Insecticidal activity of Arum maculatum tuber lectin and its binding to the glycosylated insect gut receptors

Mannose binding approximately 50 kDa homotetrameric lectin, purified from edible Arum maculatum tuber, was analyzed through SDS-PAGE and studied for its agglutination property using rabbit erythrocytes. Cross reactivity of the purified lectin was verified through western blot using Colocasia esculantum(Family, Araceae) tuber lectin antibody. The insecticidal activity of Arum maculatum tuber lectin (ATL) was tested against two economically important sucking pests, Lipaphis erysimi and Aphis craccivora, in an artificial diet. The LC(50) values for L. erysimi and A. craccivora were determined to be 21 microg/mL and 16 microg/mL, respectively. Addition of alpha-d-mannose in ATL-supplemented diet reduced the aphid mortality. Two major receptor proteins of ATL (approximately 40 kDa and approximately 35 kDa) were detected from the brush border membrane vesicle (BBMV) protein of L. erysimi and A. craccivora guts, respectively, using ligand-binding assay. Alpha-d-Mannose was found to be a deterrent to such binding of ATL to the BBMV receptors.     

Plant regeneration functional groups modulate the response to fire of soil enzyme activities in a Mediterranean shrubland

Soil enzymes are critical to soil nutrient cycling function but knowledge on the factors that control their response to major disturbances such as wildfires remains very limited. We evaluated the effect of fire-related plant functional traits (resprouting and seeding) on the resistance and resilience to fire of two soil enzyme activities involved in phosphorus and carbon cycling (acid phosphatase and β-glucosidase) in a Mediterranean shrublands in SE Spain. Using experimental fires, we compared four types of shrubland microsites: SS (vegetation patches dominated by seeder species), RR (patches dominated by resprouter species), SR (patches co-dominated by seeder and resprouter species), and IP (shrub interpatches). We assessed pre- and post-fire activities of the target soil enzymes, available P, soil organic C, and plant cover dynamics over three years after the fire. Post-fire regeneration functional groups (resprouter, seeder) modulated both pre- and post-fire activity of acid phosphatase and β-glucosidase, with higher activity in RR and SR patches than in SS patches and IP. However, we found no major differences in enzyme resistance and resilience between microsite types, except for a trend towards less resilience in SS patches. Fire similarly reduced the activity of both enzymes. However, acid phosphatase and β-glucosidase showed contrasting post-fire dynamics. While β-glucosidase proved to be rather resilient to fire, fully recovering three years after fire, acid phosphatase showed no signs of recovery in that period. Overall, the results indicate a positive influence of resprouter species on soil enzyme activity that is very resistant to fire. Long-lasting decrease in acid phosphatase activity probably resulted from the combined effect of P availability and post-fire drought. Our results provide insights on how plant functional traits modulate soil biochemical and microbiological response to fire in Mediterranean fire-prone shrublands.     

氮素对蕨麻克隆构型的可塑性反应

试验研究N素对蕨麻克隆构型的可塑性反应结果表明,蕨麻基株根长、分枝数、分株数、间隔子、地上干物质量、地下干物质量随N素的增加呈二次曲线变化,在施纯N 90.0kg/hm2时出现拐点,说明蕨麻对异质性资源(土壤N素)具有较强的可塑性。蕨麻总分枝长、根系膨大率、球根率则随N素水平的增加而直线下降,表明施用N素后蕨麻块根不具有商品价值。进行生态恢复时施用一定量N肥能显著提高蕨麻地上部分的生长,增大植被覆盖率。     

Arbuscular mycorrhizal fungi and endophytic fungi differentially modulate polyamines or proline of peach in response to soil flooding

Symbiotic fungi are involved in plant flooding tolerance, while the underlying mechanism is not yet known. Since polyamines (PAs) and proline are also associated with stress tolerance, it is hypothesized that the enhancement of stress resistance by symbiotic fungi is associated with changes in PAs and/or proline. The aim of this study was to analyze the effect of inoculation with Funneliformis mosseae and Serendipita indica on plant growth, PAs, and proline and the metabolisms in peach (Prunus persica) under flooding. Two-week flooding did not affect root colonization frequence of F. mosseae, while it promoted root colonization frequence of S. indica. Under flooding, plants inoculated with F. mosseae and S. indica maintained relatively higher growth rates than uninoculated plants. Funneliformis mosseae promoted root ornithine (Orn) contentration and arginine (Arg) and Orn decarboxylase activities under flooding, which promoted putrescine (Put), cadaverine (Cad), and spermidine (Spd) contentrations. Conversely, S. indica decreased contentrations of Arg, Orn, and agmatine and Arg decarboxylase activities, thus decreasing PA contentrations under flooding. Polyamines were negatively correlated with the expression of PA uptake transporter genes, PpPUT1 and PpPUT2, in peach. Polyamine transporter genes of F. mosseae (FmTPO) and S. indica (SiTPO) were regulated by flooding, of which FmTPO1 was positively correlated with Put, Cad, and Spd, along with positive correlations of Spd with SiTPO1, SiTPO2, and SiTPO4. Under flooding, F. mosseae decreased proline concentration, while S. indica increased proline concentration and correlated with expression of a △¹-pyrroline-5-carboxylate synthetase gene, PpP5CS2. It was thus concluded that F. mosseae modulated polyamine accumulation, while S. indica induced proline accumulation to tolerate flooding.     

