RESUMEN
To explore the mechanisms of crop straw degradation and phosphorus (P) release by phosphate-solubilizing fungi (PSF), a typical PSF Aspergillus niger (A. niger, ANG) was investigated for the degradation of wheat straw (WST) in this work. The results revealed that A. niger significantly increased wheat straw degradation (30%) compared with no A. niger treatment (7.7%). Meanwhile, more than 92% of total P was released from WST by A. niger, much higher than from WST treatment (69.5%). Although the ratios of inorganic P release between WST and WST + ANG treatments were similar (17.6 vs 19.7%), a significant difference occurred between their release of organic P, i.e., WST (51.9%) vs WST + ANG (72.5%). The high enzyme activity of ß-1,4-glucanase and ß-glucosidase produced by A. niger contributed to the wheat straw degradation and organic P release compared with no A. niger treatment. Oxalic acid secreted by A. niger dominated the release of inorganic P from WST. Our findings suggested that A. niger is an efficient microbial agent for crop straw degradation and P release, which could be a candidate in the pathway of straw return.
Asunto(s)
Aspergillus niger , Triticum , Aspergillus , Ácido Oxálico/metabolismo , Fosfatos/metabolismo , Fósforo/metabolismo , Triticum/metabolismoRESUMEN
Psammosilene tunicoides is a unique perennial medicinal plant species native to the Southwestern regions of China. Its wild population is rare and endangered due to over-excessive collection and extended growth (4-5 years). This research shows that H+-ATPase activity was a key factor for oxalate-inducing programmed cell death (PCD) of P. tunicoides suspension cells. Oxalic acid (OA) is an effective abiotic elicitor that enhances a plant cell's resistance to environmental stress. However, the role of OA in this process remains to be mechanistically unveiled. The present study evaluated the role of OA-induced cell death using an inverted fluorescence microscope after staining with Evans blue, FDA, PI, and Rd123. OA-stimulated changes in K+ and Ca2+ trans-membrane flows using a patch-clamp method, together with OA modulation of H+-ATPase activity, were further examined. OA treatment increased cell death rate in a dosage-and duration-dependent manner. OA significantly decreased the mitochondria activity and damaged its electron transport chain. The OA treatment also decreased intracellular pH, while the FC increased the pH value. Simultaneously, NH4Cl caused intracellular acidification. The OA treatment independently resulted in 90% and the FC led to 25% cell death rates. Consistently, the combined treatments caused a 31% cell death rate. Furthermore, treatment with EGTA caused a similar change in intracellular pH value to the La3+ and OA application. Combined results suggest that OA-caused cell death could be attributed to intracellular acidification and the involvement of OA in the influx of extracellular Ca2+, thereby leading to membrane depolarization. Here we explore the resistance mechanism of P. tunicoides cells against various stresses endowed by OA treatment.
Asunto(s)
Caryophyllaceae/metabolismo , Ácido Oxálico/metabolismo , Proteínas de Plantas/metabolismo , ATPasas de Translocación de Protón/metabolismo , Apoptosis , Caryophyllaceae/citología , Mitocondrias/metabolismo , Plantas Medicinales/citología , Plantas Medicinales/metabolismoRESUMEN
Sclerotinia sclerotiorum, a pathogen of more than 600 host plants, secretes oxalic acid to regulate the ambient acidity and provide conducive environment for pathogenicity and reproduction. Few Aspergillus spp. were previously proposed as potential biocontrol agents for S. sclerotiorum as they deteriorate sclerotia and prevent pathogen's overwintering and initial infections. We studied the nature of physical and biochemical interactions between Aspergillus and Sclerotinia. Aspergillus species inhibited sclerotial germination as they colonized its rind layer. However, Aspergillus-infested sclerotia remain solid and viable for vegetative and carpogenic germination, indicating that Aspergillus infestation is superficial. Aspergillus spp. of section Nigri (Aspergillus japonicus and Aspergillus niger) were also capable of suppressing sclerotial formation by S. sclerotiorum on agar plates. Their culture filtrate contained high levels of oxalic, citric and glutaric acids comparing to the other Aspergillus spp. tested. Exogenous supplementation of oxalic acid altered growth and reproduction of S. sclerotiorum at low concentrations. Inhibitory concentrations of oxalic acid displayed lower pH values comparing to their parallel concentrations of other organic acids. Thus, S. sclerotiorum growth and reproduction are sensitive to the ambient oxalic acid fluctuations and the environmental acidity. Together, Aspergillus species parasitize colonies of Sclerotinia and prevent sclerotial formation through their acidic secretions.
