In vitro effect of substrate, temperature and photoperiod on Phakopsora pachyrhizi urediniospore germination and germ tube growth

In vitro experiments were conducted to assess the effects of substrate, temperature and time of exposure to temperature and photoperiod on P. pachyrhizi uredospore germination and germ tube growth. The following substrates were tested: water-agar and soybean leaf extract-agar at different leaf concentrations (0.5, 1.0, 2.0 and 4.0 g of leaves and 15g agar/L water), temperatures (10, 15, 20, 25, 30, and 35oC) and times of exposure (1, 2, 3, 4, 5, 6, 7, and 8 hours) to temperature and 12 different photoperiods. The highest germination and germ tube length was found for the soybean leaf extract agar. Maximum P. pachyrhizi uredospore germination was obtained at 21.8 and 22.3°C, and maximum germ tube growth at 21.4 and 22.1°C. The maximum uredospore germination was found at 6.4 hours exposure, while the maximum germ tube length was obtained at 7.7 h exposure. Regarding photoperiod, the maximum spore germination and the maximum uredospore germ tube length were found in the dark. Neither spore germination nor uredospore germ tube growth was completely inhibited by the exposure to continuous light.

Harvesting and wood transport planning with SNAP III program (Scheduling and Network Analysis Program) in a pine plantation in Southeast Brazil

The objective of this study was to verify the potential of SNAP III (Scheduling and Network Analysis Program) as a support tool for harvesting and wood transport planning in Brazil harvesting subsystem definition and establishment of a compatible route were assessed. Initially, machine operational and production costs were determined in seven subsystems for the study area, and quality indexes, construction and maintenance costs of forest roads were obtained and used as SNAP III program input data. The results showed, that three categories of forest road occurrence were observed in the study area: main, secondary and tertiary which, based on quality index, allowed a medium vehicle speed of about 41, 30 and 24 km/hours and a construction cost of about US$ 5,084.30, US$ 2,275.28 and US$ 1,650.00/km, respectively. The SNAP III program used as a support tool for the planning, was found to have a high potential tool in the harvesting and wood transport planning. The program was capable of defining efficiently, the harvesting subsystem on technical and economical basis, the best wood transport route and the forest road to be used in each period of the horizon planning.

In vitro EVALUATION OF EUCALYPTUS ECTOMYCORRHIZAE ON SUBSTRATE WITH PHOSPHORUS DOSES FOR FUNGAL PRE-SELECTION

The benefit promoted by ectomycorrhizal depends on the interaction between symbionts and phosphorus (P) contents. Phosphorus effect on ectomycorrhizal formation and the effectiveness of these in promoting plant growth for fungal pre-selection were assessed under in vitro conditions. For P effect evaluation, Eucalyptus urophylla seedlings inoculated with four Pisolithus sp. isolates and others non-inoculated were grown on substrate containing 0.87, 1.16 and 1.72 mg P per plant. For evaluation of effectiveness and fungal pre-selection, other 30 isolates of Pisolithus sp., Pisolithus microcarpus ITA06 isolate, Amanita muscaria AM16 isolate, Scleroderma areolatum SC129 isolate were studied. D26 isolate promoted the highest plant heights for the three P doses, D51 at the lower dose and D72 at the intermediate dose. P doses did not influenced shoot fresh weight and fungal colonization. In the pre-selection of fungi, 14 isolates of Pisolithus sp., P. microcarpus ITA06 isolate and S. areolatum SC129isolate increased plant height and fresh weight. D82 isolate of Pisolithus sp. had effect singly on plant height while D17 and D58 on fresh weight. Of these, only D15, D17, D58 and ITA06 had typical ectomycorrhizae. The cultivation in vitro has shown adequate for pre-selection of ectomycorrhizal fungi. Colonization and benefits depend on species and isolate. D15, D17 and D58 of Pisolithus sp. and P. microcarpus isolate ITA06 are the most promising for nursery studies.

