Purification and characterization of polygalacturonase produced by thermophilic Thermoascus aurantiacus CBMAI-756 in submerged fermentation

An extracellular polygalacturonase was isolated from 5-day culture filtrates of Thermoascus aurantiacus CBMAI-756 and purified by gel filtration and ion-exchange chromatography. The enzyme was maximally active at pH 5.5 and 60-65 degrees C. The apparent K (m) with citrus pectin was 1.46 mg/ml and the V (max) was 2433.3 mu mol/min/mg. The apparent molecular weight of the enzyme was 30 kDa. The enzyme was 100% stable at 50 degrees C for 1 h and showed a half-life of 10 min at 60 degrees C. Polygalacturonase was stable at pH 5.0-5.5 and maintained 33% of initial activity at pH 9.0. Metal ions, such as Zn+2, Mn+2, and Hg+2, inhibited 50, 75 and 100% of enzyme activity. The purified polygalacturonase was shown to be an endo/exo-enzyme, releasing mono, di and tri-galacturonic acids within 10 min of hydrolysis.

Redução de sementes do tangor 'Murcote' com a aplicação de biorreguladores durante o florescimento

The present experiment was conducted in a commercial tangor 'Murcote' citrus grove in Pratania, São Paulo State, Brazil and had the objective to evaluate the effects of, 2,4-D (auxin), NAA (auxin), GA(3) (gibberellin) and BA (cytokinin), on the reduction of seed number, without modifications on citrus fruit quality. The treatments sprayed were as follow: control (water); 10 and 20 mg.L-1 of 2,4-D; 100, 150 and 200 mg.L-1 of NAA; 100 and 200 mg.L-1 of GA(3) ; 20 and 40 mg.L-1 of BA. The results showed that none of plant growth regulators influenced fruit quality, without weight reduction, diameter or (0)Brix. In relation to seed number, none of the plant growth regulators were effective on reduction of seed number, however the reduced of viable seed number and total seed number of fruits, specially with the treatment of 100 and 200 mg.L-1 of NAA and 100 mg.L-1 of GA(3), that showed a reduction of 30% of total seed of tangor murcott fruits.

Acerola's pectin methylesterase: studies of heat inactivation

The effect of temperature on the activity of acerola's pectin methylesterase (PME) was studied to determine its heat-inactivation. The acerola's pectin methylesterase (PME; EC: 3.1.1.11) is very stable at 50 degrees C (10% loss of activity in 100 min) and needed 110 min for its inactivation at 98 degrees C. These values are much higher than the ones required for inactivation of the citrus PME, that has been reported as being equal to 1 min at 90 degrees C. Heat-inactivation of PME was shown to be nonlinear, suggesting the presence of fractions of PME with differing heat-stabilities. The times to inactive the enzyme at 98, 102 and 106 degrees C were 110, 10 and 2.17 min, respectively. The Z value (the rise in temperature necessary to observe a ten times faster heat-inactivation) was 4.71 degrees C. (C) 2000 Elsevier B.V. Ltd. All rights reserved.

Effect of lesion age, humidity, and fungicide application on sporulation of Alternaria alternata, the cause of brown spot of tangerine

Alternaria brown spot, caused by Alternaria alternata, causes yield losses and fruit blemishes on many tangerines and their hybrids in most citrus areas of the world where susceptible cultivars are grown. Although the conditions affecting infection and disease severity are known, little information is available on inoculum production on infected tissue. We found that sporulation on leaves began about 10 days after symptoms developed, was abundant from 20 to 40 days, and declined thereafter. Conidial production was far greater on leaf than on fruit or twig lesions. Spore production per unit area of leaf lesion was greater on the more susceptible hybrids, Minneola and Orlando tangelos, than on the less susceptible Murcott tangor. At 74% relative humidity, conidial production on leaf lesions was low, but it was abundant at 85, 92.5, 96, and 100%. Application of Q(o)I or copper fungicides, but not ferbam, suppressed sporulation on leaf lesions for about 14 to 21 days after application. Additional applications did not appear to be more effective than a single spray in reducing inoculum production.

Efeito da lavagem artificial na eficiência de acaricidas-ovicidas no controle de Brevipalpus phoenicis (Geijskes, 1939) (Acari: Tenuipalpidae) em citros

The objective of this research was to test the control of Brevipalpus phoenicis (Geijskes, 1939) by the hexythiazox and quinometionato products in citrus crop with and without adhesive spread and when submitted to artificial rain. The plants were sprayed at 2, 4, 8, 24 and 48 hours. When dried, the fruits were collected (eight/plants) and half of them were washed in laboratory with artificial rainfall on the basis of 30 mm/h during 15 minutes establishing the following treatments: T1 - hexythiazox + washing; T2 - hexythiazox + agral + washing; T3 - quinometionato + washing; T4 - quinometionato + agral + washing; T5 - control + agral + washing; T6 - hexythiazox; T7 - hexythiazox + agral; T8 - quinometionato; T9 - quinometionato + agral; T10 - control + agral. Thus, all the fruits were inoculated with ten females of B. phoenicis and five days later the mites alive were counted and eliminated. Approximately 15 days later the number of larvae alive were also counted. The results obtained allowed the following conclusions: a) the acaricides were efficient to control B. phoenicis; b) the rainfall (washing) did not alter the efficiency; c) the agral did not change the results.

