Thiamine modulates metabolism of the phenylpropanoid pathway leading to enhanced resistance to Plasmopara viticola in grapevine.

BACKGROUND: Previously, we have reported the ability of thiamine (vitamin B1) to induce resistance against Plasmopara viticola in a susceptible grapevine cv. Chardonnay. However, mechanisms underlying vitamins, especially, thiamine-induced disease resistance in grapevine are still largely unknown. Here, we assessed whether thiamine could modulate phenylpropanoid pathway-derived phytoalexins in grapevine plants, as well as, the role of such secondary metabolites in thiamine-induced resistance process to P. viticola. RESULTS: Our data show that thiamine treatment elicited the expression of phenylpropanoid pathway genes in grapevine plants. The expression of these genes correlated with an accumulation of stilbenes, phenolic compounds, flavonoids and lignin. Furthermore, the total anti-oxidant potential of thiamine-treaded plants was increased by 3.5-fold higher level as compared with untreated-control plants. Four phenolic compounds are responsible of 97% of the total anti-oxidant potential of thiamine-treated plants. Among these compounds, is the caftaric acid, belonging to the hydroxy-cinnamic acids family. This element contributed, by its own, by 20% of this total anti-oxidant potential. Epifluorescence microscopy analysis revealed a concomitant presence of unbranched-altered P. viticola mycelia and stilbenes production in the leaf mesophyll of thiamine-treated inoculated plants, suggesting that stilbenes are an important component of thiamine-induced resistance in grapevine. CONCLUSION: This work is the first to show the role of thiamine, as a vitamin, in the modulation of grapevine plant secondary metabolism contributing to an enhanced resistance to P. viticola, the most destructive fungal disease in vineyards.

Effect of photocatalysis (TiO2/UVA) on the inactivation and inhibition of Pseudomonas aeruginosa virulence factors expression.

Water disinfection using visible light-active photocatalyst has recently attracted more attention due to its potential to inactivate microbes. In this study, we have investigated the efficiency of photocatalysis (TiO2/UVA) on the inactivation of Pseudomonas aeruginosa and the attenuation of its virulence factors. For this aim, the photocatalytic effects of TiO2/UVA on the cultivability and viability of P. aeruginosa were investigated. Furthermore, during the photocatalysis, the morphology of the bacterial cells was examined by atomic force microscopy (AFM) while the virulence factors were assessed by protease and lipase activities in addition to the mobility and communication of cells. The results revealed that during the photocatalysis the bacterial cells lost their cultivability and viability on agar under the action of the reactive oxygen species generated by the photocatalytic reaction. In addition, AFM observations have shown a damage of the bacterial membrane and a total disruption of the bacterial cells. Moreover, the major virulence factors such as biofilm, lipase and protease expression have been markedly inhibited by TiO2/UVA treatment. In addition, the bacteria lost their ability of communication 'quorum sensing' and mobility with twitching and swarming types after 60 min of photocatalytic treatment. Accordingly, TiO2/UVA is an effective method to reduce P. aeruginosa virulence and to prevent biofilm formation.

Comparative study on pilots between ANAMMOX favored conditions in a partially saturated vertical flow constructed wetland and a hybrid system for rural wastewater treatment.

The objective of this research was to evaluate the nitrogen removal in a single stage rural wastewater treatment system. It was a modified subsurface vertical flow (SSVF) constructed wetland. The so-called Anaerobic Ammonium Oxidation(ANAMMOX) process is favored by imposing a saturated zone at the bottom of the basin. The nitrogen removal performances of this modified SSVF were compared to those of a conventional hybrid system where the well-known nitrification-denitrification process is performed. This study was carried out using three lab-scale pilots of constructed wetlands during four months: (1) a hybrid constructed wetlands with a reed-Phragmites australis SSVF bed in serial with a cattail-Typha angustofolia SSHF bed (SSVFp + SSHF). (2) A reed-Phragmites australis SSVF bed partially saturated at 40% of its depth (SSVFPS); (3) A cattail-Typha angustofolia SSVF bed partially saturated at 40% of its depth (SSVFTS). The results showed that the three configurations used in this study were efficient for most of the pollutants reduction. In fact, single-stage reactors have achieved similar chemical oxygen demand (COD) removal in comparison to the two-stage reactor independently of the macrophytes species. However, for Total Kjeldahl Nitrogen (TKN), a slightly higher nitrogen removal efficiency was recorded for (SSVF p + SSHF) with an average removal rate of 53% versus 48% and 51% for SSVF PS and SSVFTS respectively. These findings were highlighted with fluorescent in situ hybridization (FISH) analysis, which demonstrated the presence of major differences in the community composition and abundance of the bacteria involved with denitrification and nitrification in the three systems. In fact, SSVFP of the hybrid system was characterized by highest relative abundance of nitrifying bacteria (13% Nitrosomonas, 11% Nitrosospira, 14% Nitrospira and 10% Nitrobacter). While, the SSHF of hybrid system had larger number of denitrifying species than SSVF, with relative abundances of pseudomonas (3%), Paracoccus (9%), Zoogloea (6%), Thauera (4%), Thiobacillus (2%) and Aeromonas (1%). Interestingly, in the SSVFST (planted with Thypha angustofolia) where the relative abundance of nitrifying bacteria was very low (4% Nitrosomonas, 4% Nitrosospira, 4% Nitrospira and 1% Nitrobacter), we have detected the presence of ANAMMOX bacteria (3%). Accordingly SSVFST in the presence of Thypha angustofolia have favored the development of ANAMMOX activity in comparison to the other configurations.

