Vertical advection of extracellular DNA by water capillarity in soil columns

The fate of extracellular DNA in the environment concerns both the fate of transgenes from genetically modified organisms and the evolution of active bacteria capable of incorporating this DNA into their genomes. This study addressed the possibility that DNA, like other organic molecules, could move vertically in the capillary fringe of groundwater aquifers. The targeted gene fragment used here was the 35S-nptII sequence, which was below detection levels in controls. Initial microcosm studies detected the DNA target molecule by PCR during the entire experiment. The vertical advection of water and DNA were monitored for a period of 3 days in soil columns. DNA was added as a water solution at the bottom of the unsaturated soil column, and then DNA-free water was added at the bottom after 12 and 24 h. After the addition of the DNA solution, capillary water rose 4 cm within the soil column and the target DNA was detected up to that height. After 60 min, the entire soil column (10 cm) was wetted and the target sequence was detected up to a height of 7.5 cm. After the second wetting (12 h later), the target sequence was detected up to the top of the soil column (10 cm). However, after the third wetting (24 h later), the marker sequence was only found at heights from 0.5 to 4 cm. Results clearly show the vertical movement of DNA due the capillary rise and suggest the possibility of DNA degradation within the soil column.     

Ion Leaching Before and After Clear-Cutting in a Norway Spruce Stand - Effects of Long-Term Application of Ammonium Nitrate and Superphosphate

Soil solution chemistry was measured in a Haplic Podzol in a Norway spruce (Picea abies, L.Karst) site in central Sweden that had received frequent low-dose applications of nitrogen (N) asammonium nitrate (30-120 kg N/ha yr) and phosphorus (P) as superphosphate (20-40 kg P/haabout every third year) over a long period (22 yr). The aim was to investigate effects of added Non the ion leaching before and after clear-cutting. The N load corresponded well with actual loadsin central Europe. Prior to clear-cutting, fertilization increased concentrations of several ions inthe soil solution. Total losses of NO ₃ ^- , Al andH⁺ increased with the intensity of fertilization. However, since fertilization enhanced tree growthand nutrient uptake, differences between the total losses of several other ions tended to beequalized. After clear-cutting, the pH in the soil solution decreased, and the losses of Mn and Alincreased with fertilizer load, probably because of increased N mineralization and nitrification. Inthe N treatments the concentration of SO ₄ ^2- in the soil solution decreased, probably owing toincreased adsorption in the B horizon in response to the pH decrease. The effects ofanthropogenic N deposition are discussed in the light of our findings and possible causes oftemporal and spatial variation in soil solution chemistry and ion fluxes are suggested.     

Genetic diversity in sweetpotato [Ipomoea batatas (L.) Lam.] in relationship to geographic sources as assessed with RAPD markers

Available evidence shows that sweetpotato originated from either Central or South American lowlands with subsequent dispersal to North America, Europe, Africa, Asia and the pacific islands. A total of 71 polymorphic RAPD molecular markers were used to assess the genetic relationships amongst 74 sweetpotato varieties originating from a total of 23 sweetpotato producing countries within six geographical regions, namely, South America, Central America/Caribbean, United States of America (USA), East Africa, Asia and Oceania. An Analysis of Molecular Variance (AMOVA) indicated that 93.4% of the total variance was due to the differences between genotypes within regions. The difference between regions was significant (P < 0.001) but only contributed 6.6% to the variance. Genetic distance (PhiST) calculated with AMOVA and multidimensional scaling (MDS) revealed that the South American and the Central American/Caribbean genotypes formed two separate clusters. East African varieties, which have unique characteristics from other traditional varieties, were distinct from other traditional varieties from South America and Oceania. These results support the reported hypothesis of the origin and dispersal of the sweetpotato and indicate that the primary centre of diversity probably has two distinct genepools. It is proposed that the dispersal of the sweetpotato from its origin may have mainly involved varieties from Central America/Caribbean as opposed to varieties from South America. There is an indication that new genepools may be evolving in Africa and Asia due to hybridisation and adaptation to the local environments.     