Asunto(s)
Ascomicetos/fisiología , Aspergillus/fisiología , Agentes de Control Biológico , Ácido Oxálico/metabolismo , Animales , Ascomicetos/crecimiento & desarrollo , Ascomicetos/metabolismo , Interacciones Microbianas , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , Esporas Fúngicas/crecimiento & desarrolloRESUMEN
Sclerotinia sclerotiorum (S. sclerotiorum) is a soil-borne pathogen with broad host range. Dissolved organic matter (DOM) plays a vital role in regulating microbial activity in soil. Exogenous selenium (Se) inhibits plant pathogen growth and enhances the capacity of plants to resist disease. DOM from rape straw with Se treated in soil (RSDOMSe) was extracted, and the inhibitory effect on S. sclerotiorum growth was investigated. RSDOMSe inhibited S. sclerotiorum growth, which not only caused severe damage to S. sclerotiorum hyphae but also enhanced soluble protein leakage, thereby improving the growth inhibition ratio by 20.9%. As the action in intercellular, RSDOMSe led to a significant increase in oxalic acid and decrease in CWDE (cell wall-degrading enzyme, which helps pathogens to invade plants) activities, downregulation of Bi1 (BAX inhibitor-1, required for S. sclerotiorum virulence), Ggt1 (γ-glutamyl transpeptidase, regulates the ROS antioxidant system), CWDE2 and CWDE10 gene expression levels, compared with non-Se treated RSDOM (RSDOMN). Eight metabolites upregulated in RSDOMSe were identified by GC-TOF-MS, and among these metabolites, fumaric acid, maleic acid, malonic acid, mucic acid, saccharic acid, succunic acid and phenylacetic acid showed significant inhibition on S. sclerotiorum growth. These findings provide valuable insight into a new approach for developing eco-friendly fungicides.
Asunto(s)
Ascomicetos/efectos de los fármacos , Brassica napus/química , Fungicidas Industriales/farmacología , Compuestos Orgánicos/farmacología , Selenio/química , Microbiología del Suelo , Suelo/química , Ascomicetos/crecimiento & desarrollo , Pared Celular/química , Fungicidas Industriales/química , Regulación Fúngica de la Expresión Génica/efectos de los fármacos , Hifa/efectos de los fármacos , Compuestos Orgánicos/química , Ácido Oxálico/metabolismo , Enfermedades de las Plantas/microbiologíaRESUMEN
Finding chromium-accumulating plants is of great interest for phytoremediation of soil contaminated by chromium (Cr). Inspired by Traditional Chinese Medicine, we examined the Cr-resistance and Cr-accumulation of Lonicera japonica Thunb. After a two-phase study using both soil and water culture, we found that L. japonica could be a novel Cr-accumulating plant, which contains an average Cr(III) content of 1297.14 mg.kg-1 in its leaves. The Cr enrichment factor and the Cr transport coefficient of Lonicera japonica was 5.19 and 1.79, respectively. Lonicera japonica is the fifth Cr-accumulating plant discovered worldwide, and the first Cr-accumulating woody plant ever discovered. The results support the conclusions drawn from studies of Cr-accumulating Leersia hexandra that oxalic acid production can increase Cr tolerance whereas citric acid or malic acid has no effect, suggesting that oxalic acid might be a common reason for Cr tolerance in all Cr-accumulating plants. Moreover, this study revealed that the production of anthocyanin and carotene can also increase Cr(III) tolerance, suggesting that anthocyanin and carotene might also account for Cr tolerance in Cr-accumulating plants. We believe that the discovery of Lonicera japonica as a Cr-accumulating plant will offer great opportunities in phytoremediation, and the success should be a strong sign that Traditional Chinese Medicine harbors more secrets to be uncovered with modern science.
Asunto(s)
Cromo/análisis , Lonicera/metabolismo , Contaminantes del Suelo/análisis , Biodegradación Ambiental , Carotenoides/metabolismo , Cromo/metabolismo , Lonicera/crecimiento & desarrollo , Medicina Tradicional China , Ácido Oxálico/metabolismo , Hojas de la Planta/metabolismo , Suelo/química , Contaminantes del Suelo/metabolismoRESUMEN
Basidiomycota fungi in the order Polyporales are specified to decomposition of dead wood and woody debris and thereby are crucial players in the degradation of organic matter and cycling of carbon in the forest ecosystems. Polyporales wood-decaying species comprise both white rot and brown rot fungi, based on their mode of wood decay. While the white rot fungi are able to attack and decompose all the lignocellulose biopolymers, the brown rot species mainly cause the destruction of wood polysaccharides, with minor modification of the lignin units. The biochemical mechanism of brown rot decay of wood is still unclear and has been proposed to include a combination of nonenzymatic oxidation reactions and carbohydrate-active enzymes. Therefore, a linking approach is needed to dissect the fungal brown rot processes. We studied the brown rot Polyporales species Fomitopsis pinicola by following mycelial growth and enzyme activity patterns and generating metabolites together with Fenton-promoting Fe3+-reducing activity for 3 months in submerged cultures supplemented with spruce wood. Enzyme activities to degrade hemicellulose, cellulose, proteins, and chitin were produced by three Finnish isolates of F. pinicola Substantial secretion of oxalic acid and a decrease in pH were notable. Aromatic compounds and metabolites were observed to accumulate in the fungal cultures, with some metabolites having Fe3+-reducing activity. Thus, F. pinicola demonstrates a pattern of strong mycelial growth leading to the active production of carbohydrate- and protein-active enzymes, together with the promotion of Fenton biochemistry. Our findings point to fungal species-level "fine-tuning" and variations in the biochemical reactions leading to the brown rot type of wood decay.IMPORTANCEFomitopsis pinicola is a common fungal species in boreal and temperate forests in the Northern Hemisphere encountered as a wood-colonizing saprotroph and tree pathogen, causing a severe brown rot type of wood degradation. However, its lignocellulose-decomposing mechanisms have remained undiscovered. Our approach was to explore both the enzymatic activities and nonenzymatic Fenton reaction-promoting activities (Fe3+ reduction and metabolite production) by cultivating three isolates of F. pinicola in wood-supplemented cultures. Our findings on the simultaneous production of versatile enzyme activities, including those of endoglucanase, xylanase, ß-glucosidase, chitinase, and acid peptidase, together with generation of low pH, accumulation of oxalic acid, and Fe3+-reducing metabolites, increase the variations of fungal brown rot decay mechanisms. Furthermore, these findings will aid us in revealing the wood decay proteomic, transcriptomic, and metabolic activities of this ecologically important forest fungal species.
