Campestris Infection Research Articles

Unravelling alternative splicing patterns in susceptible and resistant Brassica napus lines in response to Xanthomonas campestris infection

BackgroundRapeseed (Brassica napus L.) is an important oil and industrial crop worldwide. Black rot caused by the bacterial pathogen Xanthomonas campestris pv. campestris (Xcc) is an infectious vascular disease that leads to considerable yield losses in rapeseed. Resistance improvement through genetic breeding is an effective and sustainable approach to control black rot disease in B. napus. However, the molecular mechanisms underlying Brassica-Xcc interactions are not yet fully understood, especially regarding the impact of post-transcriptional gene regulation via alternative splicing (AS).ResultsIn this study, we compared the AS landscapes of a susceptible parental line and two mutagenized B. napus lines with contrasting levels of black rot resistance. Different types of AS events were identified in these B. napus lines at three time points upon Xcc infection, among which intron retention was the most common AS type. A total of 1,932 genes was found to show differential AS patterns between different B. napus lines. Multiple defense-related differential alternative splicing (DAS) hub candidates were pinpointed through an isoform-based co-expression network analysis, including genes involved in pathogen recognition, defense signalling, transcriptional regulation, and oxidation reduction.ConclusionThis study provides new insights into the potential effects of post-transcriptional regulation on immune responses in B. napus towards Xcc attack. These findings could be beneficial for the genetic improvement of B. napus to achieve durable black rot resistance in the future.

Novel pyrimidin-4-one derivatives as potential T3SS inhibitors against Xanthomonas campestris pv. campestris.

Cruciferous black rot is caused by Xanthomonas campestris pv. campestris (Xcc) infection and is a widespread disease worldwide. Excessive and repeated use of bactericide is an important cause of the development of bacterial resistance. It is imperative to take new approaches to screening compounds that target virulence factors rather than kill bacterial pathogens. The type III secretion system (T3SS) invades a variety of cells by transporting virulence effector factors into the cytoplasm and is an attractive antitoxic target. Toward the search of new T3SS inhibitors, an alternative series of novel pyrimidin-4-one derivatives were designed and synthesized and assessed for their effect in blocking the virulence. All of the target compounds were characterized by proton (1 H) nuclear magnetic resonance (NMR), carbon-13 (13 C) NMR, fluorine-19 (19 F) NMR and high-resolution mass spectrometry (HRMS). All compounds were evaluated using high-throughput screening systems against Xcc. The results of the biological activity test revealed that the compound SPF-9 could highly inhibit the activity of xopN gene promoter and the hypersensitivity (HR) of tobacco without affecting bacterial growth. Moreover, messenger RNA (mRNA) level measurements showed that compound SPF-9 inhibited the expression of some representative genes (hrp/hrc genes). Compound SPF-9 weakened the pathogenicity of Xcc to Raphanus sativus L. Compound SPF-9 has good potential for further development as a novel T3SS inhibitor against Xcc. © 2023 Society of Chemical Industry.

Comparative phylogenomic insights of KCS and ELO gene families in Brassica species indicate their role in seed development and stress responsiveness

Very long-chain fatty acids (VLCFAs) possess more than twenty carbon atoms and are the major components of seed storage oil, wax, and lipids. FAE (Fatty Acid Elongation) like genes take part in the biosynthesis of VLCFAs, growth regulation, and stress responses, and are further comprised of KCS (Ketoacyl-CoA synthase) and ELO (Elongation Defective Elongase) sub-gene families. The comparative genome-wide analysis and mode of evolution of KCS and ELO gene families have not been investigated in tetraploid Brassica carinata and its diploid progenitors. In this study, 53 KCS genes were identified in B. carinata compared to 32 and 33 KCS genes in B. nigra and B. oleracea respectively, which suggests that polyploidization might has impacted the fatty acid elongation process during Brassica evolution. Polyploidization has also increased the number of ELO genes in B. carinata (17) over its progenitors B. nigra (7) and B. oleracea (6). Based on comparative phylogenetics, KCS, and ELO proteins can be classified into eight and four major groups, respectively. The approximate date of divergence for duplicated KCS and ELO genes varied from 0.03 to 3.20 million years ago (MYA). Gene structure analysis indicated that the maximum number of genes were intron-less and remained conserved during evolution. The neutral type of selection seemed to be predominant in both KCS and ELO genes evolution. String-based protein-protein interaction analysis suggested that bZIP53, a transcription factor might be involved in the activation of transcription of ELO/KCS genes. The presence of biotic and abiotic stress-related cis-regulatory elements in the promoter region suggests that both KCS and ELO genes might also play their role in stress tolerance. The expression analysis of both gene family members reflect their preferential seed-specific expression, especially during the mature embryo development stage. Furthermore, some KCS and ELO genes were found to be specifically expressed under heat stress, phosphorus starvation, and Xanthomonas campestris infection. The current study provides a basis to understand the evolution of both KCS and ELO genes in fatty acid elongation and their role in stress tolerance.