TEMPERATURE AND SUBSTRATE INFLUENCES ON SEED GERMINATION AND SEEDLING FORMATION IN Callisthene fasciculata Mart. (VOCHYSIACEAE) IN THE LABORATORY1

ABSTRACTCallisthene fasciculata Mart. is a tree belonging to the Vochysiaceae family. Its wood is moderately heavy and resistant and used to make poles, beams, and other structures. The aim of this work was to evaluate seed germination and the initial growth of seedlings of C. fasciculata at different temperatures and in different substrates. Seeds were collected from fruits in the Pantanal de Miranda, Mato Grosso do Sul state, Brazil. In one experiment, the seeds were subjected to constant temperatures of 20, 25, 30 and 35 °C and to alternating temperatures of 20-30 and 25-35 °C (on paper substrate). In another experiment, the seeds were subjected to temperatures of 20 and 25 °C on three substrates (sand, vermiculite and between paper) in a germinator. The experiment had a randomized design, with four replicates of 25 seeds per treatment. The F-values obtained for germination indicated no significant effect of substrate or temperature on the final germination percentage. The analyses revealed no effect of a treatment interaction (temperature x substrate) on either germination or average germination time; however, a treatment interaction effect was observed on the germination speed index. The treatment combinations yielding the best performance were between paper substrate at 20 °C and sand substrate at 25 °C. There was a significant effect of the interaction between temperature and substrate on seedling growth, with increased root growth observed in the between paper substrate at 25 °C and increased aerial component growth in both sand at 20 °C and vermiculite at 25 °C. The between paper treatment at 25 °C yielded the greatest final seedling size. Between paper is the most recommended substrate for the production of seedlings due to its ease of handling and lower probability of contamination.

SUBSTRATE, LIME, PHOSPHORUS AND TOPDRESS FERTILIZATION IN MACAW PALM SEEDLING PRODUCTION

ABSTRACT The macaw palm [Acrocomia aculeata (Jacq.) Lood. ex Mart] has been domesticated to subsidize biodiesel production programs in Brazil. However, little is known about the seedling production of this species. This study aimed to evaluate substrate mixtures, limestone and phosphorus rates for substrate amendment and topdressing frequency in macaw palm seedlings. Three trials were conducted in a greenhouse up to six months of nursery cultivation. Trial 1: determination of percent mineral and organic fractions of seven substrate mixtures. Trial 2: evaluation of four limerates for soil amendment versus four phosphorus rates. Trial 3: evaluation of N, K and Mg topdressing frequency. Significant differences were found in the three trials for most of the variables (plant height, leaf number, shoot dry mass, root dry mass, vigor and bulb diameter). The main results obtained were as follow: Trial1 - the best seedling growth was observed in substrates with at least 25% organic matter. Trial2 -lime rates ranging from 0.50 to 1.25 kg associated with 3 to 4 kg of single superphosphate per m3 of substrate provided the best seedling growth. Trial 3 - topdressing fertilization provided better development of seedlings regardless of frequency.

Temperature effect in potassium and nitrate ions in soil transport

When doing researches on solute dynamics in porous medium, the knowledge of medium characteristics and percolating liquids, as well as of external factors is very important. An important external factor is temperature and, in this sense, our purpose was determining potassium and nitrate transport parameters for different values of temperature, in miscible displacement experiments. Evaluated parameters were retardation factor (R), diffusion/dispersion coefficient (D) and dispersivity, at ambient temperature (25 up to 28 ºC), 40 ºC and 50 ºC. Salts used were potassium nitrate and potassium chlorate, prepared in a solution made up of 5 ppm nitrate and 2.000 ppm potassium, with Red-Yellow Latosol porous medium. Temperature exhibited a positive influence upon porous medium solution and upon dispersion coefficient.

Substrate with Organosuper® for cucumber seedlings formation in protected environments and polystyrene trays

Culture environments, trays and doses of organic compost were evaluated in the formation of cucumber seedlings (Cucumis sativus L.). Five environmental conditions were tested: (A1) a greenhouse with height of 2.5 m, covered with polyethylene film, (A2) nursery with height of 2.5 m, monofilament fabric, 50% shading, (A3) nursery with height of 2.5 m, heat-reflective screen, 50% shading, (A4) nursery with a height of 1.8 m, covered with coconut tree straw and (A5) greenhouse with height of 4.0 m, covered with polyethylene film, with zenith opening and thermo-reflective cloth under the plastic. Trays of 72 (R1) and 128 (R2) cells were filled with 93% soil and 7% organic compound (S1), 86% soil and 14% organic compound (S2) and 79% soil and 21% organic compound (S3). It was used a randomized design in split-split-plot scheme, with five replicates (environments x trays x substrates). The greenhouses provide the best environments for the formation of cucumber seedlings. A tray of 72 cells is the best container, promoting more vigorous seedlings in substrate with soil and 7 or 14% organic compound.

