Diversity in White Lupin (<Emphasis Type="Italic">Lupinus albus</Emphasis> L.) Landraces from Northwest Iberianplateau

All the accessions (35) of white lupin (Lupinus albus L.) landraces collected from northwest Iberian plateau and maintained at the Spanish germplasm collection (CRF of INIA), were studied with the general objective of assessing the biodiversity of these landraces and to ascertain their value as genetic resources for the development of germplasm adapted to the areas where they were collected with long and chilly winters. The characterization study comprised 51 qualitative and 50 quantitative characters. Quantitative parameters were analyzed by Principal Components Analysis (PCA). The 2-dimensional plot (49.3% of cumulative variability) formed a main group of accessions and 4 outliers (#1, #9, #27 and #28) separated in the first PC. The characters with a higher contribution to the first PC were inflorescence length, leaf petiole length and leaf central foliole area. The presence of alkaloids, the percentage of plants killed by soil borne fungal diseases, the phenology, the yield per plant and yield components were also assessed. Twenty six accessions (the main group in the PCA plot) showed high homogeneity, with the following highlights: bitterness, indeterminate growth habit, early ripeness and spring sowing with the exception of #18, in which seeding date is unknown and it had a later phenology. By contrast accessions #9 and #27 were bitter of autumn sowing with determinate growing habit and they had the longest values of primary inflorescence length and a later phenology when they were sown in spring. Average values for yield and yield components showed a broad range of variation between accessions. The main yield component was the number of pods per plant. Accession #22 had the highest mean seed mass, although this had not significant influence in yield. Accession #17 showed the highest yield and #1 the lowest. This last also had the lowest values of inflorescence length, leaf petiole length and leaf central foliole area. No significant differences of resistance to soil-borne fungi were found between accessions. The year had a significant effect in the phenology, yield and yield components except for number of seeds per pod. The studied material might be of interest for the development of spring sowing germplasm adapted to north Iberian peninsula.     

Enzymatic dehydrogenative polymerization of urushiols in fresh exudates from the lacquer tree, Rhus vernicifera DC

Fresh exudates from the lacquer tree, Rhus vernicifera DC, were extracted with acetone and the solution was chromatographed to isolate monomer, dimer, trimer, and oligomer fractions of urushiols. Constituents of the monomeric and dimeric fractions were then identified by two-dimensional (2D) 1H-13C heteronuclear multiple quantum coherence (HMQC) and heteronuclear multiple bond coherence (HMBC) NMR spectroscopic techniques. The results showed that the monomeric fraction contained 3-[8'Z,11'E,13'Z-pentadecatrienyl]catechol (1), 3-[8'Z,11'Z,14'-pentadecatrienyl]catechol (2), and 3-pentadecanyl]catechol (3), which was verified by HPLC analysis. The dimeric fraction contained 8'-(3' ',4' '-dihydroxy-5' '-alkenyl)phenyl-3-[9'E,11'E,13'Z-pentadecatrienyl]catechol (4), 14'-(3' ',4' '-dihydroxy-5' '-alkenyl)phenyl-3-[8'Z,10'E,12'E-pentadecatrienyl]catechol (5), 2-hydroxyl-3- or -6-alkenylphenyl ethyl ether (6), 14'-(3' ',4' '-dihydroxy-2' '-alkenyl)phenyl-3-[8'Z,10'E,12'E-pentadeca-trienyl]catechol (7), 15'-(2' '-hydroxy-3' '- or -6' '-alkenyl)phenyloxy-3-[8'Z,11'Z,13'E)-pentadecatrienyl]catechol (8), 14'-(2' ',3' '-dihydroxy-4' '-alkenyl)phenyl-3-[8'Z,10'E,12'E-pentadecantrienyl]catechol (9), 1,1',2,2'-tetrahydroxy-6,6'-dialkenyl-4,3'-biphenyl (10), 1,1',2,2'-tetrahydroxy-6,6'-dialkenyl-4,4'-biphenyl (11), 1,1',2,2'-tetrahydroxy-6,6'-dialkenyl-5,4'-biphenyl (12), and 1,2,1'-trihydroxy-6,6'-dialkenyldibenzofuran (13) as constituents. In addition, dimeric ethers and peroxides, such as compounds 14 and 15, were produced by autoxidation of monomeric urushiols in atmospheric air. The possible reaction mechanisms for the dehydrogenative polymerization of urushiols by Rhus laccase present in the fresh raw exudates under the atmospheric oxygen are discussed on the basis of structures identified. This is of primary importance because the use of the urushi exudates as coating materials does not involve organic solvents and is an environmentally friendly process.     

