea (Lemaneaceae, Rhodophyta) in Central Mexico

Three collections of Paralemanea from Central Mexico included two species. Paralemanea mexicana is large (length ≥ 4.0 cm; diameter > 400 μm) and generally branched (≥ 40 % of plants branched), with whorled branches, of first to second order. Paralemanea annulata is small (length < 5.0 cm ; diameter < 500 μm), generally unbranched (≤ 5 % of plants branched), with branches of first order. Spermatangial sori contained obovoid spermatangia, formed from cells of the outer cortical layers, extending above the thallus surface. Carpogonial branches are described for the first time in P. mexicana. They develop on lateral filaments at nodes or internodes and have ovoid to globular cells, abundantly branched at the basal portion, penetrating the cortex towards the thallus surface. Carposporophytes are sessile on the inner portion of the cortex and produce carpospores in chains of up to twelve. The 'Chantransia' stage was observed in P. mexicana. Paralemanea annulata is described for the first time from Mexico and P. mexicana is endemic from this country. Both species were collected in cold (temperature 12-16°C), acidic (pH 5.5-6.0), shallow (depth 1-60 cm) and moderate to fast flowing waters (> 35 cm s-1), in shaded or partly shaded river segments, on rocky substrata (mostly bedrock).

Termite pests in eucalyptus forests of brazil

The termite problem in eucalyptus forest plantations in Brazil has been registered since 1908. The main termite pests can be separated in four groups: a) seedling/sapling termites; b) heartwood termites; c) bark termites and d) wood termites. The termites in the first group attack root and stalk bases of young eucalyptus plants. The most common species are Syntermes spp. and Cornitermes spp. and they are a serious obstacle to early eucalyptus developing. The heartwood termites attack formed trees destroying eucalyptus heartwood. Coptotermes testaceus is the most cited species in reports, but more species probably occur. Plant mortality caused by seedling/sapling termites vary of 10-70 %. There are not effective control methods to heartwood termites. The main seedling/sapling termite control strategy is the chemical barrier around root systems of plants. Nowadays, studies are being carried out to determine monitoring systems to termite infestations. Early results indicate that proportionally, few areas really need insecticide application, due to spatial distribution of termites to be aggregated. Therefore, it is necessary to develope techniques rationalizing insecticide utilization in eucalyptus plantations, to keep production systems feasible and to be suitable for environmental exigencies.

Collection of Fáfia [Pfaffia glomerata (Spreng.) Pedersen] in Northwestern State of Paraná - Brazil

Interviews were conducted with fáfia (Pfaffia glomerata (Spreng.) Pedersen) collectors from the municipal districts of Querência do Norte and Porto Rico, where enterprises and buyers of this plant are concentrated. The results allowed us to conclude that both the adults and children make collection. Each adult collects from 50 to 150 kg of roots/day, depending on the collection area, for about 8 months/year. Collections mostly occur all over the year, but the activity becomes more intense from May to August. All families are not exclusively dedicated to fáfia collection and also develop other rural activities. They have been collecting fáfia for 2 to 13 years, indicating that an intensive exploitation has been present in the region for over a decade. During collection, no plant part is used for replanting the species. The roots are commercialised by regional buyers. The price of the roots varies from US $ 0.07 to US $ 0.13/kg and average gain is about US $ 2,055/family/year, representing a considerable profit for the collectors.

The effect of plant diversity on fungus garden development and foraging behavior of leaf-cutting ants (Hymenoptera: Formicidae)

The leaf-cutting ants forage a wide variety of plant species, used for symbiotic fungus cultivation. To better understand this tripartite complex interaction, 24 colonies of Acromyrmex subterraneus brunneus were conditioned for 4 months to 6 different plants (Citrus spp., Ligustrum spp., Acalypha spp., Eucalyptus spp., Alchornea triplinervia, Melia spp.), to verify the influence of conditioning on foraging behavior of workers. The effect of plants on symbiotic fungus development was studied separately, through macerated plants in Agar and culture medium A as the control. During foraging, workers presented polyphagic foraging behavior, refusing the plants to which they were conditioned. The selection of plants is not correlated with the plant substrate that promotes good development of symbiotic fungus. Such results demonstrate the importance of plant diversity for fungus garden maintenance.