Identification of Structural Features of Hydrocinnamic Acid Related to Its Allelopathic Activity against the Parasitic Weed Cuscuta campestris.

Cuscuta campestris is a parasitic weed species that inflicts worldwide noxious effects in many broadleaf crops due to its capacity to withdraw nutrients and water directly from the crop vascular system using haustorial connections. Cuscuta campestris control in the majority of crops affected is non-existent, and thus, research for the development of control methods is needed. Hydrocinnamic acid occurs naturally in the rhizosphere, playing regulatory roles in plant-plant and plant-microbe communities. The toxicity of hydrocinnamic acid against C. campestris was recently identified. In the present work, a structure-activity relationship study of 21 hydrocinnamic acid analogues was performed to identify key structural features needed for its allelopathic action against the seedling growth of this parasitic plant. The findings of this study provide the first step for the design of herbicides with enhanced activity for the control of C. campestris infection.

The Plant Defense Signal Salicylic Acid Activates the RpfB-Dependent Quorum Sensing Signal Turnover via Altering the Culture and Cytoplasmic pH in the Phytopathogen Xanthomonas campestris.

ABSTRACTPlant colonization by phytopathogens is a very complex process in which numerous factors are involved. Upon infection by phytopathogens, plants produce salicylic acid (SA) that triggers gene expression within the plant to counter the invading pathogens. The present study demonstrated that SA signal also directly acts on the quorum-sensing (QS) system of the invading pathogen Xanthomonas campestris pv. campestris to affect its virulence by inducing turnover of the diffusible signaling factor (DSF) family QS signal. First, Xanthomonas campestris pv. campestris infection induces SA biosynthesis in the cabbage host plant. SA cannot be degraded by Xanthomonas campestris pv. campestris during culturing. Exogenous addition of SA or endogenous production of SA induces DSF signal turnover during late growth phase of Xanthomonas campestris pv. campestris in XYS medium that mimics plant vascular environments. Further, the DSF turnover gene rpfB is required for SA induction of DSF turnover. However, SA does not affect the expression of rpfB and DSF biosynthesis gene rpfF at the transcriptional level. SA induction of DSF turnover only occurs under acidic conditions in XYS medium. Furthermore, addition of SA to XYS medium significantly increased both culture and cytoplasmic pH. Increased cytoplasmic pH induced DSF turnover in a rpfB-dependent manner. In vitro RpfB-dependent DSF turnover activity increased when pH increased from 6 to 8. SA exposure did not affect the RpfB-dependent DSF turnover in vitro. Finally, SA-treated Xanthomonas campestris pv. campestris strain exhibited enhanced virulence when inoculated on cabbage. These results provide new insight into the roles of SA in host plants and the molecular interactions between Xanthomonas campestris pv. campestris and cruciferous plants.

Interactive Regulation of Hormone and Resistance Gene in Proline Metabolism Is Involved in Effector-Triggered Immunity or Disease Susceptibility in the Xanthomonas campestris pv. campestris-Brassica napus Pathosystem.

