Agricultural Research Service - Pacific West Area

This collaborative study was conducted to determine the total monomeric anthocyanin concentration by the pH differential method, which is a rapid and simple spectrophotometric method based on the anthocyanin structural transformation that occurs with a change in pH (colored at pH 1.0 and colorless at pH 4.5). Eleven collaborators representing commercial laboratories, academic institutions, and government laboratories participated. Seven Youden pair materials representing fruit juices, beverages, natural colorants, and wines were tested. The repeatability relative standard deviation (RSDr) varied from 1.06 to 4.16%. The reproducibility relative standard deviation (RSDR) ranged from 2.69 to 10.12%. The HorRat values were < or = 1.33 for all materials. The Study Director recommends that the method be adopted Official First Action.

Improved molecular disassembly and depolymerization of grain starch to glucose are key to reducing energy use in the bioconversion of glucose to chemicals, ingredients, and fuels. In fuel ethanol production, these biorefining steps use 10-20% of the energy content of the fuel ethanol. The need to minimize energy use and to raise the net yield of energy can be met by replacing high-temperature, liquid-phase, enzymatic digestion with low temperature, solid-phase, enzymatic digestion. Also called cold hydrolysis, the approach is a step toward a "green" method for the production of fuel ethanol. There has been substantial prior and increased recent interest in this approach that is presented in this first review of the subject. We include incentives, developmental research, fundamental factors of raw starch digestion, and novel approaches in enzymology and processing. The discussion draws on resources found in enzymology, engineering, plant physiology, cereal chemistry, and kinetics.

Obesity is a strong risk factor for the development of CVD, hypertension and type 2 diabetes. The overall goal of the present pilot study was to feed strawberries, in the form of freeze-dried powder, to obese subjects to determine whether dietary strawberries beneficially altered lipid profiles and reduced blood markers of inflammation compared with a control intervention. A total of twenty healthy subjects (thirteen females and seven males) aged between 20 and 50 years with a BMI between 30 and 40 kg/m2 completed the present 7-week double-blind, randomised, cross-over trial. Each subject received a prepared diet 7 d/week for 7 weeks consisting of approximately 35 % of energy from fat, 20 % protein, 45 % carbohydrate and 14 g fibre. Blood was collected on days 1 and 8 for baseline information. After the first week, subjects were randomly assigned to the strawberry powder (equivalent to four servings of frozen strawberries) or control (strawberry-flavoured) intervention for 3 weeks. For the remaining 3 weeks, subjects crossed over to the opposite intervention. Blood was collected again at the end of weeks 3, 4, 6 and 7. A comprehensive chemistry panel, lipid profile analyses and measurement of inflammatory mediators were performed for each blood draw. A 3-week dietary intervention with strawberry powder reduced plasma concentrations of cholesterol and small HDL-cholesterol particles, and increased LDL particle size in obese subjects (P < 0·05). Dietary strawberry powder reduced risk factors for CVD, stroke and diabetes in obese volunteers, suggesting a potential role for strawberries as a dietary means to decrease obesity-related disease.

To understand the mechanisms responsible for aluminum (Al) toxicity and tolerance in plants, an expressed sequence tag (EST) approach was used to analyze changes in gene expression in roots of rye (Secale cereale L. cv Blanco) under Al stress. Two cDNA libraries were constructed (Al stressed and unstressed), and a total of 1,194 and 774 ESTs were generated, respectively. The putative proteins encoded by these cDNAs were uncovered by Basic Local Alignment Search Tool searches, and those ESTs showing similarity to proteins of known function were classified according to 13 different functional categories. A total of 671 known function genes were used to analyze the gene expression patterns in rye cv Blanco root tips under Al stress. Many of the previously identified Al-responsive genes showed expression differences between the libraries within 6 h of Al stress. Certain genes were selected, and their expression profiles were studied during a 48-h period using northern analysis. A total of 13 novel genes involved in cell elongation and division (tonoplast aquaporin and ubiquitin-like protein SMT3), oxidative stress (glutathione peroxidase, glucose-6-phosphate-dehydrogenase, and ascorbate peroxidase), iron metabolism (iron deficiency-specific proteins IDS3a, IDS3b, and IDS1; S-adenosyl methionine synthase; and methionine synthase), and other cellular mechanisms (pathogenesis-related protein 1.2, heme oxygenase, and epoxide hydrolase) were demonstrated to be regulated by Al stress. These genes provide new insights about the response of Al-tolerant plants to toxic levels of Al.

