Development Of Disease Symptoms Research Articles

The Arabidopsis altered in stress response2 is Impaired in Resistance to Root and Leaf Necrotrophic Fungal Pathogens.

The Arabidopsis thaliana Glutathione S-transferase Phi8 (GSTF8) gene is recognised as a marker for early defence and stress responses. To identify regulators of these responses, a forward genetic screen for Arabidopsis mutants with up-regulated GSTF8 promoter activity was conducted by screening a mutagenized population containing a GSTF8 promoter fragment fused to the luciferase reporter gene (GSTF8:LUC). We previously identified several enhanced stress response (esr) mutants from this screen that conferred constitutive GSTF8:LUC activity and increased resistance to several pathogens and/or insects pests. Here we identified a further mutant constitutively expressing GSTF8:LUC and termed altered in stress response2 (asr2). Unlike the esr mutants, asr2 was more susceptible to disease symptom development induced by two necrotrophic fungal pathogens; the root pathogen Fusarium oxysporum, and the leaf pathogen Alternaria brassicicola. The asr2 allele was mapped to a 2.1 Mbp region of chromosome 2 and narrowed to four candidate loci.

Screening of a multi-virus resistant RNAi construct in cowpea through transient vacuum infiltration method.

Plant viruses are the most devastating pathogens causing substantial economic losses in many crops. Current viral disease management relies on prophylactics, roguing and insect vector control, since in most crops resistant gene pools for resistance breeding are unavailable. RNA interference, a sequence dependent gene silencing mechanism holds great potential in imparting virus resistance. In this study, the efficacy of a RNAi gene construct developed against four viruses commonly infesting tomato and chilli viz., capsicum chlorosis virus, groundnut bud necrosis virus, cucumber mosaic virus and chilli veinal mottle virus was evaluated. A 3546bp dsRNA-forming construct comprising sense-intron-antisense fragments in binary vector pBI121 (hpRNAi-MVR) was mobilized into Agrobacterium tumefaciens. Cowpea (Vigna unguiculata) was used as an indicator plant for GBNV agroinfiltration to evaluate the efficacy of hpRNAi-MVR construct in conferring GBNV resistance. The type of agroinfiltration, bacterial concentration and incubation-temperatures were optimized. Vacuum infiltration of three pulses of 20-30s at 66.66kPa were effective than syringe infiltration. Of the five Agrobacterial concentrations, OD600 0.5 was more efficient. Incubation temperature of 31 ± 1°C was favorable for development of disease symptoms than 20 ± 1°C and 26 ± 1°C. ELISA revealed a 35% decline in viral load in hpRNAi-MVR infiltrated plants compared to vector control plants. Quantitative real time PCR results have shown a viral gene silencing to the extent of 930-990 folds in hpRNAi-MVR infiltrated plants compared to vector control. This approach is simple, rapid and efficient to screen the efficacy of RNAi constructs developed for the RNAi mediated plant virus management.

Zmeny druhového spektra fuzariózy klasov spôsobené fungicídmi

Fusarium head blight (FHB) is a dangerous disease of cereals in case of yield losses and mycotoxin production. The aim of this work was to estimate the influence of used fungicides on species spectrum associated with Fusarium head blight (FHB). The study was realised in Slovakia on winter wheat by natural infection of FHB. The ears were sprayed with fungicides at beginning of flowering. After development of disease symptoms, fungicide efficacy was estimated, and Fusarium species spectrum were isolated and determined in laboratory. The fungicide efficacy against FHB achieved 60.2 – 88.3 %. The highest effectivity was estimated by using of prothioconazole + tebuconazole and prothioconazole + fluoxastrobin, followed by dimoxystrobin + cyproconazole, metconazole, propiconazole + cyproconazole, and prochloraz + tebuconazole. F. graminearum was the most frequently isolated from the ears, followed by M. nivale, F. avenaceum, F. culmorum, and F. poae. Using of prothioconazole + tebuconazole, metconazole, prothioconazole + fluoxastrobin, and tebuconazole significantly decreased incidence of F. graminearum on infected ears. The incidence of M. nivale on ears was significantly increased by using of all fungicides in comparison with untreated control. The results suggest that the ear spraying by triazole fungicides could enhance the incidence of less susceptible Fusarium species in ears, especially M. nivale. The variability in fungicide efficacy of fungicides against FHB in agricultural praxis colud be affected besides multi-pathogen origin of FHB just by selective effect of fungicides on different Fusarium species.

Small RNA analysis reveals different profiles among ‘Candidatus Phytoplasma mali’ strains in Malus x domestica cultivar Golden Delicious in vitro plants

In plant hosts phytoplasmas were shown to affect the expression of a broad range of genes and RNA regulatory molecules acting at a post-transcriptional level and leading to changes in several metabolic pathways. Effector proteins secreted by phytoplasmas play a paramount role in the metabolic changes resulting in disease symptoms development. It is shown here that in ‘Candidatus Phytoplasma mali’-infected in vitro plants of Malus x domestica cultivar Golden Delicious another class of pathogen-derived molecules represented by small RNA can be detected through specific sequencing. The profiles obtained from seven different phytoplasma strains revealed differences that could be associated to their capacity to multiply in the plant host or due to differences in the expression level of the phytoplasma transcripts.