Asunto(s)
Coriolaceae/metabolismo , Compuestos Férricos/metabolismo , Proteínas Fúngicas/metabolismo , Ácido Oxálico/metabolismo , Madera/microbiología , Coriolaceae/enzimología , Micelio/crecimiento & desarrollo , Oxidación-Reducción , PiceaRESUMEN
Experimental induction of hyperoxaluria by ethylene glycol (EG) administration is disapproved as it causes metabolic acidosis while the oral administration of chemically synthesized potassium oxalate (KOx) diet does not mimic our natural system. Since existing models comprise limitations, this study is aimed to develop an improved model for the induction of dietary hyperoxaluria, and nephrocalcinosis in experimental rats by administration of naturally available oxalate rich diet. Male albino Wistar rats were divided into five groups. Group I, control; group II rats received 0.75% EG, group III rats fed with 5% KOx diet and group IV and V rats were administered with spinach extract of 250 and 500 mg soluble oxalate/day respectively, for 28 d. Urine and serum biochemistry were analyzed. After the experimental period, rats were sacrificed, liver and kidney tissue homogenates were used for antioxidant and lipid peroxidation assay. Relative change in expression of kidney injury molecule-1 (KIM-1) and crystal modulators genes in kidney tissues were evaluated. Tissue damage was assessed by histology studies of liver and kidney. Experimental group rats developed hyperoxaluria and crystalluria. Urine parameters, serum biochemistry, antioxidant profile, lipid peroxidation levels and gene expression analysis of experimental group II and III rats reflected acute kidney damage compared to group V rats. Histopathology results showed moderate hyperplasia in liver and severe interstitial inflammation in kidneys of group II and III than group V rats. Ingestion of naturally available oxalate enriched spinach extract successfully induced dietary hyperoxaluria and nephrocalcinosis in rats with minimal kidney damage.
Asunto(s)
Modelos Animales de Enfermedad , Enfermedades Transmitidas por los Alimentos/etiología , Hiperoxaluria/etiología , Nefrocalcinosis/etiología , Ácido Oxálico/envenenamiento , Hojas de la Planta/efectos adversos , Spinacia oleracea/efectos adversos , Administración Oral , Animales , Biomarcadores/sangre , Biomarcadores/metabolismo , Biomarcadores/orina , Cristalización , Glicol de Etileno/toxicidad , Enfermedades Transmitidas por los Alimentos/metabolismo , Enfermedades Transmitidas por los Alimentos/patología , Enfermedades Transmitidas por los Alimentos/fisiopatología , Regulación de la Expresión Génica/efectos de los fármacos , Hiperoxaluria/metabolismo , Hiperoxaluria/patología , Hiperoxaluria/fisiopatología , Riñón/efectos de los fármacos , Riñón/metabolismo , Riñón/patología , Peroxidación de Lípido/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/metabolismo , Hígado/patología , Masculino , Nefrocalcinosis/metabolismo , Nefrocalcinosis/patología , Nefrocalcinosis/fisiopatología , Ácido Oxálico/administración & dosificación , Ácido Oxálico/química , Ácido Oxálico/metabolismo , Extractos Vegetales/efectos adversos , Extractos Vegetales/química , Hojas de la Planta/química , Ratas Wistar , Insuficiencia Renal/etiología , Spinacia oleracea/químicaRESUMEN
Fomitopsis pinicola is a species of Polyporales frequently encountered in Nordic temperate and boreal forests. In nature, the fungus causes destructive brown rot in wood, colonizing tree trunks often occupied by other Basidiomycota species. We mimicked these species-species interactions by introducing F. pinicola to five white rot species, all common saprotrophs of Norway spruce. Hyphal interactions and mycelial growth in various combinations were recorded, while activities of lignocellulose-acting CAZymes and oxidoreductases were followed in co-cultures on two different carbon-source media. Of the species, Phlebia radiata and Trichaptum abietinum were the strongest producers of lignin-modifying oxidoreductases (laccase, manganese peroxidase) when evaluated alone, as well as in co-cultures, on the two different growth media (low-nitrogen liquid medium containing ground coniferous wood, and malt extract broth). F. pinicola was an outstanding producer of oxalic acid (up to 61 mM), whereas presence of P. radiata prevented acidification of the growth environment in the liquid malt-extract cultures. When enzyme profiles of the species combinations were clustered, time-dependent changes were observed on wood-supplemented medium during the eight weeks of growth. End-point acidity and production of mycelium, oxalic acid and oxidoreductase activities, in turn clustered the fungal combinations into three distinct functional groups, determined by the presence of F. pinicola and P. radiata, by principal component analysis. Our findings indicate that combinations of wood-decay fungi have dramatic dynamic effects on the production of lignocellulose-active enzymes, which may lead to divergent degradative processes of dead wood and forest litter.