Enhancement of plant essential oils' aqueous solubility and stability using alpha and beta cyclodextrin

Sodium benzoate has been shown to produce benzene in combination with ascorbic acid. This has led to research for safe alternatives from plant essential oils and parabens that have shown some antimicrobial activity, but many of these compounds exhibit poor solubility in aqueous solutions. Cyclodextrins can increase the solubility of many compounds. This work aimed to investigate the solubility of 23 plant essential oils and 4 parabens in water and an apple juice medium. Four of these compounds were chosen for their low aqueous solubility to determine if complexing the compound with α- and β-cyclodextrin would increase solubility. Three of the complexes were dissolved in an acidified aqueous solution and then studied in glass and polyethylene terephthalate (PET) to determine if storage material would affect the stability. Solubility of the 27 compounds in distilled water ranged from 1.6 mg/L to 2460.6 mg/L and the solubility of 18 of the compounds decreased from 2.5 to 84.7% in apple juice medium (pH = 3.4, 12-13 °Brix). Complexation with cyclodextrin dramatically increased the solubility of the compounds, up to 10-fold. Packaging material had no effect on concentration of compounds present over 7 days. Cyclodextrins were able to increase solubility of these compounds to more suitable concentrations, and may lead to viable natural alternatives to sodium benzoate.     

Isolation and identification of cucurbitane-type triterpenoids with partial agonist/antagonist potential for estrogen receptors from Momordica charantia

This study aims at investigating the estrogenic activity and active cucurbitane-type triterpenoid compounds of bitter gourd (Momordica charantia, MC) using a transactivation assay for estrogen receptors (ER) α and β. The lyophilized fruits of MC were exhaustively extracted with ethyl acetate (EA) and 95% ethanol (EtOH), sequentially. The nonsaponifiable fraction (NS) of the EA extract as well as the acid hydrolyzed EtOH extract (AH) was fractionated and isolated by repeated column chromatography and further purified by preparative HPLC or RP-HPLC. One known compound, 5β,19-epoxycucurbita-6,24-diene-3β,23ξ-diol (6), was isolated from the NS, and five new compounds (1-5) were isolated from AH and identified as cucurbita-6,22(E),24-trien-3β-ol-19,5β-olide (1), 5β,19-epoxycucurbita-6,22(E),24-triene-3β,19-diol (2), 3β-hydroxycucurbita-5(10),6,22(E),24-tetraen-19-al (3), 19-dimethoxycucurbita-5(10),6,22(E),24-tetraen-3β-ol (4), and 19-nor-cucurbita-5(10),6,8,22(E),24-pentaen-3β-ol (5). In the noncytotoxic concentration range, compounds 1, 2, 5 and 6 showed weak agonistic activity via ER α and β. Compounds 1, 2, 3 and 6 significantly antagonized the transactvation of 17β-estradiol (E(2)) via both ER α and β. In conclusion, this study demonstrates, for the first time as far as we know, the partial agonist/antagonist activity via ER of four new and one known cucurbitane-type triterpenoids from MC. Further studies are worthy to explore the selective estrogen receptor modulator (SERM) activity of MC.     

土壤和植物中元素含量的Zipf法则描述

土壤中含有人类已经发现的大多数化学元素,构成了植物矿质营养的主要来源.植物体内元素的种类也高达70余种,其含量从10-16到10-1,差异巨大.如果把土壤的化学组成和植物体内必须营养元素组成加以比较,可以看出两者之间存在相当大的差别.其中一些元素在土壤中的含量很高,但植物的需求量很低,如Si、Al、Fe等;而另一些元素在土壤中含量很低,但却是植物需求量较多的元素,如N、P、S;正是这些差异构成了土壤化学组成和植物营养之间关系的基础[1].元素的原子序数即核电荷数决定了元素的内在性质,一般的土壤和植物分析结果也表现出原子序数越大的重金属元素,其在土壤和植物中的含量也越低.Zipf法则由于具有普适性的特点,已被广泛地应用于离散型分布的研究[2],为了解土壤与植物中元素含量与元素原子序数之间存在的关系,利用Zipf法则对土壤与植物体内元素含量与原子序数之间的关系进行了研究.     