Effects of short- and long-term exposures of humic acid on the Anammox activity and microbial community.

Humic acid has a controversial effect on the biological treatment processes. Here, we have investigated humic acid effects on the Anammox activity by studying the nitrogen removal efficiencies in batch and continuous conditions and analyzing the microbial community using Fluorescence in situ hybridization (FISH) technique. The results showed that the Anammox activity was affected by the presence of humic acid at a concentration higher than 70 mg/L. In fact, in the presence of humic acid concentration of 200 mg/L, the Anammox activity decreased to 57% in batch and under continuous condition, the ammonium removal efficiencies of the reactor decreased from 78 to 41%. This reduction of Anammox activity after humic acid addition was highlighted by FISH analysis which revealed a considerable reduction of the abundance of Anammox bacteria and the bacteria living in symbiosis with them. Furthermore, a total inhibition of Candidatus Brocadia fulgida was observed. However, humic acid has promoted heterotrophic denitrifying bacteria which became dominant in the reactor. In fact, the evolution of the organic matter in the reactor showed that the added humic acid was used as carbon source by heterotrophic bacteria which explained the shift of metabolism to the favor of heterotrophic denitrifying bacteria. Accordingly, humic acid should be controlled in the influent to avoid Anammox activity inhibition.

Evaluation of a hybrid anaerobic biofilm reactor treating winery effluents and using grape stalks as biofilm carrier.

Wine production processes generate large amount of both winery wastewater and solid wastes. Furthermore, working periods, volumes and pollution loads greatly vary over the year. Therefore, it is recommended to develop a low-cost treatment technology for the treatment of winery effluents taking into account the variation of the organic loading rate (OLR). Accordingly, we have investigated the sequential operation of an anaerobic biofilm reactor treating winery effluents and using grape stalks (GSs) as biofilm carrier with an OLR ranging from 0.65 to 27 gCOD/L/d. The result showed that, during the start-up with wastewater influent, the chemical oxygen demand (COD) removal rate ranged from 83% to 93% and was about 91% at the end of the start-up period that lasted for 40 days. After 3 months of inactivity period of the reactor (no influent feeding), we have succeeded in restarting-up the reactor in only 15 days with a COD removal of 82% and a low concentration of volatile fatty acids (1 g/L), which confirms the robustness of the reactor. As a consequence, GSs can be used as an efficient carrier support, allowing a fast reactor start-up, while the biofilm conserves its activity during a non-feeding period. The proposed hybrid reactor thus permits to treat both winery effluents and GSs.

Sequential operation of a hybrid anaerobic reactor using a lignocellulosic biomass as biofilm support.

Agro-industries are facing many economic and environmental problems associated with seasonal generation of liquid and solid waste. In order to reduce treatment costs and to cope with seasonal variation, we have developed a hybrid anaerobic reactor operated sequentially by using lignocellulosic biomass (LB) as biofilm carrier support. Six LBs were tested to evaluate the treatment performance during a succession of two start-up periods, separated by a non-feeding period. After a short acclimation phase of several days, all the reactors succeeded in starting-up in less than 1month to reach an organic loading rate of 25gCODL(-1)d(-1). In addition, they restarted-up successfully in only 15days after a 3month non-feeding period, indicating that biofilms conserved their biological activities during this last phase. As a consequence, the use of LB as a biofilm support gives the potential to sustain seasonal variations of wastewater loads for industrial application.

