Effect of Brazilian spinach (Alternanthera sissoo) pellet supplementation and dietary ratios on rumen characteristics, microorganisms, methane production, milk yield, and milk composition in dairy cows.

The aim of the previous research was to evaluate the effects of Brazilian spinach pellet (BSP) supplementation and dietary ratios on rumen characteristics, methane estimation, and milk production in dairy cows. Four crossbred Thai dairy cattle, with Holstein Friesian (HF) cows with a body weight of 442 ± 50 kg were assessed in a 2 × 2 factorial in a 4 × 4 Latin square design to obtain diets; factor A was the roughage (R) to concentrate (C) ratio at 40:60 and 30:70, and factor B was level of BSP supplantation at 2% and 6% of dry matter (basis) intake (DMI). R:C ratio and supplementation of BSP had no interaction effect on DMI and nutrient digestibility. On DM, organic matter (OM), crude protein (CP), and acid detergent fiber (ADF) intake, the R:C ratio increased (p < 0.05). The digestibility of OM improved (p < 0.05) when cows were fed a R:C ratio of 30:70. On pH, ammonia-nitrogen, protozoal population, and blood urea-nitrogen, there were no interactions between the R:C ratio and BSP supplementation. Increasing the BSP supplementation to 6% (p < 0.01) decreased the protozoal population. The R:C ratio of 30:70 increased total volatile fatty acid (VFA) and propionate (C3) concentrations while decreasing the acetate (C2) to C3 ratio and methane (CH4 ) estimation (p < 0.01). The average concentration of total VFA has increased by 114.46 mmol/L for 6% of BSP supplementation. Increased BSP supplementation increased the C3 concentration while decreasing the C2:C3 ratio and CH4 emissions (p < 0.05). The R:C ratio and BSP supplementation had no interaction effect on milk yield, 3.5% fat-corrected milk (FCM), or milk composition. The R:C ratio of 30:70 increased milk yield (p < 0.05) to the highest level of 12.18 kg/day. In conclusion, the diet containing a R:C ratio of 30:70 increased feed intake, milk yield, BUN, total VFA, and C3 concentration, and decreased the C2:C3 ratio and CH4 emission. BSP supplementation at 6% could increase TVFA and C3 concentrations while decreasing the protozoal population and CH4 estimation.

Selective accumulation of pharmaceutical residues from 6 different soils by plants: a comparative study on onion, radish, and spinach.

The accumulation of six pharmaceuticals of different therapeutic uses has been thoroughly investigated and compared between onion, spinach, and radish plants grown in six soil types. While neutral molecules (e.g., carbamazepine (CAR) and some of its metabolites) were efficiently accumulated and easily translocated to the plant leaves (onion > radish > spinach), the same for ionic (both anionic and cationic) molecules seems to be minor to moderate. The maximum accumulation of CAR crosses 38,000 (onion), 42,000 (radish), and 7000 (spinach) ng g-1 (dry weight) respectively, in which the most majority of them happened within the plant leaves. Among the metabolites, the accumulation of carbamazepine 10,11-epoxide (EPC - a primary CAR metabolite) was approximately 19,000 (onion), 7000 (radish), and 6000 (spinach) ng g-1 (dry weight) respectively. This trend was considerably similar even when all these pharmaceuticals applied together. The accumulation of most other molecules (e.g., citalopram, clindamycin, clindamycin sulfoxide, fexofenadine, irbesartan, and sulfamethoxazole) was restricted to plant roots, except for certain cases (e.g., clindamycin and clindamycin sulfoxide in onion leaves). Our results clearly demonstrated the potential role of this accumulation process on the entrance of pharmaceuticals/metabolites into the food chain, which eventually becomes a threat to associated living biota.

Phytoextraction of nickel, lead, and chromium from contaminated soil using sunflower, marigold, and spinach: comparison of efficiency and fractionation study.