Landscape complexity differentially affects alpha, beta, and gamma diversities of plants occurring in fencerows and crop fields

Agro-ecosystems still retain part of the original biodiversity, although agricultural intensification threatens to eliminate refuge patches from farmland mosaics. Landscape complexity resulting from networks of uncultivated corridors associated with fencerows may play a key role in sustaining biodiversity across scales, and may further influence diversity in adjacent, cultivated fields. We evaluated the relationship between farmland complexity and plant diversity of fencerows and crop fields at local and landscape scales in the Rolling Pampas of Argentina. We surveyed 222 fencerows and fields cultivated with winter or summer crops, and characterised farmland complexity by the perimeter/area ratio of cropland in 2-km diameter circles surrounding each field. Plant diversity was additively partitioned into alpha, beta, and gamma components. Fencerows had noticeably higher richness than cropped fields at local and landscape scales. Gamma and beta diversities of fencerows and fields were positively related to farmland complexity, supporting the role of spatial heterogeneity in maintaining plant diversity in agro-ecosystems. Landscape complexity did not influence alpha diversity of fencerows but significantly increased diversity within fields, a likely result of enhanced mass effects from uncultivated habitats in more varied farmland. More complex landscapes contained greater gamma diversity of exotic perennials in fencerows, and of exotic and native annuals within fields. Importantly, alpha and gamma diversities of native perennials from the pristine Pampa grassland increased with landscape complexity within cropped fields. In the face of ongoing landscape homogenisation under agricultural intensification, maintaining fencerow networks may become critical for conserving habitat heterogeneity and farmland biodiversity.     

Predicting alpha, beta and gamma plant diversity from physiognomic and physical indicators as a tool for ecosystem monitoring

We searched for predictive models for alpha, beta and gamma plant diversity based in easy to measure field indicators. The study was conducted on the upper belt of the Córdoba mountains (Argentina). We established 222 permanent plots of 4 × 4 m distributed on sites with different physiognomy, topography and management. At each plot we measured physical and physiognomic indicators and recorded the presence of all vascular plants. We estimated alpha diversity as the number of species detected in a plot, beta diversity as the floristic dissimilarity between two plots, and gamma diversity as the number of species detected in a landscape. Through linear regression we found predictive models for alpha and pair-wise beta diversity. Then we analysed if predicted average alpha and beta diversity were good estimators of gamma diversity. We recorded a total of 288 species (5-74 species per plot). Alpha diversity was highest in sites on shallow soils with high structural richness (i.e. high number of cover categories), half covered by lawns, at sunny slopes and rough landscapes (r² = 0.66). For beta diversity, the difference between plots in structural richness and in cover of thick tussocks grasses and lawns were the best predictors (r² = 0.45). For different sets of simulated landscapes, gamma diversity was well explained by predicted average alpha and beta diversity, plus the sampling effort (r² = 0.92). We concluded that using easy to measure field indicators it is possible to estimate plant diversity at different levels with a good accuracy.     

Effects of coffee components on the response of GABA(A) receptors expressed in Xenopus oocytes

The effects of both coffee components and coffee extract on the electrical responses of GABA(A) receptors expressed in Xenopus oocytes were studied by injecting cRNAs of the alpha(1) and beta(1) subunits of the bovine receptors. The aqueous extract of coffee dose-dependently inhibited the GABA-elicited responses, whereas the lipophilic extract of coffee by diethyl ether slightly potentiated it at low doses (0.1-0.4 microL/mL) but showed inhibition at high doses (0.5-0.8 microL/mL). Theophylline inhibited the response in a noncompetitive mechanism (K(i) = 0.55 mM), whereas theobromine and trigonelline hydrochloride inhibited it in a competitive manner, K(i) = 3.8 and 13 mM, respectively. Benzothiazole, catechol, 2,4-dimethylstyrene, guaiacol, 1-octen-3-ol, sotolone, and 2,3,5-trimethylphenol potentiated the responses significantly. Potentiation elicited by guaiacol and sotolone was independent of GABA concentrations, whereas that by 1-octen-3-ol was dependent. When 1-octen-3-ol (100 mg/kg) was orally administered to mice prior to intraperitoneal administration of pentobarbital, the sleeping time of mice induced by pentobarbital increased significantly.     