Asunto(s)
Basidiomycota/enzimología , Basidiomycota/crecimiento & desarrollo , Hifa/crecimiento & desarrollo , Tracheophyta/microbiología , Madera/microbiología , Basidiomycota/metabolismo , Ácido Oxálico/metabolismoRESUMEN
Phosphorus (P) availability in alkaline soils of arid and semi-arid regions is a major constraint for decreased crop productivity. Use of plant growth promoting rhizobacteria (PGPR) may enhance plant growth through the increased plant antioxidation activity. Additionally, PGPR may increase nutrient uptake by plants as a result of induced root exudation and rhizosphere acidification. The current study was aimed to investigate combined effects of P and Pesudomonas putida (PGPR) on chickpea growth with reference to antioxidative enzymatic activity and root exudation mediated plant nutrient uptake, particularly P. Half of the seeds were soaked in PGPR solution, whereas others in sterile water and latter sown in soils. Plants were harvested 8 weeks after onset of experiment and analyzed for leaf nutrient contents, antioxidant enzymes activities and organic acids concentrations. Without PGPR, P application (+P) increased various plant growth attributes, plant uptake of P and Ca, soil pH, citric acid and oxalic acid concentrations, whereas decreased the leaf POD enzymatic activity as compared to the P-deficiency. PGPR supply both under -P and +P improved the plant growth, plant uptake of N, P, and K, antioxidative activity of SOD and POD enzymes and concentrations of organic acids, whereas reduced the rhizosphere soil pH. Growth enhancement by PGPR supply was related to higher plant antioxidation activity as well as nutrient uptake of chickpea including P as a result of root exudation mediated rhizosphere acidification.
Asunto(s)
Cicer/crecimiento & desarrollo , Cicer/microbiología , Fósforo/farmacocinética , Pseudomonas putida/fisiología , Antioxidantes/metabolismo , Disponibilidad Biológica , Clorofila/metabolismo , Cicer/metabolismo , Ácido Cítrico/metabolismo , Enzimas/metabolismo , Concentración de Iones de Hidrógeno , Malatos/metabolismo , Ácido Oxálico/metabolismo , Fósforo/deficiencia , Exudados de Plantas/metabolismo , Hojas de la Planta/metabolismo , Rizosfera , Semillas/microbiología , Suelo/químicaRESUMEN
A local rice variety, "Baijiaolaojing", was grown in a paddy field in the Yuanyang rice terraces under ambient and supplemental levels of ultraviolet-B (UV-B, 280-315 nm) radiation. The effects of enhanced UV-B radiation (5 and 10 kJ m(-2) d(-1)) on methane emissions in the paddy field were evaluated using a closed-chamber gas chromatography-based system, and the contents of low-molecular-weight organic acids (LMWOAs) in root exudates were determined by high-performance liquid chromatography (HPLC). Peaks in methane emissions in the paddy field were detected at 60, 80 and 100 days after rice transplantation. The highest level of cumulative methane emissions occurred at the tillering stage, followed by the jointing-booting and maturity stages. The lowest level was found at the flowering stage. The enhanced UV-B radiation did not change the seasonal variation in methane emissions in the paddy field; however, it induced a significant increase in the flux of methane emissions at the jointing-booting and maturity stages, as well as a significant increase in the cumulative flux of methane emissions throughout the growth period. In addition, the enhanced UV-B radiation caused an increase in the contents of oxalic acid and succinic acid and a decrease in the contents of tartaric acid and malic acid in rice root exudates. Furthermore, a significant positive correlation (r = 0.725, p < 0.01) was found between the content of oxalic acid and the methane emissions in the paddy field. The results indicated that enhanced UV-B radiation promoted methane emissions in the paddy field, which was closely associated with its impact on the exudation of LMWOAs by rice roots.