Aluminium speciation: Variations caused by the choice of analytical method and by sample storage

We have compared a new FIA method (M1) for aluminium speciation in natural waters and a manual one based on one of Driscoll's proposals (M2). In synthetic solutions, aluminium's fluoro complexes were measured as ‘labile monomeric AP (Al _i ) by M2, but not as ‘quickly reacting Al’ (Al _qr ) by M1. Aluminium's complexes with Nordic Reference Humic Acid were measured neither as Al _i nor as Al _qr , and the same result was obtained for Al's citrato complexes. After excluding aluminium's fluoro complexes from Al _i , the results of the two methods agreed well for soil leaching samples and fairly well for natural water samples. Detection limits: 10μg L^?1 (ca. 0.4 μM) for both methods. Sample throughputs: M1∶66 injections h^?1; M2∶5 samples h^?1. Repeatabilities (RSD) on natural water samples: M1∶0.6–5.8% and M2 0.7–4.6%. Sample storage studies on soil solutions (FIA method) indicated that storage effects were ‘sample type specific’. A sample with a low level of Al _qr and a high DOC level (P2A) was more sensitive to storage than one with a high [Al] _qr and a low [DOC] (P2B^*). The decrease in [Al] _qr was statistically significant after 10 h (P2A) and 24 h (P2B^*), respectively. After 3 days' storage, [Al] _qr had decreased by some 40% in P2A, while the decrease in P2B^* was less than 10% after 8 days. The results of this part of the study also emphasize the importance of careful method standardization in sample storage studies.     

Nitrogen metabolism in flag leaf and grain of wheat in response to irrigation regimes

Although scarcity of irrigation water is one of the key limiting factors for wheat production in many regions of the world, by using partial irrigation at strategic times during the growing season, it might be possible to enhance productivity. We measured the changes in various parameters related to nitrogen (N) metabolism in flag leaf and grain of wheat (Triticum aestivum L.) plants (cv. Jinan 17 and Lumai 21), which were subjected to five irrigation regimes until physiological maturity. Severely deficient or excessive irrigation during grain filling decreased the photosynthetic rate (A), the concentrations of N, free amino acid, and soluble protein, as well as the activities of nitrate reductase (NR) and glutamine synthetase (GS) and increased malondialdehyde (MDA) accumulation and endopeptidase (EP) activity, though grain protein concentration might mainly depend on genotype. The activities of NR and GS were significantly positively correlated with A, but those of EP were significantly negatively correlated with A. The results indicate that while severe water stress aggravates the adverse effect on nitrogen metabolism, excessive soil moisture is also not useful during the grain‐filling stage, resulting in lower grain yield and quality. Our results suggest that applying an optimal irrigation regime in wheat fields still plays an important role in the improvement of grain yield and quality.     

Soil‐ and plant‐based nitrogen‐fertilizer recommendations in arable farming

Under‐ as well as overfertilization with nitrogen (N) will result in economic loss for the farmer due to reduced yields and quality of the products. Also from an ecological perspective, it is important that the grower makes the correct decision on how much and when to apply N for a certain crop to minimize impacts on the environment. To aggravate the situation, N is a substance that is present in many compartments in different forms (nitrate, ammonium, organic N, etc.) in the soil‐plant environment and takes part in various processes (e.g., mineralization, immobilization, leaching, denitrification, etc.). Today, many N‐recommendation systems are mainly based on yield expectation. However, yields are not stable from year to year for a given field. Also the processes that determine the N supply from other sources than fertilizer are not predictable at the start of the growing season. Different methodological approaches are reviewed that have been introduced to improve N‐fertilizer recommendations for arable crops. Many soil‐based methods have been developed to measure soil mineral N (SMN) that is available for plants at a given sampling date. Soil sampling at the start of the growing period and analyzing for the amount of NO ‐N (and NH ‐N) is a widespread approach in Europe and North America. Based on data from field calibrations, the SMN pool is filled up with fertilizer N to a recommended amount. Depending on pre‐crop, use of organic manure, or soil characteristics, the recommendation might be modified (±10–50 kg N ha^–1). Another set of soil methods has been established to estimate the amount of N that is mineralized from soil organic matter, plant residues, and/or organic manure. From the huge range of methods proposed so far, simple mild extraction procedures have gained most interest, but introduction into practical recommendation schemes has been rather limited. Plant‐analytical procedures cover the whole range from quantitative laboratory analysis to semiquantitative “quick” tests carried out in the field. The main idea is that the plant itself is the best indicator for the N supply from any source within the growth period. In‐field methods like the nitrate plant sap/petiole test and chlorophyll measurements with hand‐held devices or via remote sensing are regarded as most promising, because with these methods an adequate adjustment of the N‐fertilizer application strategy within the season is feasible. Prerequisite is a fertilization strategy that is based on several N applications and not on a one‐go approach.     