Heavy metals in soil pose a serious threat through their toxic effect on the human food chain. Phytoremediation is a clean and green potentially cost-effective technology in remediating the heavy metal-contaminated soil. However, the efficiency of phytoextraction is very often limited by low phytoavailability of heavy metals in soil, slow growth, and small biomass production of hyper-accumulator plants. To solve these issues, accumulator plant(s) with high biomass production and amendment(s) which can solubilize metals in soil is required for better phytoextraction. A pot experiment was conducted to assess the efficiency of phytoextraction of sunflower, marigold, and spinach as affected by the incorporation of Sesbania (solubilizer) and addition of gypsum (solubilizer) in nickel (Ni)-, lead (Pb)-, and chromium (Cr)-contaminated soil. A fractionation study was conducted to study the bioavailability of the heavy metals in contaminated soil after growing the accumulator plants and as affected by using soil amendments (Sesbania and gypsum). Results showed that marigold was the most efficient among the three accumulator plants in phytoextraction of the heavy metals in the contaminated soil. Both sunflower and marigold were able to reduce the bioavailability of the heavy metals in the post-harvest soil, which was reflected in their (heavy metals) lower concentration in subsequently grown paddy crop (straw). The fractionation study revealed that carbonate and organically bound fractions of the heavy metals control the bioavailability of the heavy metals in the experimental soil. Both Sesbania and gypsum were not effective in solubilizing the heavy metals in the experimental soil. Therefore, the possibility of using Sesbania and gypsum for solubilizing heavy metals in contaminated soil is ruled out.

Silicon fertilization counteracts salinity-induced damages associated with changes in physio-biochemical modulations in spinach.

Plant growth and productivity are limited by the severe impact of salt stress on the fundamental physiological processes. Silicon (Si) supplementation is one of the promising techniques to improve the resilience of plants under salt stress. This study deals with the response of exogenous Si applications (0, 2, 4, and 6 mM) on growth, gaseous exchange, ion homeostasis and antioxidant enzyme activities in spinach grown under saline conditions (150 mM NaCl). Salinity stress markedly reduced the growth, physiological, biochemical, water availability, photosynthesis, enzymatic antioxidants, and ionic status in spinach leaves. Salt stress significantly enhanced leaf Na+ contents in spinach plants. Supplementary foliar application of Si (4 mM) alleviated salt toxicity, by modulating the physiological and photosynthetic attributes and decreasing electrolyte leakage, and activities of SOD, POD and CAT. Moreover, Si-induced mitigation of salt stress was due to the depreciation in Na+/K+ ratio, Na+ ion uptake at the surface of spinach roots, and translocation in plant tissues, thereby reducing the Na+ ion accumulation. Foliar applied Si (4 mM) ameliorates ionic toxicity by decreasing Na+ uptake. Overall, the results illustrate that foliar applied Si induced resistance against salinity stress in spinach by regulating the physiology, antioxidant metabolism, and ionic homeostasis. We advocate that exogenous Si supplementation is a practical approach that will allow spinach plants to recover from salt toxicity.

Natural particles can armor emulsions against lipid oxidation and coalescence.

Traditional functional ingredients, such as conventional emulsifiers (surfactants, animal-derived proteins), and synthetic antioxidants may become obsolete in the development of clean-label, plant-based, sustainable food emulsions. Previously, we showed that tailor-made antioxidant-loaded particles can yield both physically and oxidatively stable emulsions, and we expected that natural particles with related properties could also show these beneficial effects. Here, we investigated Pickering emulsions prepared with natural plant particulate materials. Particles that showed weak aggregation in acidic aqueous media, indicating a relatively hydrophobic surface, were able to physically stabilize oil-in-water emulsions, through either Pickering stabilization (powders of matcha tea, spinach leaves, and spirulina cake), or an increase in viscosity (pineapple fibers). Matcha tea and spinach leaf particle-stabilized emulsions were highly stable to lipid oxidation, as compared to emulsions stabilized by conventional emulsifiers. Taking this dual particle functionality as a starting point for emulsion design is, in our view, essential to achieve clean-label food emulsions.

Plant terpenoid metabolism co-opts a component of the cell wall biosynthesis machinery.

