Phenolic compounds as defence response of pepper fruits to Colletotrichum coccodes

Qualitative and quantitative changes of individual and total phenolics induced by Colletotrichum coccodes fungal infection have been studied in two susceptible sweet pepper cultivars ‘Soroksari’ and ‘Bagoly’, and the role of soluble phenolic compounds in plant's defence mechanism has been evaluated. Three distinct parts were analysed on pepper fruit: healthy tissue, anthracnose lesion, and bordering tissue, and individual phenolic compounds have been identified with the use of HPLC-MS system. In pepper fruit pericarp 21 phenolic compounds have been determined; the prevalent apigenin, quercetin and luteolin glycosides, chlorogenic acid and one chrysoeriol glucoside. C. coccodes infection increased the accumulation of chlorogenic acid, chrysoeriol glucoside, quercetin and luteolin glycosides in infected bordering tissue of both analysed pepper cultivars compared to healthy pepper tissue or symptomatic spot. Total apigenin derivatives did not show a significant increase in bordering tissue compared to the healthy pepper fruit in contrast to other groups of phenolics. This suggests a lesser role of apigenin glycosides in pepper plant defence against the Colletotrichum fungus. Intense phenolic synthesis was characteristic for the bordering zone between the healthy and infected plant tissue resulting in higher total phenolic content which might hinder the fungus to spread from the infected cells into the healthy tissue.     

Production of Indole-3-acetic Acid and Tryptophol by Pythium ultimum and Pythium Group F: Possible Role in Pathogenesis

Pythium group F is a minor pathogen which induces symptomless infections that occur frequently and results in yield losses in tomato soilless cultures. To elucidate the mode of action of this microorganism, the influence of culture filtrates of Pythium group F on tomato growth was investigated and compared to that of the pathogen Pythium ultimum. Depending on metabolite production by the fungus, marked differences were observed in plant response. Pythium group F crude culture filtrates or low molecular weight fractions (< 500) caused swelling behind the root tip and reduced root growth; the cohesion and adherence of cells within the cortical area were also affected. These symptoms were similar to those observed on plants treated with indole-3-acetic acid. By contrast, P. ultimum filtrates caused a marked distortion of cell shape accompanied with folding of host cell walls, particularly in the cortical area. These symptoms were characteristic of the activity of toxic compound(s) on host cells. Chemical analysis of the filtrates demonstrated that indole-3-acetic acid and tryptophol were produced by Pythium group F and P. ultimum. However, Pythium group F took up and metabolized more indole-3-acetic acid precursors, especially tryptophan, a key amino acid in the pathways leading to indole-3-acetic acid synthesis. The fact that Pythium group F and P. ultimum transformed tryptamine and indole-3-acetaldehyde into indole-3-acetic acid and tryptophol confirms the existence of a tryptamine pathway within both fungi. These results support the hypothesis that auxins facilitate Pythium group F infections. On the other hand, toxin(s) and hydrolytic enzymes are likely involved in P. ultimum pathogenesis.     

Correlation between lipid peroxidation and phenolics content in leaves and roots of sugar beet infected with Rhizoctonia solani

The correlation between intensity of lipid peroxidation and changes in antioxidant capacity of sugar beet plants (cv. ‘Drena’) infected with Rhizoctonia solani Kühn isolate (AG 2-2 IIIB group) was studied. Successful inoculation was confirmed by the presence of infection cushions in a cross section of leaf petioles. On the 7^th day of the experiment, phenylalanine ammonia-lyase (PAL; EC. 4.3.1.5) activity was in negative correlation with intensified lipid peroxidation process in leaves of sugar beet plants (r= –0 .99). Also, in leaves and roots of inoculated sugar beet plants, total flavonoids content (35% and 20%, respectively) and 1,1-diphenyl-2-picrylhydrazyl (DPPH)-scavenging activity (80% and 55%, respectively) were significantly reduced. Necrotic processes resulting from R. solani infection of sugar beet plants was followed by induction of plant phenolics metabolism; however, antioxidant capacity of these plants was reduced.     

