Role of Jasmonic Acid Pathway in Tomato Plant-Pseudomonas syringae Interaction

The jasmonic acid pathway has been considered as the backbone of the response against necrotrophic pathogens. However, a hemi-biotrophic pathogen, such as Pseudomonas syringae, has taken advantage of the crosstalk between the different plant hormones in order to manipulate the responses for its own interest. Despite that, the way in which Pseudomonas syringae releases coronatine to activate jasmonic acid-derived responses and block the activation of salicylic acid-mediated responses is widely known. However, the implication of the jasmonic intermediates in the plant-Pseudomonas interaction is not studied yet. In this work, we analyzed the response of both, plant and bacteria using SiOPR3 tomato plants. Interestingly, SiOPR3 plants are more resistant to infection with Pseudomonas. The gene expression of bacteria showed that, in SiOPR3 plants, the activation of pathogenicity is repressed in comparison to wild type plants, suggesting that the jasmonic acid pathway might play a role in the pathogenicity of the bacteria. Moreover, treatments with JA restore the susceptibility as well as activate the expression of bacterial pathogenicity genes. The observed results suggest that a complete jasmonic acid pathway is necessary for the susceptibility of tomato plants to Pseudomonas syringae

Priming by Hexanoic Acid Induce Activation of Mevalonic and Linolenic Pathways and Promotes the Emission of Plant Volatiles

Hexanoic acid (Hx) is a short natural monocarboxylic acid present in some fruits and plants. Previous studies reported that soil drench application of this acid induces effective resistance in tomato plants against Botrytis cinerea and Pseudomonas syringae and in citrus against Alternaria alternata and Xanthomonas citri. In this work, we performed an in deep study of the metabolic changes produced in citrus by the application of Hx in response to the challenge pathogen A. alternata, focusing on the response of the plant. Moreover, we used 13C labeled hexanoic to analyze its behavior inside the plants. Finally, we studied the volatile emission of the treated plants after the challenge inoculation. Drench application of 13C labeled hexanoic demonstrated that this molecule stays in the roots and is not mobilized to the leaves, suggesting long distance induction of resistance. Moreover, the study of the metabolic profile showed an alteration of more than 200 molecules differentially induced by the application of the compound and the inoculation with the fungus. Bioinformatics analysis of data showed that most of these altered molecules could be related with the mevalonic and linolenic pathways suggesting the implication of these pathways in the induced resistance mediated by Hx. Finally, the application of this compound showed an enhancement of the emission of 17 volatile metabolites. Taken together, this study indicates that after the application of Hx this compound remains in the roots, provoking molecular changes that may trigger the defensive response in the rest of the plant mediated by changes in the mevalonic and linolenic pathways and enhancing the emission of volatile compounds, suggesting for the first time the implication of mevalonic pathway in response to hexanoic application.This work was supported by grants from “Programa de formació del personal investigador” of the Universitat Jaume I (PREDOC/2009/24), the Spanish Ministry of Science and Innovation AGL2010-22300-C03-01-02 and AGL2013-49023-C03-01-02-R co-funded by the European Regional Development Fund (ERDF)

Advances in endophytic fungi research: a data analysis of 25 years of achievements and challenges

Research on fungal endophytes has demonstrated the ability to improve crop performance and protect host plants against diverse biotic and abiotic stresses. Yet, despite the exponential growth of this topic, a whole outline to reflect the relevance and extent of each study type is missing. Hence, we performed an analysis of all available literature to expose the characteristics and limitations of this research field. Our results suggested that, overall, there is still a tendency to study the most known models in plant-fungal-stress combinations (ascomycetous fungi, grasses, abiotic stress). Fungal endophytes in dicot plants or against biotic stress, though promising, are still quite unexplored. All these data could lead future studies to assess less considered study factors that might help discern the beneficial effects of fungal endophytes with more extent and accuracy

NH4+ protects tomato plants against Pseudomonas syringae by activation of systemic acquired acclimation