To characterize cultivar variations in hormonal regulation of the transition between pattern-triggered immunity (PTI) and effector-triggered immunity or susceptibility (ETI or ETS), the responses of resistance (R-) genes, hydrogen peroxide, and proline metabolism in two Brassica napus cultivars to contrasting disease susceptibility (resistant cv. Capitol vs. susceptible cv. Mosa) were interpreted as being linked to those of endogenous hormonal levels and signaling genes based on a time course of disease symptom development. Disease symptoms caused by the Xanthomonas campestris pv. campestris (Xcc) infections were much more developed in cv. Mosa than in cv. Capitol, as shown by an earlier appearance (at 3 days postinoculation [3 DPI]) and larger V-shaped necrosis lesions (at 9–15 DPI) in cv. Mosa. The cultivar variations in the R-genes, hormone status, and proline metabolism were found in two different phases (early [0–3 DPI] and later [9–15 DPI]). In the early phase, Xcc significantly upregulated PTI-related cytoplasmic kinase (Botrytis-induced kinase-1 [BIK1]) expression (+6.3-fold) with salicylic acid (SA) accumulation in cv. Capitol, while relatively less (+2.6-fold) with highly increased jasmonic acid (JA) level in cv. Mosa. The Xcc-responsive proline accumulation in both cultivars was similar to upregulated expression of proline synthesis-related genes (P5CS2 and P5CR). During the later phase in cv. Capitol, Xcc-responsive upregulation of ZAR1 (a coiled-coil-nucleotide binding site-leucine-rich repeat [CC-NB-LRR-type R-gene]) was concomitant with a gradual increase in JA levels without additional proline accumulation. However, in cv. Mosa, upregulation of TAO1 (a toll/interleukin-1 receptor-nucleotide binding site-leucine-rich repeat [TIR-NB-LRR-type R-gene]) was consistent with an increase in SA and abscisic acid (ABA) levels and resulted in an antagonistic depression of JA, which led to a proline accumulation. These results indicate that Xcc-induced BIK1- and ZAR1-mediated JA signaling interactions provide resistance and confirm ETI, whereas BIK1- and TAO1-enhanced SA- and/or ABA-mediated proline accumulation is associated with disease susceptibility (ETS).

Search for Resistant Genotypes to Cuscuta campestris Infection in Two Legume Species, Vicia sativa and Vicia ervilia.

The dodders (Cuscuta spp.) are parasitic plants that feed on the stems of their host plants. Cuscuta campestris is one of the most damaging parasitic plants for the worldwide agricultural production of broad-leaved crops. Its control is limited or non-existent, therefore resistance breeding is the best alternative both economically and environmentally. Common vetch (Vicia sativa) and bitter vetch (Vicia ervilia) are highly susceptible to C. campestris, but no resistant genotypes have been identified. Thus, the aim of this study was to identify in V. sativa and V. ervilia germplasm collections genotypes resistant to C. campestris infection for use in combating this parasitic plant. Three greenhouse screening were conducted to: (1) identify resistant responses in a collection of 154 accessions of bitter vetch and a collection of 135 accessions of common vetch genotypes against infection of C. campestris; (2) confirm the resistant response identified in common vetch accessions; and (3) characterize the effect of C. campestris infection on biomass of V. sativa resistant and susceptible accessions. Most common vetch and bitter vetch genotypes tested were susceptible to C. campestris. However, the V. sativa genotype Vs.1 exhibited high resistance. The resistant phenotype was characterized by a delay in the development of C. campestris posthaustorial growth and a darkening resembling a hypersensitive-like response at the penetration site. The resistant mechanism was effective in limiting the growth of C. campestris as the ratio of parasite/host shoot dry biomass was more significantly reduced than the rest of the accessions. To the best or our knowledge, this is the first identification of Cuscuta resistance in V. sativa genotypes.

The Union Is Strength: The Synergic Action of Long Fatty Acids and a Bacteriophage against Xanthomonas campestris Biofilm.

Xanthomonas campestris pv. campestris is known as the causative agent of black rot disease, which attacks mainly crucifers, severely lowering their global productivity. One of the main virulence factors of this pathogen is its capability to penetrate and form biofilm structures in the xylem vessels. The discovery of novel approaches to crop disease management is urgent and a possible treatment could be aimed at the eradication of biofilm, although anti-biofilm approaches in agricultural microbiology are still rare. Considering the multifactorial nature of biofilm, an effective approach against Xanthomonas campestris implies the use of a multi-targeted or combinatorial strategy. In this paper, an anti-biofilm strategy based on the use of fatty acids and the bacteriophage (Xccφ1)-hydroxyapatite complex was optimized against Xanthomonas campestris mature biofilm. The synergic action of these elements was demonstrated and the efficient removal of Xanthomonas campestris mature biofilm was also proven in a flow cell system, making the proposed approach an effective solution to enhance plant survival in Xanthomonas campestris infections. Moreover, the molecular mechanisms responsible for the efficacy of the proposed treatment were explored.