Glycosylation is one of the most prevalent molecular modifications in nature. Single or multiple sugars can decorate a wide range of acceptors from proteins to lipids, cell wall glycans and small molecules, dramatically affecting their activity. Here, we discovered that by 'hijacking' an enzyme of the cellulose synthesis machinery involved in cell wall assembly, plants evolved cellulose synthase-like enzymes (Csls) and acquired the capacity to glucuronidate specialized metabolites, that is, triterpenoid saponins. Apparently, endoplasmic reticulum-membrane localization of Csls and of other pathway proteins was part of evolving a new glycosyltransferase function, as plant metabolite glycosyltransferases typically act in the cytosol. Discovery of glucuronic acid transferases across several plant orders uncovered the long-pursued enzymatic reaction in the production of a low-calorie sweetener from licorice roots. Our work opens the way for engineering potent saponins through microbial fermentation and plant-based systems.

In vitro digestion of galactolipids from chloroplast-rich fraction (CRF) of postharvest, pea vine field residue (haulm) and spinach leaves.

The removal of intact chloroplasts from their cell wall confinement offers a novel way to obtain lipophilic nutrients from green biomass, especially carotenoids and galactolipids. These latter are the main membrane lipids in plants and they represent a major source of the essential α-linolenic acid (18:3; ALA). Nevertheless, knowledge on their digestion is still limited. We have developed a physical method of recovering a chloroplast-rich fraction (CRF) from green biomass and tested its digestibility in vitro under simulated gastrointestinal conditions. Using a two-step static model, CRF from both spinach leaves and postharvest, pea vine field residue (haulm) were first exposed to enzymes from rabbit gastric extracts and then either to pancreatic enzymes from human pancreatic juice (HPJ) or to porcine pancreatic extracts (PPE). The lipolysis of monogalactosyldiacylglycerol (MGDG) and digalactosyl diacylglycerol (DGDG) was monitored by thin layer chromatography and gas chromatography of fatty acid methyl esters. For both CRF preparations, MGDG and DGDG were converted to monogalactosylmonoacylglycerol (MGMG) and digalactosylmonoacylglycerol (DGMG), respectively, during the intestinal phase and ALA was the main fatty acid released. Galactolipids were more effectively hydrolysed by HPJ than by PPE, and PPE showed a higher activity on MGDG than on DGDG. These findings may be explained by the higher levels of galactolipase activity in HPJ compared to PPE, which mainly results from pancreatic lipase-related protein 2. Thus, we showed that CRF galactolipids are well digested by pancreatic enzymes and represent an interesting vehicle for ALA supplementation in human diet.

Improvement of carotenoid bioaccessibility from spinach by co-ingesting with excipient nanoemulsions: impact of the oil phase composition.

Many of the carotenoids found naturally in fruits and vegetables are beneficial to human health, but they often have low oral bioavailability because of their high hydrophobicity. In this study, the effects of varying the composition of the oil phase of excipient nanoemulsions on carotenoid bioaccessibility from spinach were investigated using a simulated gastrointestinal tract. Nanoemulsions containing different ratios of medium chain triglycerides (MCT) and long chain triglycerides (LCT) were prepared: (i) mixing MCT and LCT oils before homogenization and (ii) mixing MCT droplets with LCT droplets after homogenization. The release of carotenoids from spinach and their solubilization within the mixed micelles formed after lipid digestion depended strongly on the oil phase composition. As expected, carotenoid bioaccessibility was always higher in the presence of excipient nanoemulsions than in their absence. The total free fatty acids released in the small intestine increased as the MCT/LCT ratio increased, which can be attributed to the faster release of shorter chain fatty acids from the oil droplet surfaces during lipid digestion. As the MCT ratio increased, lutein bioaccessibility increased but ß-carotene bioaccessibility decreased. This difference was attributed to the ability of the formed mixed micelles to accommodate the two different kinds of carotenoids in their hydrophobic domains. Interestingly, carotenoid bioaccessibility was significantly lower (P < 0.05) when the oil droplets were mixed after homogenization than when the oils were mixed before homogenization. These results have important implications for the design of excipient foods to improve the bioavailability of hydrophobic nutraceuticals in fruits and vegetables.

Single-molecule study of redox control involved in establishing the spinach plastocyanin-cytochrome b6f electron transfer complex.

