Extraction and concentration of biogenic calcium oxalate from plant leaves

The objective of this study was to extract and concentrate calcium oxalate (CaOx) crystals from plant leaves that form the above mentioned crystals. The chemical and physical studies of CaOx from plant to be performed depend on an adequate amount of the crystals. The plant used in this study was croton (Codiaeum variegatum). The leaves were ground in a heavy duty blender and sieved through a 0.20 mm sieve. The suspension obtained was suspended in distilled water. The crystals were concentrated at the bottom of a test tube. The supernatant must be washed until it is free of plant pigments and other organic substances. Biogenic CaOx crystals have well-defined and sharp peaks, indicating very high crystallinity. Moreover, the CaOx crystals were not damaged during the extraction procedure, as can be seen on the scanning electron microscope images. The porposed method can be considered efficient to extract and concentrate biogenic calcium oxalate.

Boron extraction and vertical mobility in Paraná State oxisol, Brazil

The deficiency or excess of micronutrients has been determined by analyses of soil and plant tissue. In Brazil, the lack of studies that would define and standardize extraction and determination methods, as well as lack of correlation and calibration studies, makes it difficult to establish limits of concentration classes for analysis interpretation and fertilizer recommendations for crops. A specific extractor for soil analysis is sometimes chosen due to the ease of use in the laboratory and not in view of its efficiency in determining a bioavailable nutrient. The objectives of this study were to: (a) evaluate B concentrations in the soil as related to the fertilizer rate, soil depth and extractor; (b) verify the nutrient movement in the soil profile; (c) evaluate efficiency of Hot Water, Mehlich-1 and Mehlich-3 as available B extractors, using sunflower as test plant. The experimental design consisted of complete randomized blocks with four replications and treatments of five B rates (0, 2, 4, 6, and 8 kg ha-1) applied to the soil surface and evaluated at six depths (0-0.05, 0.05-0.10, 0.10-0.15, 0.15-0.20, 0.20-0.30, and 0.30-0.40 m). Boron concentrations in the soil extracted by Hot Water, Mehlich-1 and Mehlich-3 extractors increased linearly in relation to B rates at all depths evaluated, indicating B mobility in the profile. The extractors had different B extraction capacities, but were all efficient to evaluate bioavailability of the nutrient to sunflower. Mehlich-1 and Mehlich-3 can therefore be used to analyze B as well as Hot Water.

Simultaneous multielement extraction with the Mehlich-1 solution for Southern Brazilian soils determined by ICP-OES and the effects on the nutrients recommendations to crops

The amounts of macro (P, K, Ca and Mg) and micronutrients (Cu and Zn) extracted with the Mehlich-1 (M1) solution, by the 1.0 mol L-1 KCl (KCl) and with the 0.1 mol L-1 HCl (HCl) for representative soil types of the Rio Grande do Sul state (Brazil) were compared with those extracted with the Mehlich-1 solution determined with the inductively coupled plasma optical emission spectroscopy (ICP). The amounts of nutrients extracted by the different methods showed high correlation coefficients. On average, the Mehlich-1 solution extracted similar amounts of P, determined with colorimetric and ICP methods, and, K determined with emission and ICP. The amounts of Ca and Mg extracted with the Mehlich-1 solution, determined by ICP, were similar to those extracted with the KCl solution determined by the atomic absorption spectrophotometry. The amounts of Cu and Zn extracted with the Mehlich-1 solution, determined by the ICP, were higher than those extracted with the 0.1 mol L-1 HCl determined by the atomic absorption spectrophotometry. The results indicate that the Mehlich-1 solution and ICP can be used for simultaneous multielement extraction and determination for Southern Brazilian soils. However, a conversion factor for values interpretation is needed. The use of the conversion factor to determine the K availability index in soils is adequate and does not affect the K recommendations for crops in southern Brazilian soils.