Role of salicylic acid signaling in the biotrophy-necrotrophy transition of Xanthomonas campestris pv. campestris infection in Brassica napus

To characterize the hormonal regulations in the transition between biotrophic and necrotrophic phases of Xanthomonas campestris pv. campestris (Xcc) infection, the endogenous phytohormone and reactive oxygen species (ROS) levels and glutathione (GSH) redox status of non-inoculated and Xcc-infected Brassica napus cv. Mosa leaves were measured at various points after inoculation. This time-course study revealed two distinct phases of the Xcc-infection, namely an early asymptomatic (biotrophic) phase that could be observed at 2 days post-inoculation (DPI) and a destructive necrotrophic phase that occurred at 7–24 DPI and was associated with V-shaped necrosis. The length of the V-shaped lesions increased significantly with the progression of infection, as did H2O2 accumulation and the expression of NADPH oxidase. Xcc-infections induced the gradual depression of the reduced/oxidized GSH/GSSG ratio with the progression of disease. Furthermore, Xcc inoculation reduced levels of salicylic acid (SA), jasmonic acid (JA), abscisic acid (ABA), and cytokinin (CK) during the biotrophic phase (at 2 DPI); increased SA level (9.5 fold) and the expression of both SA synthesis-related genes (EDS1 and ICS1) and signaling-related genes (NPR1 and PR1) from the 2 to7 DPI. ABA and CK levels gradually increased during the necrotrophic phase. Interestingly, JA level was either reduced or not significantly affected during the necrotrophic phase of infection. Altogether, these results indicate that, in B. napus, changes in SA-mediated ROS production regulate the transition from the symptomless biotrophic phase of Xcc infection to the destructive necrotrophic phase.

Genome-Wide Identification, Expression Profile of the TIFY Gene Family in Brassica oleracea var. capitata, and Their Divergent Response to Various Pathogen Infections and Phytohormone Treatments.

TIFY, a plant-specific gene family with the conserved motif TIF[F/Y]XG, plays important roles in various plant biological processes. Here, a total of 36 TIFY genes were identified in the Brassica oleracea genome and classified into JAZ (22 genes), TIFY (7 genes), ZML (5 genes), and PPD (2 genes) subfamilies based on their conserved motifs, which were distributed unevenly across nine chromosomes with different lengths (339–1077 bp) and exon numbers (1–8). Following phylogenetic analysis with A. thaliana and B. rapa TIFY proteins, ten clades were obtained. The expression of these TIFY genes was organ-specific, with thirteen JAZ genes and two PPD genes showing the highest expression in roots and leaves, respectively. More importantly, the JAZs showed divergent responses to various pathogen infections and different phytohormone treatments. Compared with the susceptible line, most JAZs were activated after Plasmodiophora brassicae infection, while there were both induced and inhibited JAZs after Fusarium oxysporum or Xanthomonas campestris infection in the resistance line, indicating their probably distinct roles in disease resistance or susceptibility. Further, the JAZs were all upregulated after MeJA treatment, but were mostly downregulated after SA/ET treatment. In summary, these results contribute to our understanding of the TIFY gene family, revealing that JAZs may play crucial and divergent roles in phytohormone crosstalk and plant defense.

Calcium-signaling proteins mediate the plant transcriptomic response during a well-established Xanthomonas campestris pv. campestris infection

The plant immune system is divided into two branches; one branch is based on the recognition of pathogen-associated molecular patterns (PAMP-triggered immunity), and the other relies on pathogenic effector detection (effector-triggered immunity). Despite each branch being involved in different complex mechanisms, both lead to transcription reprogramming and, thus, changes in plant metabolism. To study the defense mechanisms involved in the Brassica oleracea–Xanthomonas campestris pv. campestris (Xcc) interaction, we analyzed the plant transcriptome dynamics at 3 and 12 days postinoculation (dpi) by using massive analysis of 3′-cDNA ends. We identified more induced than repressed transcripts at both 3 and 12 dpi, although the response was greater at 12 dpi. Changes in the expression of genes related to the early infection stages were only detected at 12 dpi, suggesting that the timing of triggered defenses is crucial to plant survival. qPCR analyses in susceptible and resistant plants allowed us to highlight the potential role of two calcium-signaling proteins, CBP60g and SARD1, in the resistance against Xcc. This role was subsequently confirmed using Arabidopsis knockout mutants.