Small diffusible redox proteins play a ubiquitous role in bioenergetic systems, facilitating electron transfer (ET) between membrane bound complexes. Sustaining high ET turnover rates requires that the association between extrinsic and membrane-bound partners is highly specific, yet also sufficiently weak to promote rapid post-ET separation. In oxygenic photosynthesis the small soluble electron carrier protein plastocyanin (Pc) shuttles electrons between the membrane integral cytochrome b6f (cytb6f) and photosystem I (PSI) complexes. Here we use peak-force quantitative nanomechanical mapping (PF-QNM) atomic force microscopy (AFM) to quantify the dynamic forces involved in transient interactions between cognate ET partners. An AFM probe functionalised with Pc molecules is brought into contact with cytb6f complexes, immobilised on a planar silicon surface. PF-QNM interrogates the unbinding force of the cytb6f-Pc interactions at the single molecule level with picoNewton force resolution and on a time scale comparable to the ET time in vivo (ca. 120 µs). Using this approach, we show that although the unbinding force remains unchanged the interaction frequency increases over five-fold when Pc and cytb6f are in opposite redox states, so complementary charges on the cytb6f and Pc cofactors likely contribute to the electrostatic forces that initiate formation of the ET complex. These results suggest that formation of the docking interface is under redox state control, which lowers the probability of unproductive encounters between Pc and cytb6f molecules in the same redox state, ensuring the efficiency and directionality of this central reaction in the 'Z-scheme' of photosynthetic ET.

Transient Expression in Red Beet of a Biopharmaceutical Candidate Vaccine for Type-1 Diabetes.

Plant molecular farming is the use of plants to produce molecules of interest. In this perspective, plants may be used both as bioreactors for the production and subsequent purification of the final product and for the direct oral delivery of heterologous proteins when using edible plant species. In this work, we present the development of a candidate oral vaccine against Type 1 Diabetes (T1D) in edible plant systems using deconstructed plant virus-based recombinant DNA technology, delivered with vacuum infiltration. Our results show that a red beet is a suitable host for the transient expression of a human derived autoantigen associated to T1D, considered to be a promising candidate as a T1D vaccine. Leaves producing the autoantigen were thoroughly characterized for their resistance to gastric digestion, for the presence of residual bacterial charge and for their secondary metabolic profile, giving an overview of the process production for the potential use of plants for direct oral delivery of a heterologous protein. Our analysis showed almost complete degradation of the freeze-dried candidate oral vaccine following a simulated gastric digestion, suggesting that an encapsulation strategy in the manufacture of the plant-derived GAD vaccine is required.

Cadmium uptake by onions, lettuce and spinach in New Zealand: Implications for management to meet regulatory limits.

Paired soil and plant samples collected from the main commercial growing areas for onions (Allium cepa), lettuce (Lactuca sativa) and spinach (Spinacia olearacea) in New Zealand were used to assess the influence of plant and soil factors on cadmium (Cd) uptake in these crops. Differences in Cd concentration between eight lettuce sub-types were not consistent across sites, nor were differences in Cd concentrations in three crisphead cultivars assessed at two sites. Similarly, differences in Cd concentrations between four onion cultivars were inconsistent across sites. Mean lettuce Cd concentrations in eight lettuce varieties (range 0.005-0.034 mg∙kg-1 (fresh weight, FW) were markedly lower than those in baby leaf and bunching spinach, (range 0.005-0.19 mg∙kg-1 FW). Significant regional variation was observed in Cd concentrations in one onion cultivar (mean range 0.007-0.05 mg∙kg-1 FW). Soil Cd concentration, pH and region were statistically significant predictors of onion Cd concentration, explaining low (38% for soil Cd and pH) to moderate (50% for all three parameters) percentage of the variation. Soil Cd concentration and exchangeable magnesium or total carbon were statistically significant predictors of Cd concentration in baby leaf and bunching spinach, respectively, explaining a moderate percentage (49% and 42%) of the variation in Cd concentration. Increasing pH and soil carbon may assist in minimising Cd uptake in onion and bunching spinach, respectively. The low to moderate proportion of explained variation is partly attributable to the narrow range in some measured soil properties and indicates factors other than those assessed are influencing plant uptake. This highlights a challenge in using these relationships to develop risk-based soil guideline values to support compliance with food standards. Similarly, the inconsistency in Cd concentrations in different cultivars across sites highlights the need for multi-site assessments to confirm the low Cd accumulation status of different cultivars.

