Effects of genetically modified plants on microbial communities and processes in soil

The development and use of genetically modified plants (GMPs) has been a topic of considerable public debate in recent years. GMPs hold great promise for improving agricultural output, but the potential for unwanted effects of GMP use is still not fully understood. The majority of studies addressing potential risks of GMP cultivation have addressed only aboveground effects. However, recent methodological advances in soil microbial ecology have allowed research focus to move underground to try to gain knowledge of GMP-driven effects on the microbial communities and processes in soil that are essential to key terrestrial ecosystem functions. This review gives an overview of the research performed to date on this timely topic, highlighting a number of case studies. Although such research has advanced our understanding of this topic, a number of knowledge gaps still prevent full interpretation of results, as highlighted by the failure of most studies to assign a definitively negative, positive or neutral effect to GMP introduction. Based upon our accumulating, yet incomplete, understanding of soil microbes and processes, we propose a synthesis for the case-by-case study of GMP effects, incorporating assessment of the potential plant/ecosystem interactions, accessible and relevant indicators, and tests for unforeseen effects.     

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     

Analysis of strigolactones,germination stimulants for striga and orobanche,by high-performance liquid chromatography/tandem mass spectrometry

A simple and rapid analytical method for strigolactones, germination stimulants for the root parasitic weeds witchweed (Striga spp.) and broomrape (Orobanche spp.), has been developed using high-performance liquid chromatography connected to tandem mass spectrometry (LC/MS/MS). The natural strigolactones (strigol, sorgolactone, orobanchol, and alectrol) were clearly separated and identified by LC/MS/MS. As low as 0.1 pg/microL of strigol and 0.5 pg/microL of sorgolactone could be quantified, whereas 1 pg/microL was needed for the quantification of orobanchol (S/N > 10). Using this method, it was found that red clover produces orobanchol and alectrol but not strigol. The roots of red clover seedlings were found to produce 13, 70, 58, and 65 pg of orobanchol/plant 1, 2, 3, and 4 weeks after germination, respectively.     

Effect of germination on the carbohydrate composition of the dietary fiber of peas (Pisum sativum L.)

The effect of different conditions of pea germination on dietary fiber (DF) composition was studied. Insoluble dietary fiber (IDF) and soluble dietary fiber (SDF) were subjected to acid hydrolysis, and the resultant neutral sugars, uronic acids, and Klason lignin were quantified. Germinated peas exhibited significantly higher contents of total dietary fiber (TDF) than the raw sample, due to the increases of both DF fractions. Under darkness conditions, germination exhibited the highest contents of IDF and SDF. Decreasing IDF/SDF ratios showed that the carbohydrate changes did not take place to the same extent during germination, the SDF fraction being the most affected. The detailed chemical composition of fiber fractions reveals increases of cellulose in the IDF of germinated samples, whereas SDF exhibits a decrease of pectic polysaccharides and also increases of polysaccharides rich in glucose and mannose. The DF results were corroborated by a comparative examination of the cell wall carbohydrate composition.     

Sinapate dehydrodimers and sinapate-ferulate heterodimers in cereal dietary fiber

Two 8-8-coupled sinapic acid dehydrodimers and at least three sinapate-ferulate heterodimers have been identified as saponification products from different insoluble and soluble cereal grain dietary fibers. The two 8-8-disinapates were authenticated by comparison of their GC retention times and mass spectra with authentic dehydrodimers synthesized from methyl or ethyl sinapate using two different single-electron metal oxidant systems. The highest amounts (481 microg/g) were found in wild rice insoluble dietary fiber. Model reactions showed that it is unlikely that the dehydrodisinapates detected are artifacts formed from free sinapic acid during the saponification procedure. The dehydrodisinapates presumably derive from radical coupling of sinapate-polymer esters in the cell wall; the radical coupling origin is further confirmed by finding 8-8 and 8-5 (and possibly 8-O-4) sinapate-ferulate cross-products. Sinapates therefore appear to have an analogous role to ferulates in cross-linking polysaccharides in cereal grains and presumably grass cell walls in general.     

Lipid oxidation in corn oil-in-water emulsions stabilized by casein,whey protein isolate,and soy protein isolate

Proteins can be used to produce cationic oil-in-water emulsion droplets at pH 3.0 that have high oxidative stability. This research investigated differences in the physical properties and oxidative stability of corn oil-in-water emulsions stabilized by casein, whey protein isolate (WPI), or soy protein isolate (SPI) at pH 3.0. Emulsions were prepared with 5% corn oil and 0.2-1.5% protein. Physically stable, monomodal emulsions were prepared with 1.5% casein, 1.0 or 1.5% SPI, and > or =0.5% WPI. The oxidative stability of the different protein-stabilized emulsions was in the order of casein > WPI > SPI as determined by monitoring both lipid hydroperoxide and headspace hexanal formation. The degree of positive charge on the protein-stabilized emulsion droplets was not the only factor involved in the inhibition of lipid oxidation because the charge of the emulsion droplets (WPI > casein > or = SPI) did not parallel oxidative stability. Other potential reasons for differences in oxidative stability of the protein-stabilized emulsions include differences in interfacial film thickness, protein chelating properties, and differences in free radical scavenging amino acids. This research shows that differences can be seen in the oxidative stability of protein-stabilized emulsions; however, further research is needed to determine the mechanisms for these differences.     