BDSF Is the Predominant In-Planta Quorum-Sensing Signal Used During Xanthomonas campestris Infection and Pathogenesis in Chinese Cabbage.

Xanthomonas campestris pv. campestris uses the diffusible signal factor (DSF) family of quorum-sensing (QS) signals to coordinate virulence and adaptation. DSF family signals have been well-characterized using laboratory-based cell cultures. The in-planta QS signal used during X. campestris pv. campestris infection remains unclear. To achieve this goal, we first mimic in-planta X. campestris pv. campestris growth conditions by supplementing the previously developed XYS medium with cabbage hydrolysate and found that the dominant signal produced in these conditions was BDSF. Secondly, by using XYS medium supplemented with diverse plant-derived compounds, we examined the effects of diverse plant-derived compounds on the biosynthesis of DSF family signals. Several compounds were found to promote biosynthesis of BDSF. Finally, using an X. campestris pv. campestris ΔrpfB-Chinese cabbage infection model and an ultra-performance liquid chromatographic-time of flight-mass spectrometry-based assay, BDSF was found to comprise >70% of the DSF family signals present in infected cabbage tissue. BDSF at a concentration of 2.0 μM induced both protease activity and engXCA expression. This is the first report to directly show that BDSF is the predominant in-planta QS signal used during X. campestris pv. campestris infection. It provides a better understanding of the molecular interactions between X. campestris pv. campestris and its cruciferous hosts and also provides the logical target for designing strategies to counteract BDSF signaling and, thus, infection. Further studies are needed to get an exact idea about the DSF production dynamics of the wild-type strain inside the plant.

Evaluation of the potential yield and primary symptoms of Xanthomonas campestris pv. campestris infection in Asian vegetables grown in the Czech Republic

Selected cultivars of Asian brassicacean vegetables were evaluated for their yield potential and susceptibility to bacterial infection. Chinese broccoli, two cultivars of Chinese cabbage (‘Dwarf milk cabbage’ and improved ‘Tahtsai’), and mizuna were grown in the conditions of the Czech Republic. Morphological and yield parameters for the field conditions (plant height, plant diameter, and marketable weight) were verified. In addition, genotypes were tested for resistance to black rot [<em>Xanthomonas campestris</em> pv. <em>campestris</em> (Xcc)] at the planting stage. The results show that a very promising genotype of mizuna, which yielded best (1,478 g per plant). Chinese cabbage (cultivar ‘Dwarf milk cabbage’) also showed high yield values (2,839 g per plant), especially when compared to the reference value for Napa cabbage. Chinese Cabbage 1 (‘Dwarf milk cabbage’) and mizuna also showed a low susceptibility to infection by bacterial black rot at the planting stage. Both cultivars reached Level 2 (median) of infection, which corresponds to the extent of the symptoms on 25% of the leaf surface.

Salinity effect on Cuscuta campestris Yunck. Parasitism on Arabidopsis thaliana L.

Stem holoparasitic flowering plants of the genus Cuscuta are globally distributed invasive species and agricultural pests. The present research represents the combined effect of salt stress (e.g. abiotic stress) and Cuscuta campestris infection (e.g. biotic stress) on the model host plant Arabidopsis thaliana and the response of the parasite to salinity. The response of these parasites to abiotic stress conditions including salinity is poorly studied. Arabidopsis plants were continuously irrigated with 0, 50 and 150 mM NaCl and subjected to C. campestris infection. The influence of both abiotic and biotic stresses on the major osmoprotectant L-proline and three antioxidant enzymes – catalase, superoxide dismutase and guaiacol peroxidase, was assessed in both the parasite and the host plant. All four biochemical markers were differentially affected by stress, showing that the influence of C. campestris parasitism and its interaction with salinity is mostly in the site of infection (direct response) and also in roots (indirect vertical response) rather than on non-infected leaves of infected plants (indirect horizontal response). Despite its absence of soil contact, C. campestris was also significantly affected by salinity (indirect response). The mutual adaptation of the parasite-host pair to salinity slightly altered the regular response to abiotic stress of A. thaliana, but no detrimental additive effect of biotic and abiotic stress was observed.

Unraveling the metabolic response of Brassica oleracea exposed to Xanthomonas campestris pv. campestris.