NH4 + nutrition provokes mild toxicity by enhancing H2O2 accumulation, which acts as a signal activating systemic acquired acclimation (SAA). Until now, induced resistance mechanisms in response to an abiotic stimulus and related to SAA were only reported for exposure to a subsequent abiotic stress. Herein, the first evidence is provided that this acclimation to an abiotic stimulus induces resistance to later pathogen infection, since NH4 + nutrition (N-NH4 +)-induced resistance (NH4 +-IR) against Pseudomonas syringae pv tomato DC3000 (Pst) in tomato plants was demonstrated. N-NH4 + plants displayed basal H2O2, abscisic acid (ABA), and putrescine (Put) accumulation. H2O2 accumulation acted as a signal to induce ABA-dependent signalling pathways required to prevent NH4 + toxicity. This acclimatory event provoked an increase in resistance against later pathogen infection. N-NH4 + plants displayed basal stomatal closure produced by H2O2 derived from enhanced CuAO and rboh1 activity that may reduce the entry of bacteria into the mesophyll, diminishing the disease symptoms as well as strongly inducing the oxidative burst upon Pst infection, favouring NH4 +-IR. Experiments with inhibitors of Put accumulation and the ABA-deficient mutant flacca demonstrated that Put and ABA downstream signalling pathways are required to complete NH4 +-IR. The metabolic profile revealed that infected N-NH4 + plants showed greater ferulic acid accumulation compared with control plants. Although classical salicylic acid (SA)-dependent responses against biotrophic pathogens were not found, the important role of Put in the resistance of tomato against Pst was demonstrated. Moreover, this work revealed the cross-talk between abiotic stress acclimation (NH4 + nutrition) and resistance to subsequent Pst infection.This work was supported by grants from the Spanish Ministry of Science and Innovation (AGL2013-49023-C3-2-R). The authors are grateful to the Serveis Centrals d’Instrumentació Científica (SCIC) from Universitat Jaume I (UJI, Castellón, Spain)

Responsabilidad ambiental y futuro agrícola en las comarcas de Castellón

Material addicional de la ponència presentada al I Congrés Obert i Virtual Castelló 2020. Castelló de la Plana, 201

Combining remote sensing and GIS climate modelling to estimate daily forest evapotranspiration in a Mediterranean mountain area

Evapotranspiration monitoring allows us to assess the environmental stress on forest and agricultural ecosystems. Nowadays, Remote Sensing and Geographical Information Systems (GIS) are the main techniques used for calculating evapotranspiration at catchment and regional scales. In this study we present a methodology, based on the energy balance equation (B-method), that combines remote sensing imagery with GIS-based climate modelling to estimate daily evapotranspiration (ETd) for several dates between 2003 and 2005. The three main variables needed to compute ETd were obtained as follows: (i) Land surface temperature by means of the Landsat-5 TM and Landsat-7 ETM+ thermal band, (ii) air temperature by means of multiple regression analysis and spatial interpolation from meteorological ground stations data at satellite pass, and (iii) net radiation by means of the radiative balance. We calculated ETd using remote sensing data at different spatial and temporal scales (Landsat-7 ETM+, Landsat-5 TM and TERRA/AQUA MODIS, with a spatial resolution of 60, 120 and 1000 m, respectively) and combining three different approaches to calculate the B parameter, which represents an average bulk conductance for the daily-integrated sensible heat flux. We then compared these estimates with sap flow measurements from a Scots pine (Pinus sylvestris L.) stand in a Mediterranean mountain area. This procedure allowed us to better understand the limitations of ETd modelling and how it needs to be improved, especially in heterogeneous forest areas. The method using Landsat data resulted in a good agreement, R2 test of 0.89, with a mean RMSE value of about 0.6 mm day−1 and an estimation error of ±30 %. The poor agreement obtained using TERRA/AQUA MODIS, with a mean RMSE value of 1.8 and 2.4 mm day−1 and an estimation error of about ±57 and 50 %, respectively. This reveals that ETd retrieval from coarse resolution remote sensing data is troublesome in these heterogeneous areas, and therefore further research is necessary on this issue. Finally, implementing regional GIS-based climate models as inputs in ETd retrieval have has provided good results, making possible to compute ETd at regional scales

1-Methyltryptophan Treatment Increases Defense-Related Proteins in the Apoplast of Tomato Plants

The activation of induced resistance in plants may enhance the production of defensive proteins to avoid the invasion of pathogens. In this way, the composition of the apoplastic fluid could represent an important layer of defense that plants can modify to avoid the attack. In this study, we performed a proteomic study of the apoplastic fluid from plants treated with the resistance inducer 1-methyltryptophan (1-MT) as well as infected with Pseudomonas syringae pv. tomato (Pst). Our results showed that both the inoculation with Pst and the application of the inducer provoke changes in the proteomic composition in the apoplast enhancing the accumulation of proteins involved in plant defense. Finally, one of the identified proteins that are overaccumulated upon the treatment have been expressed in Escherichia coli and purified in order to test their antimicrobial effect. The result showed that the tested protein is able to reduce the growth of Pst in vitro. Taken together, in this work, we described the proteomic changes in the apoplast induced by the treatment and by the inoculation, as well as demonstrated that the proteins identified have a role in the plant protection

Resistance Induction in Olive Tree (Olea europaea) Against Verticillium Wilt by Two Beneficial Microorganisms and a Copper Phosphite Fertilizer