Enhancement of phytochemical bioaccessibility from plant-based foods using excipient emulsions: impact of lipid type on carotenoid solubilization from spinach.

Effects of lipid type in excipient emulsions on the bioaccessibility of carotenoids (lutein and ß-carotene) in spinach were studied using a simulated gastrointestinal tract (GIT). Results showed that the lipid type only had a minor impact on the physical and structural characteristics of the spinach/emulsion mixtures as they passed through simulated mouth, stomach, and small intestine phases. However, a significant effect was observed on lipid digestion, mixed micelle formation, and carotenoid bioaccessibility. Excipient emulsions containing mainly medium chain triacylglycerols (MCTs) (MCT and coconut oils) had faster initial lipid digestion rates, higher overall digestibility, smaller mixed micelle sizes, and higher lutein bioaccessibilities than those containing mainly long chain triacylglycerols (LCTs) (corn, olive, and fish oils). Excipient emulsions rich in long chain monounsaturated fatty acids (MUFAs) (corn and olive oils) formed larger mixed micelles and gave higher ß-carotene bioaccessibilities than those rich in either medium chain saturated fatty acids (SFAs) (MCT and coconut oils) or long chain polyunsaturated fatty acids (PUFAs) (fish oil). These differences in bioaccessibility were attributed to differences in micelle size and solubilization capacity, as well as carotenoid dimensions. Finally, emulsions containing a mixed oil phase (MCT oil : corn oil = 1 : 1, w/w) appreciably increased both lutein (from 21% to 42%) and ß-carotene (from 6.8% to 25%) bioaccessibility from spinach compared to a control (no oil). These results suggest that mixed LCT-MCT oil phases may be useful for the design of excipient emulsions for improving the bioaccessibility of various hydrophobic nutraceuticals.

Evaluation of the antimicrobial activities of ultrasonicated spinach leaf extracts using RAPD markers and electron microscopy.

Spinach (Spinacia oleracea L.) leaves represent an important dietary source of nutrients, antioxidants and antimicrobials. As such, spinach leaves play an important role in health and have been used in the treatment of human diseases since ancient times. Here, the aims were to optimize the extraction methods for recovering antimicrobial substances of spinach leaves, determine the minimum inhibitory concentrations (MICs) of the antimicrobial substances against Escherichia coli and Staphylococcus aureus and, finally, evaluate the effects of spinach leaves' antimicrobials on bacterial DNA using central composite face-centered methods. The effect of the extracts on both Gram-positive and Gram-negative bacterial models was examined by scanning electron microscopy (SEM) and random amplification of polymorphic (bacterial) DNA (RAPD). The optimal extraction conditions were at 45 °C, ultrasound power of 44% and an extraction time of 23 min. The spinach extracts exhibited antimicrobial activities against both bacteria with MICs in the 60-100 mg/ml range. Interestingly, SEM showed that the treated bacterial cells appear damaged with a reduction in cell number. RAPD analysis of genomic DNA showed that the number and sizes of amplicons were decreased by treatments. Based on these results, it was inferred that spinach leaf extracts exert bactericidal activities by both inducing mutations in DNA and causing cell wall disruptions.

Biochar enhances the cadmium tolerance in spinach (Spinacia oleracea) through modification of Cd uptake and physiological and biochemical attributes.

Cadmium (Cd) has no known role in plant biology and is toxic to plants and animals. The Cd mainly accumulated in agricultural soils through anthropogenic activities, such as sewage water irrigation and phosphorus fertilization. Biochar (BC) has been proposed as an amendment to reduce metal toxicity in plants. The objective of this study was to evaluate the role of BC (cotton stick at a rate of 0, 3, and 5 %) on Cd uptake and the photosynthetic, physiological, and biochemical responses of spinach (Spinacia oleracea) grown in Cd-spiked soil (0, 25, 50, 75, and 100 mg Cd kg-1 soil). The results showed that Cd toxicity decreased growth, photosynthetic pigments, gas exchange characteristics, and amino acid and protein contents in 52-day-old spinach seedlings. The Cd treatments increased the concentrations of Cd, sugar, ascorbic acid, and malondialdehyde (MDA) in plants. The application of BC ameliorated the harmful effects of Cd in spinach plants. Under Cd stress, BC application increased the growth, photosynthesis, and protein contents and decreased Cd concentrations and MDA contents in plants. The maximum BC-mediated increase in dry biomass was about 25 % with 5 % BC application in control plants. It is concluded that BC could ameliorate Cd toxic effects in spinach through changing the physiological and biochemical attributes under Cd stress.

