Arabidopsis (Arabidopsis thaliana) NADPH oxidases have been reported to suppress the spread of pathogen- and salicylic acid-induced cell death. Here, we present dual roles of RBOHD (for respiratory burst oxidase homolog D) in an Arabidopsis-Alternaria pathosystem, suggesting either initiation or prevention of cell death dependent on the distance from pathogen attack. Our data demonstrate that a rbohD knockout mutant exhibits increased spread of cell death at the macroscopic level upon inoculation with the fungus Alternaria brassicicola. However, the cellular patterns of reactive oxygen species accumulation and cell death are fundamentally different in the AtrbohD mutant compared with the wild type. Functional RBOHD causes marked extracellular hydrogen peroxide accumulation as well as cell death in distinct, single cells of A. brassicicola-infected wild-type plants. This single cell response is missing in the AtrbohD mutant, where infection triggers spreading-type necrosis preceded by less distinct chloroplastic hydrogen peroxide accumulation in large clusters of cells. While the salicylic acid analog benzothiadiazole induces the action of RBOHD and the development of cell death in infected tissues, the ethylene inhibitor aminoethoxyvinylglycine inhibits cell death, indicating that both salicylic acid and ethylene positively regulate RBOHD and cell death. Moreover, A. brassicicola-infected AtrbohD plants hyperaccumulate ethylene and free salicylic acid compared with the wild type, suggesting negative feedback regulation of salicylic acid and ethylene by RBOHD. We propose that functional RBOHD triggers death in cells that are damaged by fungal infection but simultaneously inhibits death in neighboring cells through the suppression of free salicylic acid and ethylene levels.

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Leaves of powdery mildew-susceptible barley (Hordeum vulgare cv. Ingrid) and related nearisogenic lines bearing various resistance genes (Mla12, Mlg or mlo5) were inoculated with Blumeria graminis f. sp. hordei race A6. Fungal attack induced several-fold increases in ethylene emission and electrolyte leakage in leaves of susceptible Ingrid beginning 3 days after inoculation. Activities of peroxidase, superoxide dismutase, glutathione S-transferase, ascorbate peroxidase and glutathione reductase enzymes were induced markedly in susceptible leaves 5-7 days after inoculation. Similar, but less pronounced pathogen-induced changes were detected in inoculated leaves of Mla-type resistant plants that show hypersensitive cell death upon inoculation, and, to an even lesser extent, in the Mlg and mlo lines, where no visible symptoms accompanied the incompatible interaction. Glutathione content increased only in susceptible barley 7 days after inoculation. Catalase activity, total ascorbate content and redox state were not influenced by inoculation in any of the genotypes. The activity of dehydroascorbate reductase was significantly reduced 3-5 days after inoculation in the susceptible parental plants and after 5 days in Mla and Mlg lines, while it was stable in the mlo barley. Slightly elevated levels of H 2 O 2 were observed in the inoculated resistant plants. In contrast, H 2 O 2 content decreased in the susceptible line 7 days after pathogen attack. These data indicate that high levels of antioxidants are involved in the compatible interaction of susceptible barley and powdery mildew by protecting the pathogen from oxidative damage.

Membrane damage caused by the non-specific fungal toxin fusaric acid was less on pretreated than on control leaves when tobacco leaves were pretreated with anti-senescence plant hormones, such as kinetin, benzyladenine or the anti-ozonant N-[2-(2-oxo-1-imidazolidinyl)]ethyl-N'-phenylurea. Similarly, the necrosis caused by mercuric chloride was reduced by the above anti-senescence agents. In addition, in in vitro tests, leaves from selected paraquat-tolerant tobacco plants were less sensitive to Alternaria alternata (Fr) Keissler infection than those of the control paraquat-sensitive tobacco leaves. Paraquat-tolerant Conyza canadensis (L) Cronq weeds naturally selected in vineyards in Hungary showed similar inhibition of senescence to paraquat-tolerant tobacco, expressed as more green leaves and slower development. In accordance with this, the paraquat-tolerant Conyza leaves remained almost symptomless, while paraquat-sensitive plants showed severe symptoms after infection with Botrytis cinerea Pers. Oxidative burst (accumulation of hydrogen peroxide) was attenuated in TMV-infected leaves of Xanthi-nc tobacco as a result of treatment with salicylic acid or in leaves where systemic acquired resistance (SAR) had been induced by a previous TMV infection. Accordingly, superoxide dismutase (SOD) activity was higher in Xanthi tobacco leaves with SAR than without SAR. However, in NahG tobacco, in which SAR cannot develop, there was no augmentation of SOD activity. All the above data support the significance of delayed senescence and antioxidants in the resistance of plants to biotic and abiotic necrosis-inducing agents.