Defence responses in flag leaves and spikes of common wheat Triticum aestivum cultivars with contrasting levels of basal resistance to blast caused by Pyricularia oryzae

Wheat blast, caused by Pyricularia oryzae, can cause large yield losses in crops. This study aimed to investigate defence responses in flag leaves and spikes of wheat cultivars BR-18 (moderately resistant) and BRS-Guamirim (susceptible), which differ in their levels of basal resistance. In contrast to cultivar BRS-Guamirim, infected plants of cultivar BR-18 showed more pronounced increases in activities of β-1,3-glucanase and chitinase as well as higher concentrations of lignin-thioglycolic acid derivatives in the flag leaves and total soluble phenolics in the spikes. Polyphenoloxidase activity increased in both flag leaves and spikes in response to fungal infection, regardless of cultivar. Phenylalanine ammonia-lyase (PAL) activity increased in infected flag leaves of both cultivars, especially in BR-18. PAL activity was lower in spikes of infected compared to noninfected plants of both cultivars, although to a lesser extent in BR-18. Compared to BRS-Guamirim, the antioxidative system in both flag leaves and spikes of BR-18 was more efficient in removing reactive oxygen species, reducing cellular damage caused by fungal infection. The lower catalase and peroxidase activities, associated with high superoxide dismutase activity, in flag leaves and spikes of infected BR-18 culminated in a high hydrogen peroxide concentration. The increase in ascorbate peroxidase activity was higher in both flag leaves and spikes of infected plants of BR-18 than in infected BRS-Guamirim. It was concluded that wheat resistance to blast depended on the basal level of resistance of the cultivar, which was mainly associated with the activities of defence enzymes and a more effective antioxidative system.     

Powdery mildew of Arabidopsis thaliana: a pathosystem for exploring the role of silicon in plant–microbe interactions

Silicon (Si) has been used in agriculture to protect plants against disease for hundreds of years and its prophylactic effects in monocots and dicots are well documented. The mechanisms by which Si exerts its protective effects in planta, however, are uncertain and presently the subject of debate. In this study, we sought to determine if Arabidopsis thaliana could be used to clarify the role of Si in plant–pathogen interactions. Accordingly, X-ray microanalysis mapping, light microscopy, scanning and transmission electron microscope techniques were used to examine the leaves of Si− fed A. thaliana plants inoculated with the powdery mildew fungus, Erysiphe cichoracearum. The results of this study demonstrate for the first time, that A. thaliana is a species that absorbs Si and that the incidence of powdery mildew disease for Si− fed plants is significantly lower compared to control plants. In particular, treatment with Si appeared to induce the production of an electron-dense, fungitoxic substance that accumulated within and around the collapsed fungal haustoria of infected epidermal cells within the leaves of disease-resistant plants. These results with Arabidopsis corroborate recent observations in other non-related species and support the emerging theory that the mechanisms by which Si imparts resistance to plants are complex and are not entirely explained by the traditionally proposed role of Si as a reinforcer of mechanical resistance. Collectively, the findings of the present study have established the Arabidopsis thaliana-Erysiphe cichoracearum pathosystem as a valid model to investigate the role of Si in plant–microbe interactions.     

Histochemical and Cytochemical Investigations of Phenols in Roots of Banana Infected by the Burrowing Nematode Radopholus similis

ABSTRACT The burrowing nematode Radopholus similis is one of the most damaging pathogens on banana plantations. The role of phenolics in plant defense responses to the nematode was histochemically and ultrastructurally investigated in susceptible and partially resistant cultivars. Histochemical observations of healthy roots revealed that high levels of lignin, flavonoids, dopamine, cafeic esters, and ferulic acids were associated with a very low rate of nematode root penetration in the resistant cultivar. The presence of lignified and suberized layers in endodermal cells contributed to limit invasion of the vascular bundle by the pathogen. After infection, flavonoids were seen to accumulate early in walls of cells close to the nematode-migrating channel in both cultivars and in all tissues of the infected resistant roots including the vascular tissues. The labeling pattern obtained with the gold-complexed laccase and with anti-pectin monoclonal antibodies showed that phenolics were distributed in a loosened pectin-rich material surrounding the nematode. This study provides indications that constitutive phenolics in banana roots are associated with the limitation of host penetration and colonization by R. similis. Accumulation of flavonoids in response to infection was detected in the vascular tissues of susceptible plants and in all root tissues in the partially resistant plants.     