Brassica crops together with cereals represent the basis of world supplies. Due to their importance, the production losses caused by Xanthomonas campestris pv. campestris (Xcc) infection represent a high economic impact. Understanding molecular and biochemical mechanisms of plants is essential to develop resistant crops with durable protection against diseases. In this regard, metabolomics has emerged as a valuable technology to provide an overview of the biological status of a plant exposed to a disease. This study investigated the dynamic changes in the metabolic profile of Brassica oleracea plants during an Xcc infection from leaves collected at five different days post infection using a mass spectrometry approach. Results showed that Xcc infection causes dynamic changes in the metabolome of B. oleracea. Moreover, induction/repression pattern of the metabolites implicated in the response follows a complex dynamics during infection progression, indicating a complex temporal response. Specific metabolic pathways such as alkaloids, coumarins or sphingolipids are postulated as promising key role candidates in the infection response. This work tries to decipher the changes produced on Brassica crops metabolome under Xcc infection and represents a step forward in the understanding of B. oleracea-Xcc interaction. © 2018 Society of Chemical Industry.

Loss of chloroplast-localized protein phosphatase 2Cs in Arabidopsis thaliana leads to enhancement of plant immunity and resistance to Xanthomonas campestris pv. campestris infection.

Protein phosphatases (PPs) counteract kinases in reversible phosphorylation events during numerous signal transduction pathways in eukaryotes. PP2Cs, one of the four major classes of the serine/threonine-specific PP family, are greatly expanded in plants. Thus, PP2Cs are thought to play a specific role in signal transduction pathways. Some rice PP2Cs classified in subgroup K are responsive to infection by the compatible Xanthomonas oryzae pv. oryzae, the causal agent of bacterial blight. In Arabidopsis thaliana, orthologous PP2C genes (AtPP2C62 and AtPP2C26) classified to subgroup K are also responsive to Xanthomonas campestris pv. campestris (Xcc, causal agent of black rot) infection. To elucidate the function of these subgroup K PP2Cs, atpp2c62- and atpp2c26-deficient A.thaliana mutants were characterized. A double mutant plant which was inoculated with a compatible Xcc showed reduced lesion development, as well as the suppression of bacterial multiplication. AtPP2C62 and AtPP2C26 localized to the chloroplast. Furthermore, the photosynthesis-related protein, chaperonin-60, was indicated as the potential candidate for the dephosphorylated substrate catalysed by AtPP2C62 and AtPP2C26 using two-dimensional isoelectric focusing sodium dodecylsulfate-polyacrylamide gel electrophoresis (2D-IDF-SDS-PAGE). Taken together, AtPP2C62 and AtPP2C26 are suggested to be involved in both photosynthesis and suppression of the plant immune system. These results imply the occurrence of crosstalk between photosynthesis and the plant defence system to control productivity under pathogen infection.

Stimulation effect of carrageenan on enzymatic defense system of sweet basil against Cuscuta campestris infection

ABSTRACTSweet basil is an important medicinal plant used especially for therapeutical potentials. Sweet basil is a common host for Cuscuta campestris, which has a negative effect on infected plants. Therefore, natural friendly control of C. campestris seems to be useful. It has been shown that carrageenans can act as elicitors of plant defense responses. In this work, the effect of κ-carrageenans on protection against C. campestris and suppression of its invasion in basils were studied. Basils were sprayed with a solution of κ-carrageenan (1 g L−1), once a week, three times in total. Infection of basils with C. campestris was performed two days after the last carrageenan treatment. C. campestris stem and the leaves of basils were collected two weeks after C. campestris inoculation for biochemical studies. Treatment with carrageenan significantly increased shoot length and leaf area of basil and decreased C. campestris infestation by about 26%. The content of malondialdehyde, other aldehydes, hydrogen peroxide and lipoxygenase (LOX) activity increased significantly in basils parasitized by C. campestris. There were significant differences in phenylalanine ammonia lyase (PAL), catalase (CAT), superoxide dismutase (SOD) and peroxidase activity of parasitized basils by C. campestris compared with healthy basils. Carrageenan treatment of basils caused a significant increase in H2O2 content and the activity of PAL, CAT and SOD, but not of malondialdehyde, other aldehydes content and LOX, polyphenol oxidase (PPO) and peroxidases activity. The activated enzymatic defense system (PAL, PPO, CAT, SOD and peroxidase) in carrageenan-treated basils have a vital role in alleviating oxidative stress damage in infected plants, by removing excess reactive oxygen species and inhibiting LOX activity and lipid peroxidation that was observed in this study. Our results showed that the application of κ-carrageenan-induced beneficial effects in plants, with regard to growth stimulation and the activation of enzymatic defense system. Thus, carrageenan was recommended as a natural biostimulator for protection of plants against C. campestris.