Abstract Atmospheric concentrations of carbon dioxide (CO2) may double during the next century, causing changes in the Earth's climate. Warming of up to 4°C, slight cooling, and 10% changes in precipitation have been projected. Researchers have studied the possible impacts these changes may have on various aspects of the hydrological cycle, but little emphasis has been placed on snow accumulation and melt. In this study, the effects of climatic change on streamflow from a snowmelt-dominated basin in southwestern Montana, USA, are investigated. The National Weather Service River Forecast System model (NWSRFS) was first calibrated using data for the 1973–1984 period. Daily temperature and precipitation values were then changed, and the model ran again to assess the effects on snowpack and streamflow of some possible climatic changes. Results indicate that streamflow may vary by from −22 to +45% depending on the combination of climatic changes imposed.

The gluten protein found in a variety of cereal grains is a food allergen that can elicit a spectrum of immuno-inflammatory responses in people. Consumer awareness has prompted changes in food labeling requirements, expanded gluten-free food product availability and increased demand for effective gluten testing methodologies. To meet the challenges associated with gluten testing from diverse and complex foods we developed a lateral flow immunoassay (LFIA) using a pair of novel gliadin monoclonal antibodies (MAbs). Using a visual gold reporter, we show sensitive gluten detection (150 ng/mL) from complex food substrates using a fast (<5 min) and easy testing methodology. In this report we characterize the binding properties of a cohort of newly generated gliadin monoclonal antibodies suitable for gluten detection using multiple assay formats and introduce a novel plug-n-play test strip platform with integrated test components in a single-use format.

Abstract The output from a pesticide screening model indicated that chemicals with low Henry's Law constants ( K H ) will tend to accumulate at the soil surface when water is evaporating, resulting in an increased volatilization with time. Volatilization losses and surface distributions of two pesticides with widely differing K H , were measured to test the predictions of the screening model, particularly with respect to the importance of K H in controlling relative volatilization and vapor behavior. Volatilization of prometon (2,4‐bis(isopropylamino)‐6‐methoxy‐ s ‐triazine], a Category III compound with low K H , increased with time with water evaporating and prometon accumulated at the soil surface. Volatilization of lindane (gamma isomer of 1,2,3,4,5,6‐hexachloro‐cyclohexane), a Category I compound with high K H , decreased with time with or without water evaporating and did not accumulate at the surface with water evaporating. The experimental data confirmed the model predictions that volatilization of chemicals with low K H is controlled by the air‐boundary layer above the soil surface, whereas control of volatilization of Category I chemicals with high K H is within the soil. Accumulation and increased volatilization of compounds with low K H are controlled by boundary layer thickness and water evaporation rate as well as K H . The phenomena of organic chemicals with low K H accumulating at the soil surface following convective movement in evaporating water could greatly enhance their volatilization and increase their availability for photolysis and runoff into surface waters. Sufficient accumulation can occur that changes in adsorption coefficients with concentration must be taken into account in modeling their volatilization.

This review presents an update on the physical, chemical, and biological properties of food allergens in plant sources, focusing on the few protein families that contribute to multiple food allergens from different species and protein families recently found to contain food allergens. The structures and structural components of the food allergens in the allergen families may provide further directions for discovering new food allergens. Answers as to what makes some food proteins allergens are still elusive. Factors to be considered in mitigating food allergens include the abundance of the protein in a food, the property of short stretches of the sequence of the protein that may constitute linear IgE binding epitopes, the structural properties of the protein, its stability to heat and digestion, the food matrix the protein is in, and the antimicrobial activity to the microbial flora of the human gastrointestinal tract. Additionally, recent data suggest that widely used techniques for mapping linear IgE binding epitopes need to be improved by incorporating positive controls, and methodologies for mapping conformational IgE binding epitopes need to be developed.