Asunto(s)
Ácidos Carboxílicos/metabolismo , Metano/análisis , Oryza/metabolismo , Oryza/efectos de la radiación , Raíces de Plantas/metabolismo , Rayos Ultravioleta , Cromatografía de Gases , Cromatografía Líquida de Alta Presión , Malatos/metabolismo , Ácido Oxálico/metabolismo , Raíces de Plantas/efectos de la radiación , Estaciones del Año , Ácido Succínico/metabolismo , Luz Solar , Tartratos/metabolismo , Factores de TiempoRESUMEN
The effect of oxalic and itaconic acids, the inhibitors of the isocitrate lyase, on the production of isocitric acid by the wild strain Yarrowia lipolytica VKM Y-2373 grown in the medium containing rapeseed oil was studied. In the presence of oxalic and itaconic acids, strain Y. lipolytica accumulated in the medium isocitric acid (70.0 and 82.7 g/L, respectively) and citric acid (23.0 and 18.4 g/L, respectively). In control experiment, when the inhibitors were not added to the medium, the strain accumulated isocitric and citric acids at concentrations of 62.0 and 28.0 g/L, respectively. Thus, the use of the oxalic and itaconic acids as additives to the medium is a simple and convenient method of isocitric acid production with a minimum content of citric acid.
Asunto(s)
Isocitratos/metabolismo , Ácido Oxálico/farmacología , Aceites de Plantas/metabolismo , Succinatos/farmacología , Yarrowia/metabolismo , Ácido Cítrico/metabolismo , Inhibidores Enzimáticos/metabolismo , Inhibidores Enzimáticos/farmacología , Ácidos Grasos Monoinsaturados , Microbiología Industrial/métodos , Isocitratoliasa/antagonistas & inhibidores , Isocitratoliasa/metabolismo , Ácido Oxálico/metabolismo , Aceite de Brassica napus , Succinatos/metabolismo , Yarrowia/efectos de los fármacosRESUMEN
Tabebuia impetiginosa (Mart. ex DC.) Standl. has been used in traditional medicine for many centuries, being nowadays marketed as dried plant material (inner bark) for infusions, pills, and syrups. The main objective of the present work was to validate its popular use through the bioactivity evaluation of the inner bark (methanolic extract and infusion) and of two different formulations (pills and syrup) also based on the same plant-material. The antioxidant activity was evaluated by in vitro assays testing free radical scavenging activity, reducing power and inhibition of lipid peroxidation in brain homogenates. The cytotoxicity was determined in four human tumor cell lines (MCF-7, NCI-H460, HeLa and HepG2, and also in non-tumor cells (porcine liver primary cells, PLP2)). Furthermore, the sample was chemically characterized regarding free sugars, organic acids, fatty acids, and tocopherols. Syrup and methanolic extract showed the highest antioxidant activity, related to their highest amount of phenolics and flavonoids. Methanolic extract was the only sample showing cytotoxic effects on the tested human tumor cell lines, but none of the samples showed toxicity in PLP2. Glucose and oxalic acid were, respectively, the most abundant sugar and organic acid in the sample. Unsaturated predominated over the saturated fatty acids, due to oleic, linoleic, and linolenic acids expression. α- and γ-Tocopherols were also identified and quantified. Overall, T. impetiginosa might be used in different phytoformulations, taking advantage of its interesting bioactive properties and chemical composition.
Asunto(s)
Antineoplásicos Fitogénicos/farmacocinética , Antioxidantes/farmacología , Extractos Vegetales/farmacología , Tabebuia/química , Animales , Antineoplásicos Fitogénicos/química , Antioxidantes/química , Línea Celular , Línea Celular Tumoral , Suplementos Dietéticos , Flavonoides/química , Flavonoides/farmacología , Glucosa/metabolismo , Células HeLa , Células Hep G2 , Humanos , Peroxidación de Lípido/efectos de los fármacos , Células MCF-7 , Medicina Tradicional/métodos , Ácido Oxálico/metabolismo , Fenoles/química , Fenoles/farmacología , Extractos Vegetales/química , Porcinos , Tocoferoles/química , Tocoferoles/farmacologíaRESUMEN
Oxalate decarboxylase (OXDC) enzyme has immense biotechnological applications due to its ability to decompose anti-nutrient oxalic acid. Flammulina velutipes, an edible wood rotting fungus responds to oxalic acid by induction of OXDC to maintain steady levels of pH and oxalate anions outside the fungal hyphae. Here, we report that upon oxalic acid induction, a calmodulin (CaM) like protein-FvCaMLP, interacts with the OXDC promoter to regulate its expression. Electrophoretic mobility shift assay showed that FvCamlp specifically binds to two non-canonical E-box elements (AACGTG) in the OXDC promoter. Moreover, substitutions of amino acids in the EF hand motifs resulted in loss of DNA binding ability of FvCamlp. F. velutipes mycelia treated with synthetic siRNAs designed against FvCaMLP showed significant reduction in FvCaMLP as well as OXDC transcript pointing towards positive nature of the regulation. FvCaMLP is different from other known EF hand proteins. It shows sequence similarity to both CaMs and myosin regulatory light chain (Cdc4), but has properties typical of a calmodulin, like binding of (45)Ca(2+), heat stability and Ca(2+) dependent electrophoretic shift. Hence, FvCaMLP can be considered a new addition to the category of unconventional Ca(2+) binding transcriptional regulators.