Artificial infection of Vaccinium vitis-idaea L. and defence responses to Exobasidium species

An inoculation method for Exobasidium splendidum and Exobasidium vaccinii was developed on the dwarf shrub Vaccinium vitis-idaea. Using inoculated ramets, we investigated whether there are differences between V. vitis-idaea populations in the susceptibility to Exobasidium infections and whether the defence reaction of V. vitis-idaea is visible at a molecular level. Sixteen V. vitis-idaea clones from four populations were propagated in tissue cultures and the ramets were inoculated with E. splendidum or E. vaccinii fungi. The expression of three flavonoid biosynthetic genes (chalcone synthase, dihydroflavonol 4-reductase and anthocyanidin synthase) and the accumulation of flavonoids and hydroxycinnamic acids were determined in response to E. splendidum infection. A pathogenesis-related (PR 4) gene was isolated and its expression was studied in host ramet leaves. To our knowledge, this was the first successful artificial infection reported with E. splendidum. Disease frequencies of the inoculated ramets were between 32% and 47% for E. splendidum and 33% for E. vaccinii, but below 10% in uninoculated control ramets. There were no differences in disease frequencies between V. vitis-idaea populations. Both symptomatic leaves and healthy leaves of diseased ramets showed activation of flavonoid biosynthesis at the gene level, whereas expression of PR 4 was observed only in symptomatic leaves. The increase of flavonoid biosynthesis in healthy leaves of diseased ramets may represent a general response to stress or a role in defence against the pathogen E. splendidium. Ability of V. vitis-idaea to defend chemically against Exobasidium fungi and the heterogeneity of genotypes, age, size, and growth rates in host plant populations might be reasons for the low infection incidence of Exobasidia in nature.     

Moringa Oleifera—Establishment and Multiplication of Different Ecotypes In Vitro

To obtain healthy plant material from Moringa oleifera regardless of season, weather, and degree of infestation, a procedure to establish an in vitro culture of M. oleifera from nodes using 0.2 % mercury chloride was developed. It was not possible to create an in vitro culture of M. oleifera from seeds. Nodes were cultivated on MS medium with different concentrations of benzylaminopurine (BAP) and agar contents to find the best conditions for rapid growth and optimum multiplication. The highest multiplication ability of the different plant parts, especially the base parts of M. oleifera in vitro plants, was observed after 3 weeks of cultivation on MS medium with 0.5 mg l^?1 BAP. Callus formation increases with increased BAP concentration (0, 0.5, 0.75, 1 mg l^?1 BAP). Furthermore, the use of two phytohormones, indole-3-acetic acid and thidiazuron—led to very strong callus formation of adaxial and abaxial orientated leaves on MS medium. This formation was only observed for material that was light induced for 24 h prior to cultivation under dark conditions. Analysis of the glucosinolate content of M. oleifera leaves revealed a different glucosinolate profile of plants cultivated in vitro and in soil beds in the greenhouse. Whereas in greenhouse leaves rhamnopyranosyloxy-benzyl glucosinolates were abundant, the precursor benzyl glucosinolate was found in in vitro cultures.     

Biochemical and physiological responses to long‐term Citrus tristeza virus infection in Mexican lime plants

Reactions that occur when a plant is subjected to Citrus tristeza virus (CTV) infection often result in triggering of numerous defence mechanisms to fight the infection. The reactions vary according to virus strain, host genotype, time of exposure to the infection and environmental conditions. To date, no study has examined in detail the consequences of 10‐year exposure to CTV infection on the biochemical and physiological status of susceptible Mexican lime plants (Citrus aurantifolia). To understand the reaction of such plants, changes in nutrient status, total proteins, enzyme activity involved in scavenging of reactive oxygen species, photosynthetic and transpiration rates, chlorophyll content, membrane permeability and water content were analysed in plants infected with different CTV isolates and in healthy plants. The activity of superoxide dismutase and polyphenol oxidase significantly decreased in the infected leaves, and membrane permeability was lower in the infected plants. Macro‐ and micronutrient elements were significantly changed: concentrations of leaf nitrogen, zinc, magnesium and iron were elevated but potassium concentration depressed in comparison to noninfected control leaves. Levels of other analysed nutrient elements, enzymes, photosynthesis and stomatal conductance, chlorophyll content and relative water content were unchanged. Clear physiological changes were found among infected and noninfected control plants but none between plants infected with different CTV isolates. The data suggest that some of the defence mechanisms investigated here were suppressed due to the continuous and long‐term pressure of biotic stress.     