Almond (Prunus dulcis) is not only widely used as a human food as a result of its flavor, nutrients, and health benefits, but it is also one of the most likely tree nuts to trigger allergies. Almond allergens, however, have not been studied as extensively as those of peanuts and other selected tree nuts. This review provides an update of the molecular properties of almond allergens to clarify some confusion about the identities of almond allergens and our perspective on characterizing putative almond allergens. At present, the following almond allergens have been designated by the World Health Organization/International Union of Immunological Societies Allergen Nomenclature Sub-Committee: Pru du 3 (a non-specific lipid transfer protein 1, nsLTP1), Pru du 4 (a profilin), Pru du 5 (60S acidic ribosomal protein 2), Pru du 6 (an 11S legumin known as prunin) and Pru du 8 (an antimicrobial protein with cC3C repeats). Besides, almond vicilin and almond γ-conglutin have been identified as food allergens, although further characterization of these allergens is still of interest. In addition, almond 2S albumin was reported as a food allergen as a result of the misidentification of Pru du 8. Two more almond proteins have been called allergens based on their sequence homology with known food allergens and their 'membership' in relevant protein families that contain allergens in many species. These include the pathogenesis related-10 protein (referred to as Pru du 1) and the thaumatin-like protein (referred to as Pru du 2). Almonds thus have five known food allergens and five more likely ones that need to be investigated further. Published 2020. This article is a U.S. Government work and is in the public domain in the USA.

Abstract Soybean [Glycine max(L.) Merrill] is moderately salt tolerant, but the method of irrigation used for crop production under saline conditions may influence the uptake and distribution of potentially toxic salts. This field study was conducted to determine the effects of application of saline waters by different methods, namely, drip and above‐canopy sprinkler irrigation, on the ion relations of soybean cultivar “Manokin”. Salinity was imposed by adding NaCl and CaCl2 (1:1 by weight) to nonsaline irrigation waters. Saline treatments with electrical conductivity (EC i ) of 4 dS m−1 were compared with nonsaline controls (EC i = 0.5 dS m−1). Ion concentrations in leaves, stems, roots, and when present, pods were determined at four stages of growth: vegetative, flowering, podding, and grain filling. Both Na+ and Cl− were excluded from the Manokin leaves and stems when plants were drip‐irrigated and the uptake of these ions occurred solely via the root pathway. However, when saline water was applied by sprinkling, the ions entered leaves by both foliar absorption and root uptake and their concentrations in the leaves were about 9‐fold higher than in those under saline drip irrigation. Regardless of treatment, leaf‐K was highest during the vegetative stage, then decreased with plant age as K+ was mobilized to meet nutrient demands of the developing reproductive structures.

ABSTRACT Wheat gluten reacts with phosphoric acid in the presence of urea to produce natural superabsorbent gels. Fourier Transform Infra‐red (FT‐IR) spectroscopy and two‐dimensional gel electrophoresis (2DE) reveal chemical changes from the reaction. Temperatures above 120°C and dry conditions create the opportunity for reaction. FT‐IR analyses confirm the formation of esters, carbamates, and phosphoramides on the gluten samples. 2DE protein composition topographies indicate a shift in the isoelectric point (p I ) to lower values along with extensive inter‐protein linkages. A free swelling capacity (FSC) in excess of 85× the mass of the converted gluten is obtainable using a conservative vacuum‐assisted method to recover and quantify the properties of the wet gel. Other methods produce FSC values nearly twice as high. FSC for acid‐treated gluten is lower for solutions containing solutes than the FSC for deionized water. Native gluten produces FSC values that are about 2% of those for treated gluten, but these values are less sensitive to the presence of ionic solutes and increase slightly in the presence of aqueous ethanol up to a mole fraction of 0.25.. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131 , 39440.