Effects of tea components on the response of GABA(A) receptors expressed in Xenopus Oocytes

To study the effects of tea components on ionotropic gamma-aminobutyric acid (GABA) receptor response, ionotropic GABA receptors (GABA(A) receptors) were expressed in Xenopus oocytes by injecting cRNAs synthesized from cloned cDNAs of the alpha(1) and beta(1) subunits of the bovine receptors, and their electrical responses were measured by a voltage clamping method. Extracts of green tea, black tea, and oolong tea in an aqueous solution induced the GABA-elicited response, which showed that these teas contain GABA, whereas coffee does not. Caffeine weakly inhibited the response in a competitive manner (K(i) = 15 mM), and (+)-catechin inhibited it in a noncompetitive one (K(i) = 1.7 mM). Especially, two catechin derivatives, (-)-epicatechin gallate and (-)-epigallocatechin gallate, inhibited the response strongly. Alcohols such as leaf alcohol or linalool potentiated the response, possibly because their binding to the potentiation site enhances the GABA-binding affinity to GABA(A) receptors when they bind. Extracts of green tea made with ethyl ether, which must contain lipophilic components of green tea, inhibited the response elicited by GABA, possibly because the amounts of caffeine and catechin derivatives were much larger than fragrant alcohols in such extracts of tea.     

Cell-based and chemical assays of the ability to modulate the production of intracellular Reactive Oxygen Species of eleven Mediterranean plant species related to ethnobotanic traditions

The aim of this study was to compare three tests for the evaluation of the antioxidant potential of eleven plant species traditionally used to cure or to enhance animal and/or human wellbeing. In addition to the ability to modulate the production of intracellular reactive oxygen species the effect of the treatment on cell viability was also considered. In the cell-based experiment DCF (2′,7′dichlorofluorescein diacetate) and in the chemical assays DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2′-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt), Pistacia lentiscus exhibited high antioxidant capacity, while Cistus creticus and Euphorbia characias were ineffective in cell-culture and displayed high activity in the chemical assays, and others (Malva sylvestris, Matricaria chamomilla, Urtica dioica) showed an opposite trend. Olea europaea (sylvestris and europaea) and Umbilicus rupestris displayed highest antioxidant efficacy in both HUVEC and HL-60 cells, and provided also appreciable activity in the chemical assays. The study gives evidence that herbal extracts may have similar antioxidant properties but different effects on cells. Therefore, it is important to combine analytical and biological testing methods in order to measure the antioxidant capacity of natural extracts. Results encourage to explore further the potential nutraceutical and functional properties of Mediterranean plant species related to ethno-botanic traditions for animal health care.     

pH-dependent extraction of soil and soil amendments to understand the factors controlling element mobility

A combination of the pH-dependent leaching test CEN/TS 14429 and geochemical modelling (LeachXS^©-ORCHESTRA) was used to evaluate the partitioning of major, minor and trace elements in relation to their phosphorus retention capability, the release of non-nutrient constituents, their reduction of soil acidity and their organic matter retention. One Australian soil sample and two different soil amendments (Red Lime and Alkaloam) were studied. The pH-dependent leaching test showed that the acid neutralization capacity and the phosphorus retention of the soil were improved, respectively, by a factor of two to five and by a factor of two to ten after addition of these amendments. The amendments improved retention of dissolved organic matter (by a factor of 3–10) and did not increase leachability of undesired contaminants. The partitioning as obtained from modelling between free and dissolved organic-carbon-bound metals provides insight into the potential for uptake (bioavailability). This partitioning is very pH-dependent and therefore testing and associated chemical speciation modelling provide valuable information for judging traditional soil improvers and soil amendments, including the waste-derived soil amendments Alkaloam and Red Lime.     

Human IgE binding to the glycosidic moiety of bovine kappa-casein

IgE ability for recognizing milk proteins was assayed in the serum of an adult atopic patient who outgrew cow milk allergy in early childhood. A number of protein species included in casein from bovine milk were detected by human IgE in immunoblotting experiments. Comparing these results with those obtained from an analysis using antibody preparations specifically directed toward the different casein fractions, IgE-reactive bands were identified as isoforms of kappa-casein. IgE-reactive protein was not present in neither bovine cheese, regardless of cheese-making technology and time ripening, nor milk from any other dairy animal, such as ewe, goat, and water buffalo. Chemical deglycosylation of protein bands immobilized onto nitrocellulose proved that the glycosidic moiety of bovine kappa-casein was principally involved in IgE recognition.     