Metabolomics for the Authentication of Natural Extracts Used in Flavors and Fragrances: the Case Study of Violet Leaf Absolutes from Viola odorata.

Natural extracts used in fine fragrances (alcoholic perfumes) are rare and precious. As such, they represent an interesting target for fraudulent practices called adulterations. Absolutes, important materials used in the creation of perfumes, are obtained by organic solvent extraction of raw plant materials. Because the nonvolatile part of these natural extracts is not normalized and scarcely reported, highlighting potential adulterations present in this fraction appears highly challenging. For the first time, we investigated the use of nontargeted UHPLC-ToFMS metabolomics for this purpose, considering Viola odorata l., a plant largely used in the perfume industry, as a model. Significant differences in the metabolic fingerprints of the violet leaf absolutes were evidenced according to geographical locations, and/or adulterations. Additionally, markers of the geographical origin were detected through their molecular weight/most probable molecular formula and retention time, while adulterations were statistically validated. In this study, we thus clearly demonstrated the efficiency of UHPLC-ToFMS-based metabolomics in accelerating both the identification of the origin of raw materials as well as the search for potential adulterations in absolutes, natural products of high added value.

Moderate water stress prevents the postharvest decline of ascorbic acid in spinach (Spinacia oleracea L.) but not in spinach beet (Beta vulgaris L.).

BACKGROUND: Babyleaf salads such as spinach (Spinacia oleracea L.) and spinach beet (Beta vulgaris L. subsp. cicla var. cicla) are an important dietary source of antioxidants such as ascorbic acid (vitamin C). Such compounds may be important in disease prevention in consumers but the level of these compounds in leaves frequently declines after harvest. As such, methods to maintain antioxidant levels in fresh produce are being sought. RESULTS: Irrigation deficits were used to apply water stress to S. oleracea and B. vulgaris plants. This treatment prevented postharvest decline of leaf ascorbic acid content in S. oleracea but not in B. vulgaris. Ascorbic acid levels in leaves at harvest were unaffected by the treatment in both species compared to well-watered controls. CONCLUSION: We have shown that restricted irrigation provides a viable means to maintain leaf vitamin content after harvest in S. oleracea, an important finding for producers, retailers and consumers alike. © 2015 Society of Chemical Industry.

Effects of phosphorus on chemical forms of Cd in plants of four spinach (Spinacia oleracea L.) cultivars differing in Cd accumulation.

In order to clarify how cadmium (Cd) chemical forms in planta relate to the genotype difference in Cd accumulation of spinach (Spinacia oleracea L.), two low-Cd and two high-Cd cultivars were compared under a hydroponic experiment with two concentrations of Cd (8.98 or 44.71 µmol Cd L(-1)). The concentrations of phosphorus in the hydroponic system were also adjusted to two levels (0.5 and 1.0 mmol L(-1)) to investigate the influence of phosphorus on the forms and accumulation of Cd in the tested cultivars. Average Cd concentrations in shoots were 8.50-10.06 mg kg(-1) for high-Cd cultivars and 6.11-6.64 mg kg(-1) for low-Cd cultivars a under lower Cd treatment and were as high as 24.41-31.35 mg kg(-1) and 19.65-25.76 mg kg(-1), respectively, under a higher treatment. Phosphorus significantly decreased Cd accumulation in the tested cultivars, and the effect had superiority over the cultivar alternation under higher Cd stress. Cadmium in the NaCl-extractable fraction of the plant tissues showed the greatest relationship to genotype difference of Cd accumulation. The difference in the capacity to binding Cd into F HAc, F HCl, or F Residue was another important mechanism involving in the genotype difference in Cd accumulation of spinach. Among them, average proportion of Cd in F HAc in low-Cd cultivars was higher than that in high-Cd cultivars in association with the effect of phosphorus.