Methionine elicits H2O2 generation and defense gene expression in grapevine and reduces Plasmopara viticola infection.

Methionine (Met) is a nutritionally essential sulfur-containing amino acid (SAA) known for its preponderant role as initiator in protein synthesis. However, other functions for Met in plants are not well described. The implication of this SAA in oxidative stress tolerance has been recently reported, however the mode of action of Met is still poorly understood. Here, we analyzed the elicitor activity of Met in grapevine as well as its effect on Plasmopara viticola resistance. The results show that Met induces hydrogen peroxide (H2O2) generation, a key element in plant defense signaling, and upregulates the expression of a battery of defense-related genes. Transcript levels of these genes were not further modulated by P. viticola inoculation of Met-pretreated plants, suggesting an elicitor role rather than a priming role for Met in grapevine. Met treatment also reduces P. viticola development in grapevine plants grown under glasshouse controlled-conditions. Fungitoxicity assays revealed that Met possesses a moderate antifungal activity compared with cysteine (Cys), another SAA known for its toxic effect to a large spectrum of fungi.

Characterization of ammonium retention processes onto cactus leaves fibers using FTIR, EDX and SEM analysis.

In order to reduce the impact of nitrogen pollution and to increase the agronomic value of plant wastes to be reused as organic fertilizer, we have investigated the removal of ammonium from aqueous solutions onto cactus leave fibers (CLF), and the mechanisms involved in the retention of ammonium at CLF surface. The results showed that ammonium retention onto these fibers occurred for a wide pH (6-10) and temperature ranges (20-60°C) and the biosorption potential of CLF increased with temperature from 1.4 to 2.3 mg g(-1) for initial concentration of 50 mg L(-1). The modeling studies showed that the ammonium biosorption was well described by the pseudo-second-order model, predicting therefore, chemisorption interactions-type at earlier stages and by intraparticle diffusion at later stages. Biosorption is governed by film diffusion process at higher concentrations and by particle diffusion process at higher temperatures. The surface of CLF determined by SEM revealed the presence of cracks and cavities which may allow the intraparticle diffusion and the ion exchange processes. Moreover, FTIR and EDX analysis before and after ammonium retention showed that the main mechanisms involved in the removal of ammonium were the ionic exchange by calcium ions as well as H(+) and the complexation with carboxylic, alcoholic and phenolic groups.

Thiamine induced resistance to Plasmopara viticola in grapevine and elicited host-defense responses, including HR like-cell death.

Recently, thiamine (VitaminB1) has been shown to induce resistance against Pseudomonas syringae in Arabidopsis plants through priming of defense responses. In this paper, we have demonstrated the efficiency of thiamine to induce resistance against downy mildew caused by the oomycete Plasmopara viticola in a susceptible Vitis vinifera cultivar "Chardonnay" under glasshouse controlled conditions by providing a dual mode of action involving direct antifungal activity and elicitation of host-defense responses. Thiamine-induced defense responses included the generation of hydrogen peroxide (H(2)O(2)) in both grapevine suspension cultured cells (SCC) and plant leaves, upregulation of an array of defense-related genes and the induction of other defense responses at subcellular level such as callose deposition in stomata cells, phenolic compounds accumulation and hypersensitive response (HR) like-cell death. Epifluorescence microscopy studies revealed dramatic changes in P. viticola individual developmental stages during its colonization of the intercellular space of the leaf mesophyll in thiamine-treated plants. Collectively, our report evidenced the efficiency of thiamine in the control of downy mildew in grapevine by direct and indirect effects, suggesting that thiamine could be an attractive alternative to chemical fungicides in disease management in vineyards.

Posidonia oceanica (L.) fibers as a potential low-cost adsorbent for the removal and recovery of orthophosphate.

Adsorption efficiency of orthophosphate from aqueous solution onto Posidonia oceanica fibers (POF) as a raw, natural and abundant material was investigated and compared with other common natural materials. A series of batch tests were undertaken to assess the effect of the system variables, i.e. initial aqueous orthophosphate concentration, contact time, adsorbent dosage, pH and temperature. Results indicate that orthophosphate uptake increased with increasing initial orthophosphate concentration, temperature and adsorbent dosage and decreased with increasing pH values. The maximum adsorption capacity (Q(m)) determined from the Langmuir isotherm was calculated to be 7.45 mg g(-1) for the studied orthophosphates concentration range of 15-100 mg L(-1), pH 7; adsorbent dosage of 2 g L(-1) and temperature of 20±2°C. The adsorption data were very well described by the pseudo-second order model predicting a chemisorption process. The energy dispersive spectroscopy (EDS) and FTIR analysis before and after adsorption of orthophosphate onto POF showed that the main involved mechanisms are ligand exchange between orthophosphate and Cl(-), SO(4)(2-) and OH(-) and precipitation with calcium. In comparison with other natural adsorbents, raw POF could be considered as one of the most efficient natural materials for the removal of orthophosphate with the possibility of agronomic reuse.