Restoration in dryland ecosystems is hindered by low establishment of seeded species. As such, evaluations of current seeding methods are critical to understanding limitations and barriers to seeding success. Drill seeding is perceived as an optimal seeding strategy in many dryland ecosystems, but broadcast seeding is more commonly used as a seeding method due to physical and logistical constraints. For example, broadcast seeding may be conducted by aerial drops where other methods are limited by topography or obstructive features in the landscape. Few studies have quantified the differences between drill and broadcast seeding through space and time. We compare 2‐year recruitment of emergent Pseudoroegneria spicata (bluebunch wheatgrass) seedlings in the sagebrush steppe biome for drill versus broadcast seeding methods across three seeding years, three landscape aspects and two soil types using a 95% confidence interval approach to avoid the penalty of multiplicity. We found drill seeding had 2.7 times greater recruitment of seedlings after 2 years compared with broadcast seeding. However, differences were highly subject to seeding year, aspect and soil type, likely because of soil moisture and temperature variations. Drill seeding had an advantage on clay soils with flat and north aspects (10.1 and 4.6 times greater for drill than broadcast seeding, respectively). In most conditions, drill seeding had greater recruitment than broadcast seeding, though in 2014 on south aspects broadcast seeding had 2.7 times greater recruitment than drill seeding. The results of this study demonstrate a need for restoration plans that account for spatiotemporal variation in seeding success.

Obesity increases the risk of developing bacterial and viral infections compared with normal weight. In a 7-week double-blind, randomised, cross-over trial, twenty obese volunteers (BMI between 30 and 40 kg/m²) were fed freeze-dried strawberry powder or strawberry-flavoured placebo preparations to determine the effects of dietary strawberries on immune function. Blood was collected at six time points during the study and peripheral blood mononuclear cells (PBMC) were isolated at each time point and activated with CD3 plus CD28 antibodies (T-lymphocyte activation) or lipopolysaccharide (LPS, monocyte activation). Interferon-γ, TNF-α, IL-4 and IL-10 were measured in supernatants from the activated T cells. Supernatants from the activated monocytes were analysed for the production of TNF-α, IL-1β, IL-6 and IL-8. PBMC were pre-stained with PKH (Paul Karl Horan) dye and activated with CD3 plus CD28 antibodies to determine the proliferative responses of CD4⁺ and CD8⁺ T-lymphocytes by flow cytometry. To detect global changes in gene expression, microarray analysis was performed on LPS- and vehicle-treated PBMC from two subjects before and after the strawberry intervention. No difference was observed for the production of T-cell cytokines between the intervention groups. The production of TNF-α was increased in the supernatants from LPS-activated PBMC in the group consuming strawberries compared with the placebo. A modest increase in the proliferation of the CD8⁺ T-lymphocyte population was observed at 24 h post-activation. These data suggest that dietary strawberries may increase the immunological response of T-lymphocytes and monocytes in obese people who are at greater risk for developing infections.

Biochemical characterizations of food allergens are required for understanding the allergenicity of food allergens. Such studies require a relatively large amount of highly purified allergens. The level of Pru du 4 in almond is low, and its expression in a soluble form in Escherichia coli required an expression tag. An MBP tag was used to enhance its expression and solubility. Sumo was used for the first time as a peptidase recognition site. The expression tag was removed with a sumo protease, and the resulting wild-type Pru du 4 was purified chromatographically. The stability of the allergen was investigated with chemical denaturation. The Gibbs free energy of Pru du 4 folding-unfolding transition was determined to be 5.4 ± 0.7 kcal/mol.

An N-glycosylated 60-kDa PV-1 protein that binds heparin was detected in mouse lung from a single mRNA transcript. In the absence of disulfide bond reduction PV-1 is detected as a dimer or large molecular weight oligomer. In the lung of Cav-1, but not Cav-2, null mice the amount of PV-1 protein is diminished, with no detectable change in mRNA level. PV-1 does not fractionate with caveolae on a sucrose density gradient, but the Cav-1 protein is detected in fractions following immunoprecipitation with PV-1 antibodies. Both PV-1 and Cav-1 localize to alveolar endothelial cells, but PV-1 is concentrated at the abluminal and Cav-1 at the luminal cell surface with minimal colocalization. In the Cav-1 null lungs, PV-1 is nearly undetectable in endothelial cells, but remains unchanged in pneumocytes and bronchial epithelial cells. Injection of a VEGF-R2 inhibitor increased PV-1 protein in lung of Cav-1 null, but not Cav-2 or wild-type mice. These data indicate that the PV-1 protein is negatively regulated in pulmonary endothelial cells by VEGF-R2 signaling.