Photodamage to the oxygen evolving complex of photosystem II by visible light.

Light damages photosynthetic machinery, primarily photosystem II (PSII), and it results in photoinhibition. A new photodamage model, the two-step photodamage model, suggests that photodamage to PSII initially occurs at the oxygen evolving complex (OEC) by light energy absorbed by manganese and that the PSII reaction center is subsequently damaged by light energy absorbed by photosynthetic pigments due to the limitation of electrons to the PSII reaction center. However, it is still uncertain whether this model is applicable to photodamage to PSII under visible light as manganese absorbs visible light only weakly. In the present study, we identified the initial site of photodamage to PSII upon illumination of visible light using PSII membrane fragments isolated from spinach leaves. When PSII samples were exposed to visible light in the presence of an exogenous electron acceptor, both PSII total activity and the PSII reaction centre activity declined due to photodamage. The supplemental addition of an electron donor to the PSII reaction centre alleviated the decline of the reaction centre activity but not the PSII total activity upon the light exposure. Our results demonstrate that visible light damages OEC prior to photodamage to the PSII reaction center, consistent with two-step photodamage model.

Bioremediation of petroleum hydrocarbon contaminated soil by Rhodobacter sphaeroides biofertilizer and plants.

Bio-augmentation is a promising technique for remediation of polluted soils. This study aimed to evaluate the bio-augmentation effect of Rhodobacter sphaeroides biofertilizer (RBF) on the bioremediation of total petroleum hydrocarbons (TPH) contaminated soil. A greenhouse pot experiment was conducted over a period of 120 days, three methods for enhancing bio-augmentation were tested on TPH contaminated soils, including single addition RBF, planting, and combining of RBF and three crop species, such as wheat (W), cabbage (C) and spinach (S), respectively. The results demonstrated that the best removal of TPH from contaminated soil in the RBF bio-augmentation rhizosphere soils was found to be 46.2%, 65.4%, 67.5% for W+RBF, C+RBF, S+RBF rhizosphere soils respectively. RBF supply impacted on the microbial community diversity (phospholipid fatty acids, PLFA) and the activity of soil enzymes, such as dehydrogenase (DH), alkaline phosphatase (AP) and urease (UR). There were significant difference among the soil only containing crude oil (CK), W, C and S rhizosphere soils and RBF bio-augmentation soils. Moreover, the changes were significantly distinct depended on crops species. It was concluded that the RBF is a valuable material for improving effect of remediation of TPH polluted soils.

Broad and efficient control of major foodborne pathogenic strains of Escherichia coli by mixtures of plant-produced colicins.

Enterohemorrhagic Escherichia coli (EHEC) is one of the leading causes of bacterial enteric infections worldwide, causing ∼100,000 illnesses, 3,000 hospitalizations, and 90 deaths annually in the United States alone. These illnesses have been linked to consumption of contaminated animal products and vegetables. Currently, other than thermal inactivation, there are no effective methods to eliminate pathogenic bacteria in food. Colicins are nonantibiotic antimicrobial proteins, produced by E. coli strains that kill or inhibit the growth of other E. coli strains. Several colicins are highly effective against key EHEC strains. Here we demonstrate very high levels of colicin expression (up to 3 g/kg of fresh biomass) in tobacco and edible plants (spinach and leafy beets) at costs that will allow commercialization. Among the colicins examined, plant-expressed colicin M had the broadest antimicrobial activity against EHEC and complemented the potency of other colicins. A mixture of colicin M and colicin E7 showed very high activity against all major EHEC strains, as defined by the US Department of Agriculture/Food and Drug Administration. Treatments with low (less than 10 mg colicins per L) concentrations reduced the pathogenic bacterial load in broth culture by 2 to over 6 logs depending on the strain. In experiments using meats spiked with E. coli O157:H7, colicins efficiently reduced the population of the pathogen by at least 2 logs. Plant-produced colicins could be effectively used for the broad control of pathogenic E. coli in both plant- and animal-based food products and, in the United States, colicins could be approved using the generally recognized as safe (GRAS) regulatory approval pathway.