Nutrient extraction and exportation by castor bean hybrid lyra

Information about nutrient extraction and exportation by crops, as well as the periods of highest nutrient demand is important for an adequate fertilization management. However, there are no studies on the nutrient uptake of short-stature hybrid castor bean. Therefore, the purpose of this study was to evaluate nutrient extraction and exportation by short-stature castor bean hybrid Lyra, in the spring-summer and fall-winter growing seasons. The experiments were conducted in the 2005/2006 spring-summer and 2006 fall-winter growing seasons on an Oxisol, in Botucatu, SP, in a randomized block design, with four replications. The plots consisted of plant samplings, which occurred 17, 31, 45, 59, 73, 97 and 120 days after emergence (DAE) in the spring-summer and 17, 31, 45, 59, 80, 100 and 120 DAE in fall-winter growing season. The growth of hybrid Lyra was slow and nutrient uptake lowest between emergence and the beginning of flowering. The period of highest dry matter (DM) accumulation rates and highest nutrient demand were observed 40 to 80 DAE, in both growing seasons. The order of nutrient extraction by the plants in the spring-summer growing season was: N>K>Ca>Mg>S>P>Fe>Mn>Zn>B>Cu>Mo. In fall-winter, S was more absorbed than Mg. Seed yield was higher in the spring-summer (2.995 kg ha-1), but nutrient extraction and exportation per ton of seed were similar in both growing seasons. Around 58 % of N and 84 % of P, and approximately half of the S and B absorbed throughout the cycle were exported with the seeds. However, most of the other nutrients accumulated in the plants returned to the soil in plant residues.

Nutrient extraction and exportation by common bean cultivars under different fertilization levels: I - macronutrients

The use of cultivars with a higher yield potential and the adoption of new technology have achieved high grain yields in common bean, which probably changed the demand for nutrients in this crop. However, there is almost no information about the periods of the cycle in which nutrients are most demanded at which quantities by the main cultivars. The objective of this study was to evaluate the macronutrient extraction and exportation by the common bean cultivars Pérola and IAC Alvorada, under different levels of NPK fertilization, on a dystroferric Red Nitosol, in Botucatu, São Paulo State, Brazil. The experiment was arranged in a randomized complete block (split plot) design with four replications. The plots consisted of six treatments based on a 2 x 3 factorial model, represented by two cultivars and three NPK levels (PD0 - 'Pérola' without fertilization, PD1 - 'Pérola' with 50 % of recommended fertilization, PD2 - 'Pérola' with 100 % of recommended fertilization, AD0 - 'IAC Alvorada' without fertilization, AD1 - 'IAC Alvorada' with 50 % of recommended fertilization, and AD2 - 'IAC Alvorada' with 100 % of recommended fertilization) and subplots sampled seven times during the cycle. At higher levels of NPK fertilization, the grain yield and macronutrient extraction and exportation of both cultivars were higher, but without statistical differences. Macronutrient absorption was higher in the treatments with 100 % of recommended NPK fertilization (average amounts per hectare: 140 kg N, 16.5 kg P, 120 kg K, 69 kg Ca, 17.9 kg Mg, and 16.3 kg S). Regardless of the treatment, the demand for N, P, K, Ca, and Mg was highest from 45 to 55 days after emergence (DAE), i.e., in the R7 stage (pod formation), while the highest S absorption rates were concentrated between 55 and 65 DAE. More than 70 % of P, between 58 and 69 % of N, 40 and 52 % of S, 40 and 48 % of K, and 35 and 45 % of Mg absorbed during the cycle was exported with grains, whereas less than 15 % of Ca was exported.

Nutrient extraction and exportation by common bean cultivars under different fertilization levels: II - micronutrients

Where the level of agricultural technology is higher, common bean cultivars with a higher yield potential possibly require greater amounts of micronutrients. In Brazil however, there is a lack of information about the micronutrient extraction and exportation by the main grown cultivars. The objective of this study was to evaluate micronutrient (B, Cu, Fe, Mn, and Zn) extraction and exportation by common bean cultivars Pérola and IAC Alvorada, under different levels of NPK fertilization, on a dystroferric Red Nitosol, in Botucatu, São Paulo State, Brazil. The experiment was arranged in a randomized complete block (split plot) design with four replications. The plots consisted of six treatments based on a 2 x 3 factorial model, represented by two cultivars and three NPK levels (PD0 - 'Pérola' without fertilization, PD1 - 'Pérola' with 50 % of recommended fertilization, PD2 - 'Pérola' with 100 % of recommended fertilization, AD0 - 'IAC Alvorada' without fertilization, AD1 - 'IAC Alvorada' with 50 % of recommended fertilization, and AD2 - 'IAC Alvorada' with 100 % of recommended fertilization) and subplots sampled seven times during the cycle. Higher levels of NPK fertilization increased micronutrient extraction by both cultivars, and treatments with 100 % of recommended NPK fertilization extracted on average 167 g B, 58 g Cu, 1,405 g Fe, 1,213 g Mn and 211 g Zn per hectare. Regardless of the treatment, the highest demand period for B, Cu, Fe, Mn and Zn in both cultivars occurred at the R7 stage (pod formation), i.e. 42 to 55 days after emergence (DAE). The amount of B, Cu, Fe, Mn and Zn exported depended mainly on the level of NPK fertilization used, with values per hectare ranging from 38 to 90 g of B, 12 to 26 g of Cu, 222 to 568 g of Fe 234 to 467 g of Mn, and 40 to 96 g of Zn.