Engineering flax with increased flavonoid content and thus Fusarium resistance

Flavonoids are a group of secondary plant metabolites important for plant growth and development, and thus the regulation of their biosynthesis is of special interest. We used a transgenic approach for flavonoid content manipulation. The multigene construct contained the cDNAs for chalcone synthase (CHS), and chalcone isomerase (CHI) and dihydroflavonol reductase (DFR) were prepared. Following flax plants transformation, the levels of the products of the enzyme overproduction were assessed in leaves and seeds. The simultaneous expression of genes resulted in a significant increase in the levels of flavanones, flavones, flavonols and anthocyanins, suggesting those three overproducing enzymes efficiently control the flavonoid route of the phenylpropanoid pathway.The increase in the flavonoid content in the transgenic flax plants might be the reason for observed, enhanced antioxidant capacity of those plants. The increased antioxidative properties of transgenic plants lead to improved resistance to Fusarium, the main pathogen of flax.The changes in phenylpropanoids accumulation in transgenic plants affect cell wall carbohydrate content. Immunochemical studies revealed significant increase in carbohydrates, constituents of pectin and hemicellulose. Since pectins contribute to flax stem retting, the compounds increase might affect fibre production. An increase in pectin and hemicellulose content leads to enhanced disease resistance of those plants.     

Reduction of clubroot (Plasmodiophora brassicae) formation in Arabidopsis thaliana after treatment with prohexadione‐calcium,an inhibitor of oxoglutaric acid‐dependent dioxygenases

Recently, flavonoids were shown to modulate the outcome of clubroot development in Arabidopsis thaliana after infection with the obligate biotrophic pathogen Plasmodiophora brassicae. Therefore, the development of clubroot disease was investigated in Arabidopsis after treatment with prohexadione‐calcium (ProCa), an inhibitor of ascorbic acid/2‐oxoglutaric acid‐dependent dioxygenases such as flavanone‐3‐hydroxylase. The treatment resulted in a reduction of the flavonols quercetin and kaempferol in clubroots, whereas the precursor naringenin highly accumulated. The root system of ProCa‐treated plants was better developed although galls were still visible. Thus, ProCa treatment resulted in reduced gall size. Flavonoids are thought to inhibit polar auxin transport by modulating auxin efflux carriers. It was investigated whether the auxin response might change as a consequence of the accumulation of naringenin in ProCa‐treated plants. In the areas of gall development an auxin response was indicated by the auxin‐responsive promoter DR5 coupled to the reporter β‐glucuronidase (GUS), whereas very little staining was found in healthy root parts. No differences in GUS activity were found between P. brassicae‐infected and ProCa‐treated plants, and plants only infected with P. brassicae, indicating that the effect of ProCa treatment on clubroot reduction is not via changes in auxin responses. As ProCa is also an inhibitor of late steps in gibberellin biosynthesis, a specific gibberellin biosynthesis inhibitor, chlormequatchloride (CCC), was tested on club development. However, CCC did not reduce disease symptoms, indicating that the observed reduced gall development was not because of gibberellin biosynthesis inhibition by ProCa.     

Expression patterns of defence genes and antioxidant defence responses in a rice variety that is resistant to leaf blast but susceptible to neck blast

The rice stage resistance variety Jiajing3768 is resistant to leaf blast but susceptible to neck blast. This variety was used to analyze the expression patterns of defence genes and antioxidant defence responses in the leaves at the seedling stage (LSS) and necks at the preliminary heading stage (NPHS), after inoculation with Magnaporthe oryzae. We found that defence genes PR1b (pathogenesis-related class 1b), PR4, PR10a, JIOsPR10 (jasmonic acid induced rice PR 10), Gns1 (1,3; 1,4-β-glucanase), Cht-1 (chitinase), and LOX (lipoxygenase) may play roles in the stage resistance in Jiajing3768; PAL (phenylalanine ammonia-lyase) and CHS (chalcone synthase) could participate in defending Jiajing3768 against neck blast but not leaf blast. The antioxidant enzymes superoxide dismutase, peroxidase, catalase, glutathione reductase, and malondialdehyde coordinately participate in the stage resistance to blast in Jiajing3768, and that oxidative damage is less in the LSS than in the NPHS.     

The influence of silicon on cytological interactions between Sphaerotheca fuliginea and Cucumis sativus

The response of epidermal cells of cucumber leaf tissue infected by Sphaerotheca fuliginea was examined by light microscopy to understand how silicon in infected host cells affects host defence mechanisms. Leaf pieces from plants treated with nutrient solutions containing 0·05, 0·50 or 2·3 mm of silicon (Si) were harvested at intervals of 24, 48, 72, 96 and 120 h after inoculation with the pathogen and examined after staining with toluidine blue or aniline blue. Si treatments significantly reduced the time to initiation of production and/or accumulation of phenolic materials in infected host epidermal cells, and increased the number of infected cells that produced and/or accumulated phenolics. The number of haustoria produced per colony of S. fuliginea was significantly reduced over time, and conidiophore development was delayed on the leaves of cucumber treated with 2·3 mm Si nutrient solution.     