Transport of broilers: load microclimate during Brazilian summer

The objective of this study was to characterize the microclimatic profile of broiler transport trucks under commercial conditions, in the summer, by continuous monitoring of environmental variables (temperature and air relative humidity). Three loads were monitored from farm to slaughterhouse, considering factors such as distance and day periods (morning, afternoon, and night). To obtain the profile of the environmental variables during journey, data loggers, that determined the microclimate to which birds were submitted, were installed in the trucks; data loggers also allowed visualization of the Enthalpy Comfort Index (ECI) so that load regions could be classified according to heat comfort limits for 6-week old poultries. Temperature, relative humidity, and ECI in the truck were analyzed, using geostatistics, by the ordinary kriging method. The afternoon was the most critical period, and truck central and rear regions were most problematic for chickens, thus most susceptible to losses.

Effect of companion ion on nitrate displacement through transport parameters analysis

Nitrate is the main form of nitrogen associated with water contamination; the high mobility of this species in soil justifies the concern regarding nitrogen management in agricultural soils. Therefore, the objective of this research was to assess the effect of companion cation on nitrate displacement, by analyzing nitrate transport parameters through Breakthrough Curves (BTCs) and their settings made by numerical model (STANMOD). The experiment was carried out in the Soil and Water Quality Laboratory of the Department of Biosystems Engineering, "Luiz de Queiroz" College of Agriculture in Piracicaba (SP), Brazil. It was performed using saturated soil columns in steady-state flow condition, in which two different sources of inorganic nitrate Ca(NO3)2 and NH4NO3 were applied at a concentration of 50 mg L-1 NO3-. Each column was filled with either a Red-Yellow Oxisol (S1) or an Alfisol (S2). Results are indicative that the companion ion had no effect on nitrate displacement. However, nitrate transport was influenced by soil texture, particle aggregation, solution speed in soil and organic matter presence. Nitrate mobility was higher in the Alfisol (S2).

Spatial distribution and preferential substrate of Neoregelia johannis (Carriére) L.B. Smith (Bromeliaceae) in a disturbed area of Atlantic Rainforest at Ilha Grande, RJ, Brazil

In bromeliad populations, amount of light and available substrates influence individuals spatial organization. In Atlantic Rainforest of Ilha Grande, the heliophylous bromeliad Neoregelia johannis is a large and abundant species. In this forest, it would be expected that N. johannis would occupy stable substrates, as large trunks, large branches, rock boulders or ground, with high sunlight, enough for the bromeliad survivor. In the present work, we analyzed the distribution and most used substrates of N. johannis in secondary forest. We analyzed the frequency of reproductive modes (sexual and vegetative) used by the bromeliad shoots, registering if the shoots were originated from seeds or by vegetative reproduction. The results indicated an aggregated distribution pattern (Ip = 0.052). The preferred substrate was boulders (91%), whereas tree trunks (6%) and the ground (3%) were rarely used. Small and fragile substrates are unstable to support large adults of this species, which may explain the predominant pattern of establishment over boulders within the secondary forest, as the presence of this substrate also results in more opened canopy cover. Approximately 50% of young individuals entered the population by vegetative reproduction. We conclude that the preferential habit and the aggregated distribution of N. johannis are due to the conjunction of preferred substrate with higher amount of light resulting from breaks in tree canopy over areas with rock blocks, and high frequency of recruitment by vegetative reproduction.

The insulin receptor substrate 1 associates with phosphotyrosine phosphatase SHPTP2 in liver and muscle of rats

Insulin stimulates the tyrosine kinase activity of its receptor resulting in the phosphorylation of its cytosolic substrate, insulin receptor substrate-1 (IRS-1) which, in turn, associates with proteins containing SH2 domains. It has been shown that IRS-1 associates with the tyrosine phosphatase SHPTP2 in cell cultures. While the effect of the IRS-1/SHPTP2 association on insulin signal transduction is not completely known, this association may dephosphorylate IRS-1 and may play a critical role in the mitogenic actions of insulin. However, there is no physiological demonstration of this pathway of insulin action in animal tissues. In the present study we investigated the ability of insulin to induce association between IRS-1 and SHPTP2 in liver and muscle of intact rats, by co-immunoprecipitation with anti-IRS-1 antibody and anti-SHPTP2 antibody. In both tissues there was an increase in IRS-1 association with SHPTP2 after insulin stimulation. This association occurred when IRS-1 had the highest level of tyrosine phosphorylation and the decrease in this association was more rapid than the decrease in IRS-1 phosphorylation levels. The data provide evidence against the participation of SHPTP2 in IRS-1 dephosphorylation in rat tissues, and suggest that the insulin signal transduction pathway in rat tissues is related mainly to the mitogenic effects of the hormone.