SEQUENTIAL EXTRACTION OF PHOSPHORUS BY MEHLICH-1 AND ION EXCHANGE RESIN FROM B HORIZONS OF FERRIC AND PERFERRIC LATOSOLS (OXISOLS)

In general, Latosols have low levels of available P, however, the influence of the parent material seems to be decisive in defining the pool and predominant form of P in these soils. This study evaluated P availability by extraction with Mehlich-1 (M-1) and Ion Exchange Resin (IER), from samples of B horizons of Ferric and Perferric Latosols developed from different parent materials. To this end, in addition to the physical and chemical characterization of soils, 10 sequential extractions were performed with M-1 and IER from samples of B horizons (depth between 0.8 and 1.0 m). Total contents of Ca, P, Fe, Al, and Ti were determined after digestion with nitric, hydrofluoric and perchloric acids. The effects of sequential P extractions on Fe oxides were also evaluated from the analyses of dithionite-citrate-bicarbonate and ammonium acid oxalate. The high similarity between contents of P accumulated after sequential extractions with M-1 and IER in soils developed on tuffite indicated a predominance of P-Ca. Higher contents of P after a single IER extraction show greater efficiency in P removal from highly weathered soils, as from the Latosols studied here. The P contents also show the high sensitivity of extractant M-1 in highly buffered soils. Furthermore, a single extraction with extractant M-1 or IER is not sufficient to estimate the amount of labile P in these soils.

LOSS OF EXTRACTION CAPACITY OF MEHLICH-1 AND MONOCALCIUM PHOSPHATE AS A VARIABLE OF REMAINING P AND ITS RELATIONSHIP TO CRITICAL LEVELS OF SOIL PHOSPHORUS AND SULFUR

The Mehlich-1 (M-1) extractant and Monocalcium Phosphate in acetic acid (MCPa) have mechanisms for extraction of available P and S in acidity and in ligand exchange, whether of the sulfate of the extractant by the phosphate of the soil, or of the phosphate of the extractant by the sulfate of the soil. In clayey soils, with greater P adsorption capacity, or lower remaining P (Rem-P) value, which corresponds to soils with greater Phosphate Buffer Capacity (PBC), more buffered for acidity, the initially low pH of the extractants increases over their time of contact with the soil in the direction of the pH of the soil; and the sulfate of the M-1 or the phosphate of the MCPa is adsorbed by adsorption sites occupied by these anions or not. This situation makes the extractant lose its extraction capacity, a phenomenon known as loss of extraction capacity or consumption of the extractant, the object of this study. Twenty soil samples were chosen so as to cover the range of Rem-P (0 to 60 mg L-1). Rem-P was used as a measure of the PBC. The P and S contents available from the soil samples through M-1 and MCPa, and the contents of other nutrients and of organic matter were determined. For determination of loss of extraction capacity, after the rest period, the pH and the P and S contents were measured in both the extracts-soils. Although significant, the loss of extraction capacity of the acidity of the M-1 and MCPa extractants with reduction in the Rem-P value did not have a very expressive effect. A “linear plateau” model was observed for the M-1 for discontinuous loss of extraction capacity of the P content in accordance with reduction in the concentration of the Rem-P or increase in the PBC, suggesting that a discontinuous model should also be adopted for interpretation of available P of soils with different Rem-P values. In contrast, a continuous linear response was observed between the P variables in the extract-soil and Rem-P for the MCPa extractor, which shows increasing loss of extraction capacity of this extractor with an increase in the PBC of the soil, indicating the validity of the linear relationship between the available S of the soil and the PBC, estimated by Rem-P, as currently adopted.