Asunto(s)
Calmodulina/química , Carboxiliasas/química , Motivos EF Hand/genética , Flammulina/genética , Proteínas Fúngicas/química , Regulación Fúngica de la Expresión Génica , Secuencia de Aminoácidos , Secuencia de Bases , Sitios de Unión , Calcio/metabolismo , Calmodulina/genética , Calmodulina/metabolismo , Carboxiliasas/genética , Carboxiliasas/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Flammulina/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Hifa/genética , Hifa/metabolismo , Modelos Moleculares , Ácido Oxálico/metabolismo , Unión Proteica , Estructura Terciaria de Proteína , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Schizosaccharomyces/genética , Schizosaccharomyces/metabolismo , Alineación de SecuenciaRESUMEN
The devastating plant pathogen Sclerotinia sclerotiorum produces copious (up to 50 mM) amounts of oxalic acid, which, for over a quarter century, has been claimed as the pathogenicity determinant based on UV-induced mutants that concomitantly lost oxalate production and pathogenicity. Such a claim was made without fulfilling the molecular Koch's postulates because the UV mutants are genetically undefined and harbour a developmental defect in sclerotial production. Here, we generated oxalate-minus mutants of S. sclerotiorum using two independent mutagenesis techniques, and tested the resulting mutants for growth at different pHs and for pathogenicity on four host plants. The oxalate-minus mutants accumulated fumaric acid, produced functional sclerotia and have reduced ability to acidify the environment. The oxalate-minus mutants retained pathogenicity on plants, but their virulence varied depending on the pH and buffering capacity of host tissue. Acidifying the host tissue enhanced virulence of the oxalate-minus mutants, whereas supplementing with oxalate did not. These results suggest that it is low pH, not oxalic acid itself, that establishes the optimum conditions for growth, reproduction, pathogenicity and virulence expression of S. sclerotiorum. Exonerating oxalic acid as the primary pathogenicity determinant will stimulate research into identifying additional candidates as pathogenicity factors towards better understanding and managing Sclerotinia diseases.
Asunto(s)
Ascomicetos/genética , Ascomicetos/patogenicidad , Fumaratos/metabolismo , Ácido Oxálico/metabolismo , Enfermedades de las Plantas/microbiología , Ascomicetos/metabolismo , Concentración de Iones de Hidrógeno , Mutagénesis , Virulencia/genética , Factores de Virulencia/genéticaRESUMEN
This study evaluated the effect of alkaline industrial by-products such as flyash (FA) and redmud (RM) on phosphorus (P) mobilisation in abattoir wastewater irrigated soils, using incubation, leaching and plant growth (Napier grass [Pennisetum purpureum]) experiments. The soil outside the wastewater irrigated area was also collected and treated with inorganic (KH2PO4 [PP]) and organic (poultry manure [PM]) P treatments, to study the effect of FA and RM on P mobilisation using plant growth experiment. Among the amendments, FA showed the highest increase in Olsen P, oxalic acid content and phosphatase activity. The highest increase in Olsen P for PM treated non-irrigated soils showed the ability of FA and RM in mobilising organic P better than inorganic P (PP). There was over 85 % increase in oxalic acid content in the plant growth soils compared to the incubated soil, showing the effect of Napier grass in the exudation of oxalic acid. Both amendments (FA and RM) showed an increase in phosphatase activity at over 90 % at the end of the 5-week incubation period. The leaching experiment indicated a decrease in water soluble P thereby ensuring the role of FA and RM in minimising P loss to water bodies. FA and RM showed an increase in plant biomass for all treatments, where FA amended soil showed the highest increase as evident from FA's effect on Olsen P. Therefore, the use of FA and RM mobilised P in abattoir wastewater irrigated soils and increased biomass production of Napier grass plants through root exudation of oxalic acid.