on the rumen microbiota. For each dosage, 3 animals were randomly assigned into groups and fed the same basal total mixed ration diet supplemented with monensin, at the respective dose. After 20 days, rumen samples were taken and the effect on the microbiota was examined by 16S rRNA gene sequence analysis and qPCR. At the lowest dose no significant change in 16S rRNA gene sequences associated with any bacterial phyla was observed; however, at the medium and high dosages, we observed significant reductions in sequences associated with gram-positive bacteria and significant increases in those associated with gram-negative bacteria that were dosage dependent. All dosages reduced the levels of sequences associated with methanogenic archaea in the rumen, with the medium dosage showing the largest decline. No significant difference was observed for the 18S rRNA gene sequences associated with protozoa in any of the libraries. Our results indicate that with this diet the medium dosage of monensin was most efficacious for the reduction in methanogenic archaea in the rumen of lactating dairy.

Studies on genetic diversity and recombination in bacterial pathogens are providing a better understanding of the mechanisms shaping bacterial diversity, which can affect disease control. Xanthomonas campestris pv. vitians, causal agent of bacterial leaf spot of lettuce, is a threat to the worldwide lettuce industry. We examined the genetic variation within a sample of 83 strains from California, Florida, and Ohio using multilocus sequence typing of six housekeeping genes, totaling 2.7 kb. Additionally, polymorphism in two virulence-related genes, hrpB2 and a putative glycosyl hydrolase, were examined. Based on housekeeping genes, we found three genetic groups of strains that were all able to induce the disease. These included strains collected from weeds and irrigation water that had haplotypes identical to strains from diseased lettuce. High linkage disequilibrium across the sequenced loci indicates that the pathogen is predominantly clonal but recombination has contributed to the observed sequence variation. Although there was significant genetic variation in X. campestris pv. vitians within and among sampled states, identical haplotypes were observed across all three states. This finding suggests that seedborne inoculum may contribute to the diversity of X. campestris pv. vitians in the United States. Knowledge of the genetic structure of the pathogen may be used for developing resistant lettuce varieties.

Core Ideas Alleys have greater soil C and N pools than vine rows. Soil N 2 O emissions increase in response to precipitation, fertigation, and irrigation. Path analysis revealed that vineyard zone and soil texture influence N 2 O emissions. Path analysis revealed that GWC influences N 2 O emissions. To evaluate the effect of precipitation events, management practices, and soil type in wine grape ( Vitis vinifera L. ssp. vinifera ) vineyard systems and provide data for greenhouse gas (GHG) emissions calculators, we monitored nine vineyards in the Lodi wine grape district, California, from April 2011 to December 2012. These commercial vineyards exist on three soil series (Redding, San Joaquin, and Tokay), representing a spectrum of soil textures and degrees of soil development. We hypothesized that soil characteristics would be a dominating factor affecting GHG fluxes, but the magnitude of fluxes would be influenced by precipitation and management events. We measured N 2 O fluxes, soil NO 3 –N and NH 4 –N, and gravimetric water content (GWC) from vine and intervine (alleys) rows bimonthly (April–October) and monthly (November–March). Monthly, we collected soil samples for dissolved organic C (DOC) and dissolved organic N (DON) determination. Path analysis revealed that the effects of soil type and vineyard zone on N 2 O emissions were influenced by soil texture (i.e., gravel and clay contents) but that this effect was mediated by GWC through soil temperature and soil inorganic N content. Management practices such as irrigation, fertigation, cover cropping, and tillage affected differences between vine rows and alleys for soil inorganic pools, DOC, and DON from June to October 2012. This 20‐mo study indicated that precipitation events strongly influenced N 2 O fluxes.

Almond is one of the tree nuts listed by U.S. FDA as a food allergen source. A food allergen identified with patient sera has been debated to be the 2S albumin or the 7S vicilin. However, neither of these proteins has been defined as a food allergen. The purpose of this study was to clone, express, and purify almond vicilin and test whether it is a food allergen. Western blot experiment was performed with 18 individual sera from patients with double-blind, placebo-controlled clinical almond allergy. The results showed that 44% of the sera contained IgE antibodies that recognized the recombinant almond vicilin, indicating that it is an almond allergen. Identifying this and additional almond allergens will facilitate the understanding of the allergenicity of seed proteins in tree nuts and their cross-reactivity.

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