Polyethylene glycol and polyvinylpirrolidone effect on bacterial rRNA extraction and hybridization from cells exposed to tannins

In order to detect fluctuations in ruminal microbial populations due to forage tannins using 16S ribosomal RNA (rRNA) probes, recovery of intact rRNA is required. The objective of this work was to evaluate the effect of polyethylene glycol (PEG) and polyvinylpirrolidone (PVP) on extraction of bacterial rRNA, in the presence of tannins from tropical legume forages and other sources, that hybridize with oligonucleotide probes. Ruminococcus albus 8 cells were exposed to 8 g/L tannic acid or 1 g/L condensed tannins extracted from Acacia angustissima, banana (Musa sp.) skin, Desmodium ovalifolium, red grape (Vitis vinifera) skin and Inga edulis, or no tannins. Cells were rinsed with Tris buffer pH 7 containing either 8% PEG or 6% PVP prior to cell lysis. Total RNA samples rinsed with either PEG or PVP migrated through denaturing agarose gels. The 16S rRNA bands successfully hybridized with a R. albus species-specific oligonucleotide probe, regardless of tannin source. The effect of rinsing buffers on the density of 16S rRNA bands, as well as on the hybridization signals was compared. There were significant effects (P<0.01) when the controls were compared to either buffer treatments due to tannin type, buffer used and the interaction of tannin type and buffer. The significant interaction indicates the influence of tannin type on the parameters evaluated.

Influence of rootstocks and pruning times on yield and on nutrient content and extraction in 'Niagara Rosada' grapevine

The objective of this work was to evaluate the influence of rootstocks and pruning times on yield and on nutrient content and extraction by pruned branches and harvested bunches of 'Niagara Rosada' grapevine in subtropical climate. The rootstocks 'IAC 766', 'IAC 572', 'IAC 313', 'IAC 571-6', and '106-8 Mgt' were evaluated. Treatments consisted of a combination between five rootstocks and three pruning times. At pruning, fresh and dry matter mass of branches were evaluated to estimate biomass accumulation. At harvest, yield was estimated by weighing of bunches per plant. Branches and bunches were sampled at pruning and at harvest, respectively, for nutrient content analysis. Nutrient content and dry matter mass of branches and bunches were used to estimate total nutrient extraction. 'Niagara Rosada' grapevine grafted onto the 'IAC 572' rootstock had the highest yield and dry matter mass of bunches, which were significantly different from the ones observed in 'Niagara Rosada'/'IAC 313'. 'Niagara Rosada' grafted onto the 'IAC 572' rootstock extracted the largest quantity of K, P, Mg, S, Cu, and Fe, differing from 'IAC 313' and 'IAC 766' in K and P extraction, and from '106-8 Mgt' in Mg and S extraction. Winter pruning results in higher yield, dry matter accumulation by branches, and total nutrient content and extraction.

Enzymatic technology to improve oil extraction from Caryocar brasiliense camb. (Pequi) Pulp.

The present study aims to compare yield and quality of pequi pulp oil when applying two distinct processes: in the first, pulp drying in a tray dryer at 60ºC was combined with enzymatic treatment and pressing to oil extraction; in the second, a simple process was carried out by combining sun-drying pulp and pressing. In this study, raw pequi fruits were collected in Mato Grosso State, Brazil. The fruits were autoclaved at 121ºC and stored under refrigeration. An enzymatic extract with pectinase and CMCase activities was used for hydrolysis of pequi pulp, prior to oil extraction. The oil extractions were carried out by hydraulic pressing, with or without enzymatic incubation. The oil content in the pequi pulp (45% w/w) and the physicochemical characteristic of the oil was determined according to standard analytical methods. Free fatty acids, peroxide values, iodine and saponification indices were respectively 1.46 mgKOH/g, 2.98 meq/kg, 49.13 and 189.40. The acidity and peroxide values were lower than the obtained values in commercial oil samples, respectively 2.48 mgKOH/g and 5.22 meq/kg. Aqueous extraction has presented lower efficiency and higher oxidation of unsaturated fatty acids. On the other hand, pequi pulp pressing at room temperature has produced better quality oil. However its efficiency is still smaller than the combined enzymatic treatment and pressing process. This combined process promotes cellular wall hydrolysis and pulp viscosity reduction, contributing to at least 20% of oil yield increase by pressing.