Asunto(s)
Riego Agrícola , Ceniza del Carbón/química , Pennisetum/metabolismo , Fósforo/metabolismo , Aguas Residuales/química , Mataderos , Animales , Biomasa , Concentración de Iones de Hidrógeno , Residuos Industriales , Estiércol , Ácido Oxálico/metabolismo , Pennisetum/crecimiento & desarrollo , Monoéster Fosfórico Hidrolasas/metabolismo , Fósforo/química , Desarrollo de la Planta , Raíces de Plantas/fisiología , Aves de Corral , Suelo/química , ResiduosRESUMEN
Oxalate secretion was achieved in Pseudomonas fluorescens ATCC 13525 by incorporation of genes encoding Aspergillus niger oxaloacetate acetyl hydrolase (oah), Fomitopsis plaustris oxalate transporter (FpOAR) and Vitreoscilla hemoglobin (vgb) in various combinations. Pf (pKCN2) transformant containing oah alone accumulated 19 mM oxalic acid intracellularly but secreted 1.2 mM. However, in the presence of an artificial oxalate operon containing oah and FpOAR genes in plasmid pKCN4, Pf (pKCN4) secreted 13.6 mM oxalate in the medium while 3.6 mM remained inside. This transformant solubilized 509 µM of phosphorus from rock phosphate in alfisol which is 4.5 fold higher than the Pf (pKCN2) transformant. Genomic integrants of P. fluorescens (Pf int1 and Pf int2) containing artificial oxalate operon (plac-FpOAR-oah) and artificial oxalate gene cluster (plac-FpOAR-oah, vgb, egfp) secreted 4.8 mM and 5.4 mM oxalic acid, released 329 µM and 351 µM P, respectively, in alfisol. The integrants showed enhanced root colonization, improved growth and increased P content of Vigna radiata plants. This study demonstrates oxalic acid secretion in P. fluorescens by incorporation of an artificial operon constituted of genes for oxalate synthesis and transport, which imparts mineral phosphate solubilizing ability to the organism leading to enhanced growth and P content of V. radiata in alfisol soil.
Asunto(s)
Proteínas Bacterianas/genética , Hidrolasas/genética , Operón/genética , Ácido Oxálico/metabolismo , Fosfatos/metabolismo , Pseudomonas fluorescens , Hemoglobinas Truncadas/genética , Ácidos/farmacología , Aspergillus niger/genética , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Coriolaceae/genética , Hidrolasas/metabolismo , Hidrólisis , Organismos Modificados Genéticamente , Fósforo/metabolismo , Raíces de Plantas/metabolismo , Raíces de Plantas/microbiología , Pseudomonas fluorescens/genética , Pseudomonas fluorescens/metabolismoRESUMEN
During fungal rock phosphate (RP) solubilization, a significant quantity of fluoride (F(-)) is released together with phosphorus (P), strongly inhibiting the process. In the present study, the effect of two F(-) adsorbents [activated alumina (Al2O3) and biochar] on RP solubilization by Aspergillus niger was examined. Al2O3 adsorbed part of the F(-) released but also adsorbed soluble P, which makes it inappropriate for microbial RP solubilization systems. In contrast, biochar adsorbed only F(-) while enhancing phosphate solubilization 3-fold, leading to the accumulation of up to 160 mg of P per liter. By comparing the values of F(-) measured in solution at the end of incubation and those from a predictive model, it was estimated that up to 19 mg of F(-) per liter can be removed from solution by biochar when added at 3 g liter(-1) to the culture medium. Thus, biochar acted as an F(-) sink during RP solubilization and led to an F(-) concentration in solution that was less inhibitory to the process. In the presence of biochar, A. niger produced larger amounts of citric, gluconic, and oxalic acids, whether RP was present or not. Our results show that biochar enhances RP solubilization through two interrelated processes: partial removal of the released F(-) and increased organic acid production. Given the importance of organic acids for P solubilization and that most of the RPs contain high concentrations of F(-), the proposed solubilization system offers an important technological improvement for the microbial production of soluble P fertilizers from RP.
Asunto(s)
Aspergillus niger/metabolismo , Carbón Orgánico/química , Ácido Cítrico/metabolismo , Fluoruros/toxicidad , Sedimentos Geológicos/química , Gluconatos/metabolismo , Ácido Oxálico/metabolismo , Fósforo/metabolismo , Adsorción , Fluoruros/química , Fósforo/química , Suelo/química , Microbiología del Suelo , SolubilidadRESUMEN
Approximately 30% of the world's total land area and over 50% of the world's potential arable lands are acidic. Furthermore, the acidity of the soils is gradually increasing as a result of the environmental problems including some farming practices and acid rain. At mildly acidic or neutral soils, aluminium (Al) occurs primarily as insoluble deposits and is essentially biologically inactive. However, in many acidic soils throughout the tropics and subtropics, Al toxicity is a major factor limiting crop productivity. The Al-induced secretion of organic acid (OA) anions, mainly citrate, oxalate, and malate, from roots is the best documented mechanism of Al tolerance in higher plants. Increasing evidence shows that the Al-induced secretion of OA anions may be related to the following several factors, including (a) anion channels or transporters, (b) internal concentrations of OA anions in plant tissues, (d) temperature, (e) root plasma membrane (PM) H(+)-ATPase, (f) magnesium (Mg), and (e) phosphorus (P). Genetically modified plants and cells with higher Al tolerance by overexpressing genes for the secretion and the biosynthesis of OA anions have been obtained. In addition, some aspects needed to be further studied are also discussed.