3Twelve bacteriophage isolates of Erwinia amylovora, the causal agent of fire blight, were isolated from blighted 4 apple, pear and quince trees from different sites in Hungary. According to morphological characteristics they 5 were assigned to the order Caudovirales, two isolates belonging to the Podoviridae and ten to the Myoviridae 6 families. Examining plaque morphology, host range and molecular characterization by PCR established that 7 these phages are not identical neither to the three North American strains used as references nor the earlier 8 isolated Hungarian Siphoviridae strains. Studying the efficacy of selected phages in apple blossoms and green 9 pear fruit slices it was found that a combination of three phage isolates (ΦEaH2A, ΦEaH5K and ΦEaH7B)

Summary Cell wall peroxidases and plasma membrane‐localized NADPH oxidases are considered to be the main sources of the apoplastic oxidative burst in plants attacked by microbial pathogens. In spite of this established doctrine, approaches attempting a comparative, side‐by‐side analysis of the functions of extracellular reactive oxygen species (ROS) generated by the two enzymatic sources are scarce. Previously, we have reported the role of Arabidopsis NADPH oxidase RBOHD (respiratory burst oxidase homologue D) in plants challenged with the necrotrophic fungus Alternaria brassicicola . Here, we present results on the activity of apoplastic class III peroxidases PRX33 ( At3g49110 ) and PRX34 ( At3g49120 ) investigated in the same Arabidopsis – Alternaria pathosystem. ROS generated by Arabidopsis peroxidases PRX33 and PRX34 increase the necrotic symptoms and colonization success of A. brassicicola . In addition, the knockdown of PRX33 and PRX34 transcript levels leads to a reduced number of host cells showing an extracellular burst of ROS after inoculation with A. brassicicola . Our results also reveal an age‐dependent transcript distribution of ROS‐producing peroxidase and NADPH oxidase enzymes, and some potential new components of the RBOHD, PRX33 and PRX34 signalling networks.

In this study transcriptomic alterations of bacterially induced pattern triggered immunity (PTI) were compared with other types of tobacco–Pseudomonas interactions. In addition, using pharmacological agents we blocked some signal transduction pathways (Ca2+ influx, kinases, phospholipases, proteasomic protein degradation) to find out how they contribute to gene expression during PTI. PTI is the first defense response of plant cells to microbes, elicited by their widely conserved molecular patterns. Tobacco is an important model of Solanaceae to study resistance responses, including defense mechanisms against bacteria. In spite of these facts the transcription regulation of tobacco genes during different types of plant bacterial interactions is not well-described. In this paper we compared the tobacco transcriptomic alterations in microarray experiments induced by (i) PTI inducer Pseudomonas syringae pv. syringae type III secretion mutant (hrcC) at earlier (6 h post inoculation) and later (48 hpi) stages of defense, (ii) wild type P. syringae (6 hpi) that causes effector triggered immunity (ETI) and cell death (HR), and (iii) disease-causing P. syringae pv. tabaci (6 hpi). Among the different treatments the highest overlap was between the PTI and ETI at 6 hpi, however, there were groups of genes with specifically altered activity for either type of defenses. Instead of quantitative effects of the virulent P. tabaci on PTI-related genes it influenced transcription qualitatively and blocked the expression changes of a special set of genes including ones involved in signal transduction and transcription regulation. P. tabaci specifically activated or repressed other groups of genes seemingly not related to either PTI or ETI. Kinase and phospholipase A inhibitors had highest impacts on the PTI response and effects of these signal inhibitors on transcription greatly overlapped. Remarkable interactions of phospholipase C-related pathways with the proteasomal system were also observable. Genes specifically affected by virulent P. tabaci belonged to various previously identified signaling routes, suggesting that compatible pathogens may modulate diverse signaling pathways of PTI to overcome plant defense.