Pathogenicity of Aster Yellows Phytoplasma and Spiroplasma citri on Periwinkle

ABSTRACT Using Murashige and Skoog (MS) as a basal medium, the effects of varying levels and combinations of plant growth regulators required for shoot tip and root proliferation in healthy and aster yellows phytoplasma (AYP)- and Spiroplasma citri-infected periwinkle (Catharanthus roseus) shoots were studied. Number of shoots and fresh and dry mass of healthy and AYP-infected shoots increased when benzyladenine (BA) concentrations were increased from 0.5 to 4 mg/liter. The maximum number of shoots for both healthy and AYP-infected plants was obtained when grown in MS medium supplemented with BA at 4 mg/liter and indole-3-acetic acid (IAA) at 0.5 mg/liter. S. citri-infected shoots proliferated the most when grown in MS medium containing BA at 2 mg/liter and IAA at 0.5 mg/liter. The best medium for root production in healthy periwinkle shoots contained alpha-naphthaleneacetic acid (NAA) at 0.5 mg/liter, whereas the best medium for AYP-infected shoots contained indole-3-butyric acid (IBA) at 2.5 mg/liter, both in combination with kinetin at 0.1 mg/liter. S. citri-infected shoots had the best root growth when grown in medium supplemented with IBA at 5.0 mg/liter and kinetin at 0.1 mg/liter. The concentration of cytokinin and auxin needed for maximum shoot proliferation differed between AYP- and S. citri-infected shoot tips, strongly indicating that the two mollicutes may cause different changes in endogenous cytokinin and auxin levels. The concentrations of NAA and IBA needed for root growth of S. citri-infected shoots were two- to fivefold higher than the concentrations needed for healthy and AYP-infected shoots, clearly demonstrating that S. citri infection caused a shortage of auxins that resulted in retardation of secondary root growth. Chlorophyll content was markedly reduced in periwinkles infected with AYP or S. citri compared with chlorophyll in healthy periwinkles. AYP caused a decrease in carotenoid in leaves 6 weeks after graft-inoculation, but carotenoid content was unchanged in S. citri-infected leaves throughout the test period. Anthocyanin content in periwinkles infected with AYP decreased significantly by 4 weeks postinoculation, whereas anthocyanin content in periwinkles infected with S. citri increased. Anthocyanin content in leaf tissues, however, was reduced as a result of AYP and S. citri infection. Pigment changes induced by AYP and S. citri, whether similar or different compared with those of healthy periwinkle shoots, provide important information for interpreting pathogenesis when linked with plant growth regulators.     

The effect of fungal infection and nitrogen fertilization on the carbohydrate composition of Rumex obtusifolius leaves

The effect of infection by the rust fungus Uromyces rumicis and fertilization by different concentrations of nitrate or ammonium solutions on the concentrations of alcohol-soluble carbohydrates, fructans and starch in Rumex obtusifolius leaves was investigated. In leaves from healthy plants there was an increase in concentration of alcohol-soluble carbohydrates and fructans as the concentration of nitrate given was decreased, with the exception of nitrogen-stressed plants (those fed less than 10 mM l^-1 nitrate) which had a lower concentration of alcohol-soluble carbohydrates than plants fed 10 mM l^-1 nitrate. There was an increase in fructan and starch concentration in leaves as the concentration of ammonium solution given was decreased. The concentration of alcohol-soluble carbohydrates was reduced in infected leaves and increased in healthy leaves on infected plants, while the fructan concentration increased in infected leaves, compared to healthy leaves. The effects of infection were consistent over the range of nitrogen concentrations used, and thus were additive to the effects of fertilization. These results confirm the known effects of fertilization or fungal infection, singly, and indicate that, when combined, nitrogen deficiency and fungal infection may produce an additive stress on the plant.     