Removal of phosphorus from aqueous solution by Posidonia oceanica fibers using continuous stirring tank reactor.

The present study aims to develop a new potentially low-cost, sustainable treatment approach to soluble inorganic phosphorus removal from synthetic solutions and secondary wastewater effluents in which a plant waste (Posidonia oceanica fiber: POF) is used for further agronomic benefit. Dynamic flow tests using a continuous stirred tank reactor (CSTR) were carried out to study the effect of initial concentration of phosphorus, amount of adsorbent, feeding flow rate and anions competition. The experimental results showed that the removal efficiency of phosphorus from synthetic solutions is about 80% for 10 g L(-1) of POF. In addition, the variation of the initial concentration of phosphorus from 8 to 50 mg L(-1) increased the adsorption capacity from 0.99 to 3.03 mg g(-1). The use of secondary treated wastewater showed the presence of competition phenomenon between phosphorus and sulphate which could be overcoming with increasing the sorptive surface area and providing more adsorption sites when increasing the adsorbent dosage of POF. Compared with columns studies, this novel CSTR system showed more advantages for the removal of soluble phosphorus as a tertiary treatment of urban secondary effluents with more adsorption efficiency and capacity, in addition to the prospect use of saturated POF with nutriment as fertilizer and compost.

Ammonium biosorption onto sawdust: FTIR analysis, kinetics and adsorption isotherms modeling.

Sawdust, an available and renewable biomass, was investigated as a novel ammonium biosorbent. Biosorption occurred over a pH range of 6-10, reached an equilibrium state within only 20 min, and can be described by a pseudo-second-order model predicting a chemisorption process. Equilibrium data were very well represented by Langmuir isotherm and confirm monolayer coverage. FTIR analysis before and after biosorption of ammonium onto sawdust suggested that the main mechanisms involved in the removal of ammonium might be the ionic exchange and complexation. The use of sawdust presents an interesting option for both tertiary wastewater treatment (as a possible non-conventional biosorbent for the removal of ammonium), and waste recycling (as a fertilizer and compost).

Effect of temperature and pH on the biosorption of ammonium onto Posidonia oceanica fibers: equilibrium, and kinetic modeling studies.

In the present study, the effects of temperature and pH on ammonium biosorption onto Posidonia oceanica fibers were investigated. The results showed that the ammonium biosorption onto these fibers occurred for a wide pH range and the adsorption capacity of these fibers increased with increasing temperature. The modeling studies showed that the ammonium biosorption was well described by the pseudo-second-order model, predicting therefore chemisorption interactions-type at earlier stages and intraparticle diffusion at later stages. The ammonium biosorption was governed by film diffusion process at various temperatures. Besides, equilibrium data were very well represented by Langmuir isotherm, which confirmed the mono-layer coverage. The use of P. oceanica fibers presents an interesting option for both tertiary wastewater treatment (as a possible non-conventional biosorbent for the removal of ammonium), and waste recycling (as a fertilizer and compost).

Phosphate mine wastes reuse for phosphorus removal from aqueous solutions under dynamic conditions.

Phosphate mine slimes (PMS), an abundant waste generated from phosphate mines, was used in this study as a cost-effective adsorbent to investigate the phosphate anions removal from synthetic and urban secondary treated wastewater solutions. Dynamic experiments using laboratory reactors were carried out to study the effect of phosphate influent concentration, PMS dosage and feed flow rate on phosphate removal and a kinetic model was used to determine the phosphate mass transfer coefficients. The results show that the phosphate removal increases with influent phosphate concentration and PMS dosage. The feed flow rate has no significant effect. On the other hand, the phosphate removal from wastewater is less efficient than the synthetic solution due to anions competition process. The evaluation of phosphates mass transfer coefficients confirms the presence of anion competition phenomena and the necessity of increasing PMS dosage to provide more adsorption sites. The cost-effective and high adsorptive capability of PMS make them attractive materials for phosphate anions removal and recovery from secondary treated wastewaters with the possibility of agronomic reuse as fertilizer.