Covalent binding of the flavonoid quercetin to human serum albumin

Quercetin is an abundant flavonoid in the human diet with numerous biological activities, which may contribute to the prevention of human disease but also may be potentially harmful. Quercetin is oxidized in cells to products capable of covalently binding to cellular proteins, a process that may be important for its biological activities. In the present study, using radiolabeled drug and quantifying the products after electrophoretic separation, proteins to which oxidized quercetin is binding irreversibly were identified. The binding of quercetin to human serum albumin (HSA) in human blood and the effect of stimulation of neutrophilic myeloperoxidase on this binding were also measured. The in vitro binding of quercetin to eight proteins in the presence of catalytic amounts of horseradish peroxidase and hydrogen peroxide was highly selective for HSA. For all proteins the binding was dramatically decreased by reduced L-glutathione. In the blood samples, the release of neutrophilic myeloperoxidase by phorbol ester caused a 3-fold increase in the binding of quercetin to HSA. This study shows that quercetin in the presence of peroxidase/hydrogen peroxide covalently links to proteins with a particularly high affinity for HSA and that this also may occur in vivo after exposure to quercetin. This provides further insights into the complex behavior of this major dietary flavonoid.     

Modelling the response of terrestrial ecosystems to acidification and desiccation scenarios

Changes in vegetation are often caused by changes in abiotic site factors, such as pH, nitrogen availability and soil moisture. It has been recognized that abiotic site factors are affected by atmospheric deposition and groundwater-table changes. In order to evaluate the effects of eutrophication, acidification and desiccation on site factors, the model SMART2 has been developed. For the Netherlands combinations of two acidification and two seepage scenarios (1990–2050) were evaluated with SMART2. The results are focused on pH, nitrogen availability and base saturation. Calculations were made for combinations of five vegetation structures on seven soil types and the five groundwater-table classes, using a 1 km² grid. Results showed that deposition reductions lead to a relatively fast improvement of the site factors, increase in pH and base saturation and decrease in N availability. Whereas a reduction in groundwater abstractions of 25% has little or no effect on the pH and N availability.     

Studies of the binding of alpha-lactalbumin to immobilized peptide ligands.

The present work investigates the mechanism of binding of alpha-lactalbumin to the peptide ligand WHWRKR and its variants HWRKR and acetylated WHWRKR immobilized on a polymethacrylate chromatographic resin. The presence of two temperature-dependent binding mechanisms and one temperature-independent mechanism was demonstrated. Injections of different forms of alpha-lactalbumin (apo-alpha-lactalbumin, D87A mutant alpha-lactalbumin) displayed similar behaviors when compared to native alpha-lactalbumin, while lysozyme showed little or no binding to the WHWRKR and AcWHWRKR resins. An alternative process for isolation of alpha-lactalbumin from WPI was shown, using consecutive injections of WPI with limited elution.     

Increasing soy isoflavonoid content and diversity by simultaneous malting and challenging by a fungus to modulate estrogenicity

Soybeans were germinated on a kilogram-scale, by the application of malting technology used in the brewing industry, and concomitantly challenged with Rhizopus microsporus var. oryzae. In a time-course experiment, samples were taken every 24 h for 10 days, and the isoflavonoid profile was analyzed by RP-UHPLC-MS. Upon induction with R. microsporus, the isoflavonoid composition changed drastically with the formation of phytoalexins belonging to the subclasses of the pterocarpans and coumestans and by prenylation of the various isoflavonoids. The pterocarpan content stabilized at 2.24 mg of daidzein equivalents (DE) per g after ～9 days. The levels of the less common glyceofuran, glyceollin IV, and V/VI ranged from 0.18 to 0.35 mg DE/g and were comparable to those of the more commonly reported glyceollins I, II, and III (0.22-0.32 mg DE/g) and glycinol (0.42 mg DE/g). The content of prenylated isoflavones after the induction process was 0.30 mg DE/g. The total isoflavonoid content increased by a factor of 10-12 on DW basis after 9 days, which was suggested to be ascribable to de novo synthesis. These changes were accompanied by a gradual increase in agonistic activity of the extracts toward both the estrogen receptor α (ERα) and ERβ during the 10-day induction, with a more pronounced activity toward ERβ. Thus, the induction process yielded a completely different spectrum of isoflavonoids, with a much higher bioactivity toward the estrogen receptors. This, together with the over 10-fold increase in potential bioactives, offers promising perspectives for producing more, novel, and higher potency nutraceuticals by malting under stressed conditions.