Determination of malic acid in real samples by using enzyme immobilized reactors and amperometric detection

The malate dehydrogenase (MDH) and ascorbate oxidase were immobilized independently, onto silanized controlled porous silica and packed in a tygon tube. The reactors were inserted in the flow system, and the malic acid was determined by measurement of NADH produced by enzymatic reaction. The NADH was reoxidized in a wall jet cell that consisted of spectrographic graphite, Ag/AgCl, KCl(sat), and steel needle as work, reference, and counter electrodes, respectively. The current intensities were measured at 390 mV. The malate calibration curve shows a linear range from 5.0 x 10(-6) to 1.0 x 10(-4) molL(-1), the lifetime was 40 analyses, after that a decrease of 20% on the response is observed. Three different citric juices were analyzed and a good correlation between the proposed method and spectrophotometric commercial kit were obtained.

Effects of plant regulators and potassium nitrate, on Rangpur lime seeds germination

The effects of plant regulators and potassium nitrate were studied on 'Rangpur' lime (Citrus limonia Osbeck) seeds germination. The seeds were removed from ripe fruits, washed, dried and stored at 4 - 5 degrees C, and treated for 24 hours before sowing and the treatments were: KNO3 0,1% and 0,2%; GA(3) 50 ppm, 100 ppm and 250 ppm; GA(4+7)+phenylmethylaminepurine 100 ppm; phenylmethylaminepurine 20 ppm and control. The germination was evaluated begining 13 days after sowing. Best results were obtained with KNO3 0,2%, which proved most beneficial in enhancing percentage and germination rate.

Effect of heated solutions on decay control and residues of imazalil in lemons

Freshly harvested lemons [(Citrus limon (L.) Burm)] were dipped 3 min in water with and without imazalil (IMZ) at 50, 100, and 200 ppm at 50 degrees C and at 1000 ppm IMZ at 20 degrees C. Following treatments fruit were kept at 9 degrees C and 90%-95% relative humidity (RH) for 13 weeks and an additional week at 21 degrees C and ca. 75% RH, to simulate a marketing period (SMP). No decay control was observed with fruit dipped in water at 50 degrees C. In contrast, IMZ treatments provided 90%-96% control of Penicillium rots during cold storage and SMP. Fungi other than Penicillium spp. were also found in all samples as differences among treatments were negligible. IMZ treatment caused some external damage to the fruit (peel browning), and the percentage of damaged fruit was related to the amount of active ingredient (AI) present in it. Dipping in 200 or 1000 ppm IMZ promoted off-flavor development after 10 weeks of storage, and fruit were judged to be unacceptable for consumption after 13 weeks of cold storage. After 1000 ppm IMZ dipping at 20 degrees C, residue concentration in fruit was 8.20 ppm; this value doubled that found in a previous investigation on lemons treated with comparable IMZ levels. Residue concentrations in fruit after treatment at 50 degrees C was strictly related to the amount of fungicide employed. After 13 weeks Al residues in fruit decreased to average ca. 35% of the initial values. During the 1 week SMP, residue levels decreased by a further ca. 25%. It was concluded that it is possible to achieve significant control of decay in lemons during longterm storage by dipping fruit in 50 ppm IMZ mixtures at 50 degrees C. Such treatment should be advised to remarkably reduce potential pollution in the environment due to packinghouse wastewater disposal.

Optimum design of microirrigation systems in sloping lands

When an area to be irrigated has a high slope gradient in the manifold line direction, an option is to use a tapered pipeline to economize on pipe costs and to keep pressure head variations within desired limits. The objective of this paper is to develop a linear optimization model to design a microirrigation system with tapered, downhill manifold lines, minimizing the equivalent annual cost of the hydraulic network and the annual pumping cost, and maximizing the emission uniformity previously established to the subunit. The input data are irrigation system layout, cost of all hydraulic network components, and electricity price. The output data are equivalent annual cost, pipeline diameter in each line of the system, pressure head in each node, and total operating pressure head. To illustrate its capability, the model is applied in a citrus orchard in Sao, Paulo State, Brazil, considering slopes of 3, 6, and 9%. The model proved to be efficient in the design of the irrigation system in terms of the emission uniformity desired.

Use of FIA systems for on-line dilution in multielement determination by ICP-MS

A flow-injection (FI) system to match concentrations was used as an auto-diluter in multielement determination by inductively coupled plasma-mass spectrometry (ICP-MS). The flow system comprised loop-based injection or a timed valve that introduced a variable sample volume info a spray chamber through a standard Meinhard nebulizer of an ICP-MS. Routinely analyzed samples such as water, plant, and steel were selected. The accuracy of multielement determination was checked against water standard reference material from the National Institute of Standards and Technology (1643d), plant standard reference material from the National Bureau of Standards (1572 citrus leaves), and steel standard reference material from the National Bureau of Standards (AISI 4340). The measuring system was calibrated with a multielement solution, yielding a linear plot with good precision [relative standard deviation (RSD) < 3%, n = 12]. The results were in agreement at a 95% confidence level with the certified values for the reference materials and also with those obtained by continuous aspiration and by (FI) with a discrete volume. (C) 1999 John Wiley & Sons, Inc.