Priming of pathogenesis related-proteins and enzymes related to oxidative stress by plant growth promoting rhizobacteria on rice plants upon abiotic and biotic stress challenge

Two plant growth promoting rhizobacteria (PGPR) were tested to evaluate their capacity to prime rice seedlings against stress challenge (salt and Xanthomonas campestris infection). As is accepted that plants respond to biotic and abiotic stresses by generation of reactive oxygen species (ROS), enzyme activities related to oxidative stress (ascorbate peroxidase (APX, EC 1.11.1.11), guaiacol peroxidase (GPX, EC 1.11.1.7), glutathione reductase (GR, EC 1.6.4.2) and superoxide dismutase (SOD, EC 1.15.1.1)) as well as the pathogenesis-related proteins (PRs) ß-1,3-glucanase (PR2, EC 3.2.1.6) and chitinase (PR3, EC 3.2.1.14) were measured at 3 time points after stress challenge. In addition, photosynthetic parameters related with fluorescence emission of photosystem II (F0, Fv/Fm, ΦPSII and NPQ) were also measured although they were barely affected. Both strains were able to protect rice seedlings against salt stress. AMG272 reduced the salt symptoms over 47% with regard to control, and L81 over 90%. Upon pathogen challenge, 90% protection was achieved by both strains. All enzyme activities related to oxidative stress were modified by the two PGPR, especially APX and SOD upon salinity stress challenge, and APX and GR upon pathogen presence. Both bacteria induced chitinase activity 24 and 48h after pathogen inoculation, and L81 induced ß-1,3-Glucanase activity 48h after pathogen inoculation, evidencing the priming effect. These results indicate that these strains could be used as bio-fortifying agents in biotechnological inoculants in order to reduce the effects of different stresses, and indirectly reduce the use of agrochemicals.

Beneficial rhizobacteria from rice rhizosphere confers high protection against biotic and abiotic stress inducing systemic resistance in rice seedlings

The present study reports a screening for PGPR in a highly selective environment, the rhizosphere of rice plants, in southwestern of Spain. Among the 900 isolates, only 38% were positive for at least one of the biochemical activities to detect putative PGPR. The best 80 isolates were selected and identified by 16S rRNA partial sequencing. Among these, 13 strains were selected for growth promotion assays. Only one strain (BaC1-38) was able to significantly increase height, while nine strains significantly inhibited it. Five strains significantly increased dry weight, and only BaC1-21 significantly decreased it. Based on significant modifications in growth, three bacteria (BaC1-13, BaC1-21 and BaC1-38) were tested for systemic induction of resistance against stress challenge (salt and Xanthomonas campestris infection). Protection against salt stress and pathogen infection was similar; BaC1-38 protected by 80%, BaC1-13 by 50% and BaC1-21 only by 20%. Toxicity of salt stress to the plants was evaluated by photosynthetic efficiency of seedlings. Fv/Fm only decreased significantly in plants inoculated with BaC1-13. ΦPSII also decreased significantly in plants inoculated with BaC1-21, but increased significantly with BaC1-38. NPQ decreased significantly in plants inoculated with BaC1-21. The two strains able to induce systemic resistance against Xanthomonas campestris seem to work by different pathways. BaC1-13 primed enzymes related with the detoxification of reactive oxygen species (ROS). However, BaC1-38 primed pathogenesis-related proteins (PRs), and this pathway was more effective, both improved chlorophyll index confirming the priming state of the plant.

The role of peptidyl-prolyl cis/trans isomerase genes of Arabidopsis thaliana in plant defense during the course of Xanthomonas campestris infection

Experimental data obtained in this study showed the involvement of A. thaliana immunophilin genes At2g16600, At4g33060, and At5g48570 in plant defense responses to the Xanthomonas campestris invasion. We found not only that the expression levels of these genes changed upon bacterial infection, but also that the plant's resistance to the pathogen was increased if the expression levels of the immunophilin genes were elevated in the host cells.