COMPARISON OF RNA EXTRACTION METHODS FOR Passiflora edulis SIMS LEAVES

ABSTRACT Functional genomic analyses require intact RNA; however, Passiflora edulis leaves are rich in secondary metabolites that interfere with RNA extraction primarily by promoting oxidative processes and by precipitating with nucleic acids. This study aimed to analyse three RNA extraction methods, Concert™ Plant RNA Reagent (Invitrogen, Carlsbad, CA, USA), TRIzol® Reagent (Invitrogen) and TRIzol® Reagent (Invitrogen)/ice -commercial products specifically designed to extract RNA, and to determine which method is the most effective for extracting RNA from the leaves of passion fruit plants. In contrast to the RNA extracted using the other 2 methods, the RNA extracted using TRIzol® Reagent (Invitrogen) did not have acceptable A260/A280 and A260/A230 ratios and did not have ideal concentrations. Agarose gel electrophoresis showed a strong DNA band for all of the Concert™ method extractions but not for the TRIzol® and TRIzol®/ice methods. The TRIzol® method resulted in smears during electrophoresis. Due to its low levels of DNA contamination, ideal A260/A280 and A260/A230 ratios and superior sample integrity, RNA from the TRIzol®/ice method was used for reverse transcription-polymerase chain reaction (RT-PCR), and the resulting amplicons were highly similar. We conclude that TRIzol®/ice is the preferred method for RNA extraction for P. edulis leaves.

Aplicação de SPME (Solid Phase Micro-Extraction) na análise de águas potáveis de três localidades do estado de São Paulo

The technique of solid phase microextraction (SPME) was used for the extraction of halogenated contaminants of water samples from three cities of the State of São Paulo and the extracts were submitted to gas chromatographic analysis with electron capture detection (GC-ECD). In the samples of water collected at the city of São Paulo the detected level of trihalomethanes (THM) expressed as the sum of chloroform, dibromochloromethane and dichlorobromomethane, were higher than the permissible limit established by the Brazilian regulation. In the samples collected at the two other cities the level of any of the three THM remained below the sensitivity of the ECD.

Solid phase extraction of iron and lead in environmental matrices on amberlite xad-1180/pv

A solid phase extraction procedure using Amberlite XAD-1180/Pyrocatechol violet (PV) chelating resin for the determination of iron and lead ions in various environmental samples was established. The procedure is based on the sorption of lead(II) and iron(III) ions onto the resin at pH 9, followed by elution with 1 mol/L HNO3 and determination by flame atomic absorption spectrometry. The influence of alkaline, earth alkaline and some transition metals, as interferents, are discussed. The recoveries for the spiked analytes were greater than 95%. The detection limits for lead and iron by FAAS were 0.37 µg/L and 0.20 µg/L, respectively. Validation of the method described here was performed by using three certified reference materials (SRM 1515 Apple Leaves, SRM 2711 Montana Soil and NRCC-SLRS-4 Riverine Water). The procedure was successfully applied to natural waters and human hair.

Extraction, purification and biochemical characterization of a peroxidase from Copaifera langsdorffii leaves

The aim of this work is to obtain, purify and characterize biochemically a peroxidase from Copaifera langsdorffii leaves (COP). COP was obtained by acetone precipitation followed by ion-exchange chromatography. Purification yielded 3.5% of peroxidase with the purification factor of 46.86. The COP optimum pH is 6.0 and the temperature is 35 ºC. COP was stable in the pH range of 4.5 to 9.3 and at temperatures below 50.0 ºC. The apparent Michaelis-Menten constants (Km) for guaiacol and H2O2 were 0.04 mM and 0.39 mM respectively. Enzyme turnover was 0.075 s-1 for guaiacol and 0.28 s-1 for hydrogen peroxide. Copaifera langsdorffii leaves showed to be a rich source of active peroxidase (COP) during the whole year. COP could replace HRP, the most used peroxidase, in analytical determinations and treatment of industrial effluents at low cost.