Erythrocyte glucose-6-phosphate dehydrogenase activity assay and affinity for its substrate under "physiological" conditions

Glucose-6-phosphate dehydrogenase (G6PD) activity and the affinity for its substrate glucose-6-phosphate were investigated under conditions similar to the physiological environment in terms of ionic strength (I: 0.188), cation concentration, pH 7.34, and temperature (37oC). A 12.4, 10.4 and 21.4% decrease was observed in G6PD B, G6PD A+ and G6PD A- activities, respectively. A Km increase of 95.1, 94.4 and 95.4% was observed in G6PD B, G6PD A+ and G6PD A-, respectively, leading to a marked decrease in affinity. In conclusion, the observation of the reduced activity and affinity for its natural substrate reflects the actual pentose pathway rate. It also suggests a much lower NADPH generation, which is crucial mostly in G6PD-deficient individuals, whose NADPH availability is poor.

P-glycoprotein-mediated resistance to chemotherapy in cancer cells: using recombinant cytosolic domains to establish structure-function relationships

Resistance to chemotherapy in cancer cells is mainly mediated by overexpression of P-glycoprotein (Pgp), a plasma membrane ATP-binding cassette (ABC) transporter which extrudes cytotoxic drugs at the expense of ATP hydrolysis. Pgp consists of two homologous halves each containing a transmembrane domain and a cytosolic nucleotide-binding domain (NBD) which contains two consensus Walker motifs, A and B, involved in ATP binding and hydrolysis. The protein also contains an S signature characteristic of ABC transporters. The molecular mechanism of Pgp-mediated drug transport is not known. Since the transporter has an extraordinarily broad substrate specificity, its cellular function has been described as a "hydrophobic vacuum cleaner". The limited knowledge about the mechanism of Pgp, partly due to the lack of a high-resolution structure, is well reflected in the failure to efficiently inhibit its activity in cancer cells and thus to reverse multidrug resistance (MDR). In contrast to the difficulties encountered when studying the full-length Pgp, the recombinant NBDs can be obtained in large amounts as soluble proteins. The biochemical and biophysical characterization of recombinant NBDs is shown here to provide a suitable alternative route to establish structure-function relationships. NBDs were shown to bind ATP and analogues as well as potent modulators of MDR, such as hydrophobic steroids, at a region close to the ATP site. Interestingly, flavonoids also bind to NBDs with high affinity. Their binding site partly overlaps both the ATP-binding site and the steroid-interacting region. Therefore flavonoids constitute a new promising class of bifunctional modulators of Pgp.

Activity and functional interaction of alternative oxidase and uncoupling protein in mitochondria from tomato fruit

Cyanide-resistant alternative oxidase (AOX) is not limited to plant mitochondria and is widespread among several types of protists. The uncoupling protein (UCP) is much more widespread than previously believed, not only in tissues of higher animals but also in plants and in an amoeboid protozoan. The redox energy-dissipating pathway (AOX) and the proton electrochemical gradient energy-dissipating pathway (UCP) lead to the same final effect, i.e., a decrease in ATP synthesis and an increase in heat production. Studies with green tomato fruit mitochondria show that both proteins are present simultaneously in the membrane. This raises the question of a specific physiological role for each energy-dissipating system and of a possible functional connection between them (shared regulation). Linoleic acid, an abundant free fatty acid in plants which activates UCP, strongly inhibits cyanide-resistant respiration mediated by AOX. Moreover, studies of the evolution of AOX and UCP protein expression and of their activities during post-harvest ripening of tomato fruit show that AOX and plant UCP work sequentially: AOX activity decreases in early post-growing stages and UCP activity is decreased in late ripening stages. Electron partitioning between the alternative oxidase and the cytochrome pathway as well as H+ gradient partitioning between ATP synthase and UCP can be evaluated by the ADP/O method. This method facilitates description of the kinetics of energy-dissipating pathways and of ATP synthase when state 3 respiration is decreased by limitation of oxidizable substrate.

Nonconventional amide bond formation catalysis: programming enzyme specificity with substrate mimetics

This article reports on the design and characteristics of substrate mimetics in protease-catalyzed reactions. Firstly, the basis of protease-catalyzed peptide synthesis and the general advantages of substrate mimetics over common acyl donor components are described. The binding behavior of these artificial substrates and the mechanism of catalysis are further discussed on the basis of hydrolysis, acyl transfer, protein-ligand docking, and molecular dynamics studies on the trypsin model. The general validity of the substrate mimetic concept is illustrated by the expansion of this strategy to trypsin-like, glutamic acid-specific, and hydrophobic amino acid-specific proteases. Finally, opportunities for the combination of the substrate mimetic strategy with the chemical solid-phase peptide synthesis and the use of substrate mimetics for non-peptide organic amide synthesis are presented.