Growth and yield analysis in different methods of beet (Cv. 'tall top early wonder') plantation

This objective of this study was to evaluate the effects of different methods of red beet seedling production and direct sowing on the development of the plant in field conditions. 'Tall Top Early Wonder' was used as the cultivar. The experimental design was a randomized blocks with 4 replications of each treatment: T1 - seedling produced in trays of 288 cells; T2 - 200 cells; T3 - 128 cells; T4 - 128 cells; T5 - direct sowing and T6 - bare-root seedling. The seedlings produced in trays and bare root seedlings were transplanted in the field 28 DAS with spacing of 0.20 x 0.10m. The mean height of plants, leaf area, leaf dry matter, petiole dry matter, shoot dry matter, root dry matter, relationship between shoot dry matter and root dry matter, absolute growth rate, relative growth rate, net assimilation rate, leaf area ratio and specific leaf area were determined. Storage root fresh matter and mean storage root diameter were determined starting from 77 DAS. Initial growth of the plants were superior for the direct sowing, resulting in smaller RGR and NAR than the other treatments. Independent of the production method, an increase of the cycle of the crop was verified. T6 had larger delay in the initial development. There was no difference for productivity. Method T1, had less expenses with respect to substrate and space in the vegetation home, without reduction in production.

Differential fruit consumption of two Melastomataceae by birds in Serra da Mantiqueira, southeastern Brazil

In this study we report on the consumption of two syntopic Melastomataceae species by birds in a lower montane forest in Monte Verde, southeastern Brazil. The species of frugivores were identified and characterized by their methods of capture and consumption of fruits. We also provide information on abundance, phenology of plants and fruit characteristics of the two Melastomataceae species. The 13 observed species of birds formed two statistically distinct frugivorous groups with taxonomic and behavioral differences. Five of seven bird species that fed on L. aff. sublanata fruits belong to the subfamily Thraupinae and most fruits were mashed before swallowed. Four of the eight bird species that visited M. cinerascens belong to the subfamily Turdinae and all fruits were swallowed whole. Only two bird species were common visitors of both Melastomataceae species. Our findings show that fruits of the two Melastomataceae species with similar morphological characteristics were exploited differently by frugivorours birds.

Interference of epicuticular wax from leaves of grasses in selection and preparation of substrate for cultivation of symbiont fungus by Atta capiguara (Hym. Formicidae)

The selection of plants is one of the stages of foraging behavior, executed by leaf-cutting ants for the cultivation of symbiont fungus. Anatomical, biochemical and physiological characteristics of vegetal species are some of the factors that influence selection of substrate; however, most studies described in the literature refer to leaf-cutting ants. Due to the scarcity of studies on the role of epicuticular waxes of grasses in relation to foraging behavior of monocotyledonous leaf-cutters, we completed three experiments with the ant species Atta capiguara in which we analyzed: the attractiveness or selectivity of leaves of Saccharum officinarum (sugarcane) and/or Hypharrenia rufa (capim jaraguá), with and without epicuticular wax in relation to the worker of this ant species, their processing and elimination in the form of pellets of epicuticular wax from the referred fragments removed by the workers for cultivation of symbiont fungus. We observed that vegetal fragments without wax were those most transported by workers to the inside of the nests; that they processed the fragments equally with as much as without wax, and eliminated, in the form of pellets, wax removed from the same into garbage chambers of the colonies, leading us to conclude that for monocotyledonous leaf-cutters, epicuticular wax from vegetal leaves has great importance in vegetal integrity by being used as substrate, representing a physical barrier to the growth of symbiont fungus.

Eucalyptus ESTs associated with resistance to herbicide inhibitors of aromatic and branched-chain amino acid synthesis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)