A calorimetric study of thermal denaturation of recombinant human lactoferrin from rice

Thermal denaturation of recombinant human lactoferrin from transgenic rice with different degrees of iron saturation has been studied by differential scanning calorimetry. The maximum temperature, enthalpy change, and activation energy of denaturation were higher when recombinant lactoferrin was more saturated with iron, indicating an increase in the stability of the protein structure. Maximum temperature and activation energy values for apo- and holo-lactoferrins were practically identical to those reported for the same forms of lactoferrin from human milk, which indicates a similar thermal stability. However, the value of enthalpy change for denaturation of the recombinant lactoferrin was 2.5-3-fold lower than that found for the human milk protein. This finding may reflect the influence that the different glycosylation pattern may have in the relationship between lactoferrin domains. Denaturation of recombinant lactoferrin in milk was compared with denaturation in phosphate buffer, and results indicated that the protein was more heat-sensitive when treated in milk than in buffer.     

Broccoli processing wastes as a source of peroxidase

A peroxidase isozyme (BP) was purified to homogeneity from broccoli stems ( Brassica oleraceae var. maraton) discarded from industrial processing wastes. BP specific activity was 1216 ABTS [2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)] units/mg, representing 466-fold that of crude extract. BP is a monomeric glycoprotein containing 16% carbohydrates, with a molecular mass of 49 kDa and an isoelectric point close to 4.2. From kinetic data it showed a two-substrate ping-pong mechanism, and the catalytic efficiency measured as the rate-limiting step of free BP regeneration was 3.4 x 10(6) M(-1) s(-1). The ABTS K m value was 0.2 mM, which was about 20 times lower than that reported for acidic commercial horseradish peroxidase (HRP). Assessment of BP secondary structure showed 30% helical character, similar to HRP and cytochrome c peroxidase. BP lost only 25% activity after 10 min of heating at 55 degrees C and pH 6; it was stable in the pH range from 4 to 9 and showed an optimum pH of 4.6 using ABTS as substrate. BP was active on substrates normally involved in lignin biosynthesis, such as caffeic and ferulic acids, and also displayed good catechol oxidation activity in the presence of hydrogen peroxide. Reverse micellar extraction was successfully used as potential large-scale prepurification of broccoli peroxidase, achieving a purification factor of 7, with 60% activity yield. Stems from the broccoli processing industry have a high potential as an alternative for peroxidase purification.     

Enhanced Bacillus thuringiensis Production from Sewage Sludge with Alkaline and Ultrasonic Pretreatments

Sewage sludge is a cost-effective media for the production of Bacillus thuringiensis (Bt) based biopesticides. To enhance the entomotoxicity of the fermentation broth, pretreatments of sewage sludge by alkali and ultrasonic were applied in this study. Effects of alkaline and ultrasonic pretreatments on the soluble COD (SCOD) and total COD (TCOD) were evaluated by altering the alkali addition dose and the ultrasonic specific energy. Suitable pretreatment conditions were optimized with 5 g l^?1 sodium hydroxide (NaOH) for alkaline treatment and 1.2?×?10⁵ kJ kg^?1 of total solid for ultrasonic treatment. Fermentations of raw and pretreated sludge for biopesticides were carried out in a bench scale fermentor. Results revealed that both pretreatments were effective for Bt growth and metabolism. Higher viable cells (VC) and viable spores (VS) counts, δ-endotoxin yields and entomotoxicity were achieved in the pretreated sludge. The enhancement was attributed to more available nutrients and better oxygen transfer. Moreover, ultrasonic pretreated sludge was superior to alkaline pretreated sludge for δ-endotoxin production and entomotoxicity owing to its higher soluble C/N ratio and finer particles.     