Asunto(s)
Aluminio/farmacología , Ácido Cítrico/metabolismo , Malatos/metabolismo , Ácido Oxálico/metabolismo , Plantas/metabolismo , Adenosina Trifosfatasas/metabolismo , Aniones , Arabidopsis/efectos de los fármacos , Arabidopsis/metabolismo , Membrana Celular/enzimología , Productos Agrícolas/efectos de los fármacos , Productos Agrícolas/metabolismo , Regulación de la Expresión Génica de las Plantas , Magnesio/metabolismo , Nitroprusiato/farmacología , Fósforo/metabolismo , Proteínas de Plantas/metabolismo , Raíces de Plantas/enzimología , Plantas/efectos de los fármacos , Plantas Modificadas Genéticamente/efectos de los fármacos , Plantas Modificadas Genéticamente/metabolismo , Temperatura , Nicotiana/efectos de los fármacos , Nicotiana/metabolismoRESUMEN
Plants in association with soil microorganisms play an important role in mineral weathering. Studies have shown that plants in symbiosis with ectomycorrhizal (ECM) fungi have the potential to increase the uptake of mineral-derived nutrients. However, it is usually difficult to study many of the different factors that influence ectomycorrhizal weathering in a single experiment. In the present study, we carried out a pot experiment where Pinus patula seedlings were grown with or without ECM fungi in the presence of iron ore minerals. The ECM fungi used included Pisolithus tinctorius, Paxillus involutus, Laccaria bicolor and Suillus tomentosus. After 24 weeks, harvesting of the plants was carried out. The concentration of organic acids released into the soil, as well as potassium and phosphorus released from the iron ore were measured. The results suggest that different roles of ectomycorrhizal fungi in mineral weathering such as nutrient absorption and transfer, improving the health of plants and ensuring nutrient circulation in the ecosystem, are species specific, and both mycorrhizal roots and non-mycorrhizal roots can participate in the weathering process of iron ore minerals.
Asunto(s)
Compuestos de Hierro/metabolismo , Micorrizas/metabolismo , Pinus/microbiología , Microbiología del Suelo , Suelo/análisis , Transporte Biológico , Ácido Cítrico/análisis , Ácido Cítrico/metabolismo , Concentración de Iones de Hidrógeno , Hierro/metabolismo , Laccaria/crecimiento & desarrollo , Laccaria/metabolismo , Técnicas Microbiológicas/métodos , Micorrizas/crecimiento & desarrollo , Ácido Oxálico/análisis , Ácido Oxálico/metabolismo , Fósforo/metabolismo , Pinus/crecimiento & desarrollo , Pinus/metabolismo , Raíces de Plantas/metabolismo , Raíces de Plantas/microbiología , Potasio/metabolismo , Especificidad de la Especie , SimbiosisRESUMEN
INTRODUCTION: Boerhaavia diffusa Linn. (Nyctaginaceae) is widely used in traditional Indian medicines against renal afflictions including calcium oxalate (CaOx) urolithiasis and is known for antioxidant activity. OBJECTIVE: The present study was designed to investigate the ameliorating effect of aqueous extract of B. diffusa roots (BDE) in hyperoxaluric oxidative stress and renal cell injury. MATERIAL AND METHODS: In vitro antioxidant activity of BDE was estimated in terms of total phenolic content and 1,1-diphenyl-2-picryl hydrazyl free radical scavenging activity. Wistar albino rats were given 0.75% v/v ethylene glycol in drinking water to induce chronic hyperoxaluria and simultaneously BDE was given to nephrolithiasic treated rats at the dose of 100 and 200 mg/kg b.w. orally for 28 days. Urinary volume, oxalate, serum creatinine, blood urea nitrogen (BUN), malondialdehyde (MDA) and antioxidant enzyme (SOD, CAT, GST, GPx) were evaluated. RESULTS AND DISCUSSION: BDE extract was found to posses a high total phenolic content and exhibited significant free radicals scavenging activity. Oxalate excretion significantly increased in hyperoxaluric animals as compared to control which was protected in BDE-treated animals. BDE treatment significantly reduced level of MDA and improved the activity of antioxidant enzymes followed by reduction in BUN and serum creatinine. In addition, BDE reduced the number of CaOx monohydrate crystals in the urine. Histological analysis depicted that BDE treatment inhibited deposition of CaOx crystal and renal cell damage. CONCLUSION: The present study reveals that antioxidant activity of BDE significantly protects against hyperoxaluric oxidative stress and renal cell injury in urolithiasis.