Biochemical response of grapevine variety ‘Chardonnay’ (Vitis vinifera L.) to infection with grapevine yellows (Bois noir)

This study was carried out on the leaves of phytoplasma susceptible grapevine variety ??Chardonnay?? (Vitis vinifera L.), and included research of the alterations in chlorophyll and carotenoid contents, contents of phenol compounds and in related enzymes activity in the phenylpropanoid pathway during the Bois noir (BN) infection. Phytoplasma-infected leaves showed reduced contents of chlorophylls and carotenoids, which promoted their susceptibility to oxidative reactions. Furthermore, modulation of flavonoid biosynthesis occurred in infected leaves, leading to an increased activity of phenylalanine ammonia lyase, chalcone synthase/chalcone isomerase, flavanone 3-hydroxylase and polyphenoloxidase, but to a decreased peroxidase activity. Phytoplasma infection led to an increase of the contents of hydroxycinnamic acids (caftaric acid, sinapic acid glucose derivate and coutaric acid), flavanols (procyanidin B1, procyanidin dimer 3, catechin, epicatechin) and flavonols (quercetin 3-O-glucuronide, quercetin 3-O-glucoside) especially in the period up to vérasion. The study demonstrated that at certain phenological key-stages infection with phytoplasma (BN) induced different alterations in enzyme activities and in the contents of biochemical compounds from primary and secondary metabolism.     

西花蓟马取食、机械损伤和外源物质诱导对番茄植株次生物质及西花蓟马解毒酶的影响

为阐明寄主植物番茄受害虫西花蓟马Frankliniella occidentalis取食、机械损伤及外源物质茉莉酸和水杨酸甲酯诱导后次生物质含量的变化,以及害虫如何通过调整体内解毒酶活性适应植物的防御反应,采用生化分析法测定了各诱导处理下次生物质黄酮、总酚和单宁含量及西花蓟马体内解毒酶多功能氧化酶(MFO)、乙酰胆碱酯酶(AchE)、谷胱甘肽S-转移酶(GSTs)和羧酸酯酶(CarE)活性的变化。结果表明:番茄叶片在各诱导处理下黄酮含量在36 h和48 h时均显著高于对照,相应时间段茉莉酸处理与水杨酸甲酯处理的黄酮含量升高最显著,分别达32.07 mg/g和31.76 mg/g;各诱导处理后24 h和36 h时,单宁和总酚含量均有不同程度下降,但总酚含量在48 h时显著升高,其中虫害处理增加最显著,达34.51 mg/g。取食各种诱导处理番茄植株6 h时,西花蓟马体内MFO活性均升高,AchE活性均受到抑制;取食除机械损伤外的其它诱导处理叶片6 h后,西花蓟马体内GSTs活性均被抑制,CarE活性均显著上升,其中取食水杨酸甲酯处理叶片的西花蓟马体内GSTs活性下降幅度最大,而CarE活性升高最显著,分别为3 882.35 U/mg和106.33 U/mg。表明各诱导处理不仅可以使番茄次生物质含量发生变化,诱导植株产生防御反应,也会使西花蓟马改变解毒酶的活性以适应寄主植物的诱导抗性,即寄主植物和害虫通过防御与反防御相互适应。     

Priming as a Mechanism in Induced Systemic Resistance of Plants

Induced systemic resistance is a plant defence state that is associated with an enhanced ability – the so-called priming – to resist pathogen attack by stronger activation of cellular defence responses. So far, however, priming has not been widely appreciated when studying induced plant disease resistance. During the past several years, it has been demonstrated that pre-treatment of cultured parsley cells with inducers of systemic resistance, salicylic acid or a benzothiadiazole, leads to the direct activation of a set of defence-related genes and also primes the cells for stronger elicitation of another set of defence genes including those encoding phenylalanine ammonia-lyase. From these results, it was concluded that the resistance inducers have at least a dual role in plant defence-gene activation. When elucidating whether priming plays a role in induced systemic resistance of Arabidopsis, pre-treating plants with benzothiadiazole was found to augment the subsequent activation of phenylalanine ammonia-lyase genes by Pseudomonas infection, wounding and osmotic stress and also to enhance wound/osmotic stress-induced callose production. The augmentation of phenylalanine ammonia-lyase gene activation or/and callose deposition was not seen in the Arabidopsis non-expresser of pathogenesis-related genes1 mutant which is compromised in induced resistance, while it was present, without benzothiadiazole pre-treatment, in the constitutive expresser of pr genes1 and 5 mutants in which induced resistance is constitutive. Together these studies point to priming as an important cellular mechanism in induced systemic resistance of plants which requires the intact non-expresser of pathogenesis-related genes1 gene.