Enzyme activities and microbial community structure in semiarid agricultural soils

This study investigated the effect of management on -glucosidase, -glucosaminidase, alkaline phosphatase, and arylsulfatase activities and the microbial community structure in semiarid soils from West Texas, USA. Surface samples (0–5 cm) were taken from a fine sandy loam, sandy clay loam, and loam that were under continuous cotton ( Gossypium hirsutum L.) or in cotton rotated with peanut ( Arachis hypogaea L.), sorghum ( Sorghum bicolor L.), rye ( Secale cereale) or wheat ( Triticum aestivum L.), and had different water management (irrigated or dryland), and tillage (conservation or conventional). The enzyme activities were higher in the loam and sandy clay loam than in the fine sandy loam. Soil pH was not affected by management, but the soil organic C and total N contents were generally affected by the different crop rotations and tillage practices studied. The trends of the enzyme activities as affected by management depended on the soil, but in general crop rotations and conservation tillage increased the enzyme activities in comparison to continuous cotton and conventional tillage. The soil enzyme activities were significantly correlated with the soil organic C ( r -values up to 0.90, P< 0.001), and were correlated among each other ( r -values up to 0.90, P <0.001). There were differences in the fatty acid methyl ester profiles between the fine sandy loam and the sandy clay loam and loam, and they reflected the differences in the enzyme activities found among the soils. For example, a 15:0 ranged from 1.61±0.25% in cotton-peanut/irrigated/no-till in the fine sandy loam to 3.86±0.48% in cotton-sorghum/dryland/conservation tillage in the sandy clay loam. There were no differences due to management within the same soil.Trade names and company names are included for the benefit of the reader and do not infer any endorsement or preferential treatment of the product by USDA-ARS     

Organophosphorus pesticide residues in Mexican commercial pasteurized milk

A study was conducted to measure residues of 13 organophosphorus (OP) pesticides, widely used as dairy cattle ectoparasiticides or in crops used for animal feed, in homogenized and pasteurized Mexican milk samples. Four different milk brands with high distribution were collected biweekly during a 12 month period (n = 96) in supermarkets. OP pesticide residues were measured by gas chromatography with a flame photometric detector. Approximately 39.6% of the samples contained detectable levels of OP pesticide residues. Eight samples contained residues exceeding established maximum residue limits (MRL), and the OP pesticides present in these samples were dichlorvos (five samples), phorate, chlorpyrifos, and chlorfenvinphos (one sample, respectively). Average residues of 13 OP pesticides measured were below established MRLs ranging between 0.0051 and 0.0203 ppm.     

Varietal tolerance to Zinc deficiency in wheat and barley grown in chelatorbuffered nutrient solution and its effect on uptake of Cu,Fe, and Mn

Tolerance to zinc (Zn) deficiency was examined for three wheat (Triticum aestivum L.) and three barley (Hordeum vulgare L.) varieties grown in chelator‐buffered nutrient solution. Four indices were chosen to characterize tolerance to Zn deficiency: (1) relative shoot weight at low compared to high Zn supply (“Zn efficiency index”), (2) relative shoot to root ratio at low compared to high Zn supply, (3) total shoot uptake of Zn under deficient conditions, and (4) shoot dry weight under deficient conditions. Barley and wheat exhibited different tolerance to Zn deficiency, with barley being consistently more tolerant than wheat as assessed by all four indices. The tolerance to Zn deficiency in the barley varieties was in the order Thule=Tyra>Kinnan, and that of wheat in the order Bastian=Avle>Vinjett. The less tolerant varieties of both species accumulated more P in the shoots than the more tolerant varieties. For all varieties, the concentrations of Mn, Fe, Cu, and P in shoot tissue were negatively correlated with Zn supply. This antagonism was more pronounced for Mn and P than for Cu and Fe. Accumulation of Cu in barley roots was extremely high under Zn‐deficient conditions, an effect not so clearly indicated in wheat.     

Usefulness of amino acid composition to discriminate between honeydew and floral honeys. Application to honeys from a small geographic area

With the aim of finding methods that could constitute a solid alternative to melissopalynological and physicochemical analyses to determine the botanical origin (floral or honeydew) of honeys, the free amino acid content of 46 honey samples has been determined. The honeys were collected in a small geographic area of approximately 2000 km(2) in central Spain. Twenty-seven honey samples were classified as floral and 19 as honeydew according to their palynological and physicochemical analyses. The resulting data have been subjected to different multivariant analysis techniques. One hundred percent of honey samples have been correctly classified into either the floral or the honeydew groups, according to their content in glutamic acid and tryptophan. It is concluded that free amino acids are good indicators of the botanical origin of honeys, saving time compared with more tedious analyses.