In vitro inhibition of the activation of Pro-matrix Metalloproteinase 1 (Pro-MMP-1) and Pro-matrix metalloproteinase 9 (Pro-MMP-9) by rice and soybean Bowman-Birk inhibitors

The in vitro inhibitory activity of the rice Bowman-Birk inhibitor (rBBI) or soybean Bowman-Birk inhibitor (sBBI) against trypsin-catalyzed activation of pro-matrix metalloproteinase 1 or 9 (pro-MMP-1 or pro-MMP-9), respectively, was investigated using electrophoresis with silver staining, heparin-enhanced zymography, biotinylated gelatin, Biotrak assay, and fluorescence quenched substrate hydrolysis. rBBI at concentrations of 0.08-0.352 mg/mL dose-dependently inhibited the in vitro activation of 45 microg/mL pro-MMP-1 by trypsin. Heparin-enhanced zymography analysis of pro-MMP-1, trypsin-activated MMP-1, and a mixture of pro-MMP-1-trypsin-rBBI showed clear zones associated with trypsin-activated MMP-1 and the absence of clear zones in lanes containing pro-MMP-1 or a mixture of pro-MMP-1, trypsin, and rBBI. The results of the Biotrak assay also indicated that rBBI dose-dependently suppressed the activation of pro-MMP-1 by trypsin. sBBI dose-dependently inhibited the activation of 100 microg/mL of pro-MMP-9 by trypsin. Biotinylated gelatin assays demonstrated that pro-MMP-9 or pro-MMP-9 in the presence of trypsin and BBI did not hydrolyze gelatin, whereas p-aminophenylmercury acetate (APMA)-activated MMP-9 and trypsin-activated MMP-9 caused significant hydrolysis of gelatin. Quenched fluorescence substrate hydrolysis for total MMP activity showed that pro-MMP-1 or pro-MMP-9 did not hydrolyze the substrate Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2; active MMP-1 or MMP-9 hydrolyzed the substrate, but lower substrate hydrolysis was obtained when pro-MMP-1 or pro-MMP-9 was incubated with trypsin in the presence of increasing concentrations of rBBI. The results are discussed in light of the role of MMP-1 and MMP-9 in the process of angiogenesis and the potential of rBBI or sBBI as a functional food ingredient.     

Antiradical activity of water soluble components in common diet vegetables

The antiradical activity of water-soluble components contained in mushrooms (Psalliota campestris), onions (Allium cepa), white cabbage (Brassica oleracea var. alba), and yellow bell peppers (Capsicum annuum) against hydroxyl radicals was tested in a chemical and biological system. The vegetable juices were obtained by centrifugation of a vegetable homogenate processed at 2 degrees C or heated at 102 degrees C. The chemical system consisted of a buffered reaction mixture composed of Fe(III)-EDTA, 2-deoxy-D-ribose, ascorbic acid, and H(2)O(2) generating the hydroxyl radical. The antiradical activity was expressed as an inhibition of deoxyribose degradation. The biological system consisted of IMR32 neuroblastoma cells exposed to H(2)O(2) in the presence or absence of the vegetable juices. Cells were pretreated for either 24 h or 1 h with the vegetable juices, and reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was used as a cell viability assay. All vegetable juices inhibited the degradation of deoxyribose and increased the viability of H(2)O(2) treated cells. Raw mushroom juice proved to be the most active in both cases. Boiling significantly affected the activity of mushroom juice, but did not change significantly the effect on onions and yellow bell peppers, and partially increased the activity of white cabbage juice. Mushroom antiradical activity was also confirmed by a cytofluorimetric analysis.     

Chemical characterization and biological effects of Sicilian Opuntia ficus indica (L.) mill. Fruit juice: antioxidant and antiulcerogenic activity

The juice of whole fruits of Sicilian cultivars of prickly pear (Opuntia ficus indica (L.) Mill.) was investigated, and the contents of ascorbic acid, total polyphenols, and flavonoids were determined. In the juice, ferulic acid was the chief derivative of hydroxycinnamic acid and the mean concentration of total phenolic compounds was 746 microg/mL. The flavonoid fraction, analyzed by high-performance liquid chromatography-diode array detection, consisted of rutin and isorhamnetin derivatives. The juice showed antioxidant activity in the DPPH(*) test, probably due to the phenolic compounds that are effective radical scavengers. The preventive administration of the juice inhibited the ulcerogenic activity of ethanol in rat. Light microscopy observations showed an increase in mucus production and the restoration of the normal mucosal architecture. The juice is nutritionally interesting, and its dietary intake could provide protection against oxidative damage.