Lignin degradation by Agaricus bisporus accounts for a 30% increase in bioavailable holocellulose during cultivation on compost

The common mushroom Agaricus bisporus is a non-white rot saphrophytic fungus that can degrade lignin to free and utilize holocellulose embedded in fermented straw as present in compost. A new method is described to estimate the actual amount of bioavailable holocellulose in 3.8 kg compost cultures spawned with A. bisporus Horst U1 prior to and during a cultivation with two cycles of mushroom harvesting. The method shows that the initial amount of bioavailable holocellulose per culture, accounting for 130 +/- 22 g, is lower than the total holocellulose consumption by A. bisporus accounting for 182 +/- 15 g. This difference is explained by a 30% increase in bioavailable holocellulose. The increase is caused by the degradation of 95 +/- 3 g of holocellulose-shielding lignin. The results are discussed within the scope of the A. bisporus mushroom yield and lignin degradation by white rot fungi during growth on lignocellulose-containing materials.     

Production and characterization of O/W emulsions containing cationic droplets stabilized by lecithin-chitosan membranes

Oil-in-water emulsions containing cationic droplets stabilized by lecithin-chitosan membranes were produced using a two-stage process. A primary emulsion was prepared by homogenizing 5 wt % corn oil with 95 wt % aqueous solution (1 wt % lecithin, 100 mM acetic acid, pH 3.0) using a high-pressure valve homogenizer. This emulsion was diluted with aqueous chitosan solutions to form secondary emulsions with varying compositions: 1 wt % corn oil, 0.2 wt % lecithin, 100 mM acetic acid, and 0-0.04 wt % chitosan (pH 3.0). The particle size distribution, particle charge, and creaming stability of the primary and secondary emulsions were measured. The electrical charge on the droplets increased from -49 to +54 mV as the chitosan concentration was increased from 0 to 0.04 wt %, which indicated that chitosan adsorbed to the droplet surfaces. The mean particle diameter of the emulsions increased dramatically and the emulsions became unstable to creaming when the chitosan concentration exceeded 0.008 wt %, which was attributed to charge neutralization and bridging flocculation effects. Sonication, blending, or homogenization could be used to disrupt flocs formed in secondary emulsions containing droplets with high positive charges, leading to the production of emulsions with relatively small particle diameters (approximately 1 microm). These emulsions had good stability to droplet aggregation at low pH (< or =5) and ionic strengths (<500 mM). The interfacial engineering technology utilized in this study could lead to the creation of food emulsions with improved stability to environmental stresses.     

FUM13 encodes a short chain dehydrogenase/reductase required for C-3 carbonyl reduction during fumonisin biosynthesis in Gibberella moniliformis

Fumonisins are polyketide-derived mycotoxins produced by the filamentous fungus Gibberella moniliformis (anamorph Fusarium verticillioides). Wild-type strains of the fungus produce predominantly four B-series fumonisins, designated FB(1), FB(2), FB(3), and FB(4). Recently, a cluster of 15 putative fumonisin biosynthetic genes (FUM) was described in G. moniliformis. We have now conducted a functional analysis of FUM13, a gene in the cluster that is predicted by amino acid sequence similarity to encode a short chain dehydrogenase/reductase (SDR). Mass spectrometric analysis of metabolites from FUM13 deletion mutants revealed that they produce approximately 10% of wild-type levels of B-series fumonisins as well as two previously uncharacterized compounds. NMR analysis revealed that the new compounds are similar in structure to FB(3) and FB(4) but that they have a carbonyl function rather than a hydroxyl function at carbon atom 3 (C-3). These results indicate that the FUM13 protein catalyzes the reduction of the C-3 carbonyl to a hydroxyl group and are the first biochemical evidence directly linking a FUM gene to a specific reaction during fumonisin biosynthesis. The production of low levels of FB(1), FB(2), FB(3), and FB(4), which have a C-3 hydroxyl, by the FUM13 mutants suggests that G. moniliformis has an additional C-3 carbonyl reductase activity but that this enzyme functions less efficiently than the FUM13 protein.     

Polybrominated diphenyl ethers (PBDEs) in foodstuffs: human exposure through the diet

Polybrominated diphenyl ethers (PBDEs) are used as flame retardants in a variety of materials, including synthetic polymers and textiles. Although these chemicals have been detected in environmental samples and human tissues, there is little information about human exposure to PBDEs through the diet. In the present study, we determined the concentrations of PBDEs in a number of food samples acquired in Catalonia (Spain) during 2000. The dietary intake of PBDEs was estimated for the general population living in this Spanish region. The highest PBDE concentrations were found in oils and fats, fish and shellfish, meat and meat products, and eggs, while the lowest levels corresponded to fruits, vegetables, and tubers. The dietary intake of PBDEs for an adult male was 97.3 ng/day (assuming not detected (ND) = (1)/(2) limit of detection (LOD)) or 81.9 ng/day (assuming ND = 0) The greatest contribution to these values corresponded to fish and shellfish, with approximately one-third of the total intake. TetraBDEs and pentaBDEs were the homologues showing the highest percentages of contribution to the sum of total PBDEs. The comparison of the current dietary intake with the suggested lowest observed adverse effect level value of 1 mg/kg/day for the most sensitive endpoints for toxic effects of PBDEs results in a safety factor over 5 orders of magnitude in relation to PBDE exposure from food.