Spectrophotometric detection of arsenic using flow-injection hydride generation following sorbent extraction preconcentration

An automated system with a C-18 bonded silica gel packed minicolumn is proposed for spectrophotometric detection of arsenic using flow-injection hydride generation following sorbent extraction preconcentration. Complexes formed between arsenic(III) and ammonium diethyl dithiophosphate (ADDP) are retained on a C-18 sorbent. The eluted As-DDP complexes are merged with a 1.5% (w/v) NaBH4 and the resulting solution is thereafter injected into the hydride generator/gas-liquid separator. The arsine generated is carried out by a stream of N-2 and trapped in an alkaline iodine solution in which the analyte is determined by the arsenomolybdenum blue method. With preconcentration time of 120 s, calibration in the 5.00-50.0 mu g As l(-1) range and sampling rate of about 20 samples h(-1) are achieved, corresponding to 36 mg ADDP plus 36 mg ammonium heptamolybdate plus 7 mg hydrazine sulfate plus 0.7 mg stannous chloride and about 7 mi sample consumed per determination. The detection limit is 0.06 mu g l(-1) and the relative standard deviation (n = 12) for a typical 17.0 mu g As l(-1) sample is ca. 6%. The accuracy was checked for arsenic determination in plant materials from the NIST (1572 citrus leaves; 1573 tomato leaves) and the results were in agreement with the certified values at 95% confidence level. Good recoveries (94-104%) of spiked tap waters, sugars and synthetic mixtures of trivalent and pentavalent arsenic were also found. (C) 1999 Elsevier B.V. B.V. All rights reserved.

Pharmacokinetic Study of Nobiletin and Tangeretin in Rat Serum by High-Performance Liquid Chromatography-Electrospray Ionization-Mass Spectrometry

Nobiletin (NOB) and tangeretin (TAN), two of the main polymethoxylated flavones (PMFs) in citrus, influence a number of key biological pathways in mammalian cells. Although the impacts of NOB and TAN on glucose homeostasis and cholesterol regulation have been investigated in human clinical trials, much information is still lacking about the metabolism and oral bioavailability of these compounds in animals. In this study, NOB and TAN were administered to rats by gavage and intraperitoneal (ip) injection, and the blood serum concentrations of these compounds and their main metabolites were monitored by high-performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS). In addition to the administered compounds, two metabolites of TAN and eight metabolites of NOB were detected and measured over 24 h. With identical oral doses, nearly 10-fold higher absorption of NOB occurred compared to TAN. For both compounds, maximum levels of glucuronidated metabolites occurred in the blood serum at later time points (similar to 5-8 h) compared to the earlier T(max) a values for NOB and TAN. In most cases the glucuronides occurred at substantially higher concentrations than the aglycone metabolites. Low levels of NOB and TAN and their metabolites were detectable in rat blood serum even at 24 h after treatment.

Tolerância de diferentes cultivares de cana-de-açúcar (Saccharum spp.) a herbicidas

This work aimed to evaluate the tolerance of sugarcane cultivars to sulfentrazone, imazapic, isoxaflutole, clomazone and ametryn + trifloxysulfuron-sodium. The experiment was arranged in a randomized block design in a split-plot scheme. The cultivars were allocated to the plots and the herbicides to the sub-plots (five 8.0 m long rows and 1.5 m spacing, with 4 repetitions. The herbicides sulfentrazone (0.8 kg ha(-1)), imazapic (0.147 kg ha(-1)), isoxaflutole (0.1125 kg ha(-1)), clomazone (1.1 kg ha(-1)), ametryn (1.463 kg ha(-1)) + trifloxysulfuron sodium (0.037 kg ha-1) and control were evaluated on 3-yr-old ratoons of the cultivars IACSP94-2094, IACSP94-2101, IACSP93-3046, IACSP94-4004, IAC86-2480 and RB72454 in post emergence. The traits evaluated were: plant toxicity symptoms in the plant leaves; total chlorophyll content and photochemical efficiency (Fv/Fm) at 15, 30 and 60 days after application (DAA); height (cm) at 30 and 270 DAA, and plant stand (stalk m(-1)) at 30 and 180 DAA. Diameter (cm), estimated productivity (t ha(-1)) and quality analysis were evaluated at 270 DAA. The sugarcane cultivars IACSP94-2094, IACSP93-3046, IACSP94-4004, IAC86-2480, RB72454, and IACSP94-2101 especially, were susceptible to clomazone up to 30 DAA, due to leaf chlorosis and lower chlorophyll content, but had no effect on quality characteristics and productivity. The cultivars were also tolerant to other herbicides.

Fruit maturation as a parameter for selection of sweet orange cultivars in Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influence of the cultivation system in the aroma of the volatile compounds and total antioxidant activity of passion fruit

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)