Enhancement of the natural defenses of a plant by beneficial microorganisms, i.e., endophytic bacteria and fungi or fertilizers such as copper phosphonates, could result in a potential alternative strategy against verticillium wilt of olive tree (Olea europaea). In this study, two beneficial microorganisms (the fungus Aureobasidium pullulans AP08 and the bacterium Bacillus amyloliquefaciens PAB-024) and a phosphonate salt copper phosphite (CuPh) were evaluated for their effectiveness as host resistance inducers against Verticillium dahliae in olive. To this end, 6-month-old healthy olive plants of the susceptible cultivar Picual were treated by foliar or root applications by spraying 15 ml per plant or by irrigation with 350 ml per plant of the dilutions of each product (CuPh: 3 or 10 ml l–1, respectively; PAB-024: 108 UFC ml–1; AP08: 106 UFC ml–1). Treatments were conducted weekly from 2 weeks before inoculation to 10 days after inoculation. A cornmeal–water–sand mixture (1:2:9; w:v:w) colonized by V. dahliae was used for plant inoculation. Additionally, treated and noninoculated, nontreated and inoculated, and nontreated and noninoculated plants were included for comparative purposes. Disease severity progress and shoot fresh weight were assessed. Parameters involved in plant resistance were monitored through determination and quantification of reactive oxygen species (ROS) response (H2O2), and evaluation of hormones was done by gene expression analysis. Aureobasidium pullulans and CuPh were the most effective in disease reduction in planta by foliar or root application, respectively. Plants treated with CuPh showed significantly higher shoot fresh weight compared to the other treatments. ROS was significantly enhanced in plants treated with B. amyloliquefaciens PAB-024 compared to the rest of treatments and control. With regard to the evaluation of hormones, high levels of salicylic acid were detected on leaves from all treatment combinations, but without significant enhancements compared to the nontreated control. Regarding the gene expression related to salicylic acid, only the WRKY5 gene has shown a strong enhancement in the treatment with B. amyloliquefaciens. On the other hand, a strong accumulation of jasmonic acid and jasmonic acid-isoleucine in plants treated with A. pullulans was observed in all the tissues analyzed and also in the roots of plants treated with B. amyloliquefaciens and CuPh

Putrescine Biosynthesis Inhibition in Tomato by DFMA and DFMO Treatment

This protocol can be used to inhibit the biosynthesis of polyamines, specifically putrescine, in tomato plants grown with NH4 + as a solely N source. In general, polyamines are positively charged small metabolites implicated in physiological processes, including organogenesis, embryogenesis, floral initiation and development, leaf senescence, pollen tube growth, fruit development and ripening and participate in the response to abiotic and biotic stresses (Tiburcio et al., 2014). Polyamines are synthesized from amino acids by decarboxylation of ornithine or arginine by ornithine decarboxylase (ODC) or arginine decarboxylase (ADC), respectively (Walters, 2003). Tomato plants grown with NH4 + as the sole N source presented an increase of putrescine content in leaves (Fernández-Crespo et al., 2015). To assess the importance of putrescine accumulation, DL-α-(Difluoromethyl)arginine (DFMA) and DL-α-(Difluoromethyl)ornithine (DFMO), inhibitors of putrescine synthesis, were used as irreversible inhibitors of ADC and ODC enzymes, respectively (Fallon and Phillips, 1988), with the purpose of reducing cellular putrescine accumulation induced by NH4 + nutrition. The inhibitor solution containing 2 mM DFMA and 5 mM DFMO was applied directly to each pot during the week prior to sample collection. Putrescine content was reduced by 35.3% in tomato plants grown with NH4 +.The work was supported by a grant from the Spanish Ministry of Science and Innovation (AGL2013- 49023-C-2-R

The Apoplast: A Key Player in Plant Survival

The apoplast comprises the intercellular space, the cell walls, and the xylem. Important functions for the plant, such as nutrient and water transport, cellulose synthesis, and the synthesis of molecules involved in plant defense against both biotic and abiotic stresses, take place in it. The most important molecules are ROS, antioxidants, proteins, and hormones. Even though only a small quantity of ROS is localized within the apoplast, apoplastic ROS have an important role in plant development and plant responses to various stress conditions. In the apoplast, like in the intracellular cell compartments, a specific set of antioxidants can be found that can detoxify the different types of ROS produced in it. These scavenging ROS components confer stress tolerance and avoid cellular damage. Moreover, the production and accumulation of proteins and peptides in the apoplast take place in response to various stresses. Hormones are also present in the apoplast where they perform important functions. In addition, the apoplast is also the space where microbe-associated molecular Patterns (MAMPs) are secreted by pathogens. In summary, the diversity of molecules found in the apoplast highlights its importance in the survival of plant cells