NtPR1a regulates resistance to Ralstonia solanacearum in Nicotiana tabacum via activating the defense-related genes

Pathogenesis-related proteins (PRs) are associated with the development of systemic acquired resistance (SAR) against further infection enforced by fungi, bacteria and viruses. PR1a is the first PR-1 member that could be purified and characterized. Previous studies have reported its role in plants’ resistance system against oomycete pathogens. However, the role of PR1a in Solanaceae plants against the bacterial wilt pathogen Ralstonia solanacearum remains unclear. To assess roles of NtPR1a in tobacco responding to R. solanacearum, we performed overexpression experiments in Yunyan 87 plants (a susceptible tobacco cultivar). The results illuminated that overexpression of NtPR1a contributed to improving resistance to R. solanacearum in tobacco Yunyan 87. Specifically speaking, NtPR1a gene could be induced by exogenous hormones like salicylic acid (SA) and pathogenic bacteria R. Solanacearum. Moreover, NtPR1a-overexpressing tobacco significantly reduced multiple of R. solanacearum and inhibited the development of disease symptoms compared with wild-type plants. Importantly, overexpression of NtPR1a activated a series of defense-related genes expression, including the hypersensitive response (HR)-associated genes NtHSR201 and NtHIN1, SA-, JA- and ET-associated genes NtPR2, NtCHN50, NtPR1b, NtEFE26, and Ntacc oxidase, and detoxification-associated gene NtGST1. In summary, our results suggested that NtPR1a-enhanced tobacco resistance to R. solanacearum may be mainly dependent on activation of the defense-related genes.

Seed-born Burkholderia glumae Infects Rice Seedling and Maintains Bacterial Population during Vegetative and Reproductive Growth Stage.

Rice world production is affected due to the growing impact of diseases such as bacterial panicle blight, produced by Burkholderia glumae. The pathogen-induced symptoms include seedling rot, grain rot and leaf-sheath browning in rice plants. It is currently recognized the entrance of this pathogen to the plant, from infected seeds and from environmental sources of the microorganism. However, it is still not fully elucidated the dynamics and permanence of the pathogen in the plant, from its entry until the development of disease symptoms in seedlings or panicles. In this work it was evaluated the infection of B. glumae rice plants, starting from inoculated seeds and substrates, and its subsequent monitoring after infection. Various organs of the plant during the vegetative stage and until the beginning of the reproductive stage, were evaluated. In both inoculation models, the bacteria was maintained in the plant as an endophyte between 1 × 101 and 1 × 105 cfu of B. glumae.g−1 of plant throughout the vegetative stage. An increase of bacterial population towards initiation of the panicle was observed, and in the maturity of the grain, an endophyte population was identified in the flag leaf at 1 × 106 cfu of B. glumae.g−1 fresh weight of rice plant, conducting towards the symptoms of bacterial panicle blight. The results found, suggest that B. glumae in rice plants developed from infected seeds or from the substrate, can colonize seedlings, establishing and maintaining a bacterial population over time, using rice plants as habitat to survive endophyticly until formation of bacterial panicle blight symptoms.

Evaluation of Malus gene bank resources with German strains of Marssonina coronaria using a greenhouse-based screening method

Marssonina coronaria is considered a threat to organic apple production in Central Europe. Since the application of fungicides is limited in organic production, breeding for resistance seems to be a promising strategy to manage the disease in the future. In this study, an artificial inoculation method similar to procedures used in apple scab greenhouse screenings was developed using German strains of the pathogen M. coronaria for evaluating 110 Malus domestica cultivars. The strains were morphologically and molecularly characterized and confirmed as M. coronaria. Symptom development was significantly influenced by incubation method, conidia concentration and point of inoculation, but not by leaf position and different water types used for inoculation. The success of inoculation and spread of the fungus on infected leaves were confirmed by conventional PCR. Moreover, there was a difference in development of disease symptoms between inoculations with conidia from in vitro grown strains (in vitro inoculum) and conidia from diseased leaves from the orchard (field inoculum) in a time dependent manner. These differences which were first found with the susceptible cultivar ‘Topaz’ after artificial inoculation with in vitro and field inoculum were confirmed on 20 more apple cultivars inoculated with different in vitro grown strains. In summary, all tested cultivars, including 21 which are scab-resistant, developed symptoms of this disease. Results from three years of investigation indicate a decrease in virulence of M. coronaria strains, when cultivated on artificial culture (growth)-media. Hence field inoculum is recommended for artificial greenhouse screenings for the evaluation of disease resistance in Malus genetic resources.

Molecular Profiling of Pierce's Disease Outlines the Response Circuitry of Vitis vinifera to Xylella fastidiosa Infection.

Pierce’s disease is a major threat to grapevines caused by the bacterium Xylella fastidiosa. Although devoid of a type 3 secretion system commonly employed by bacterial pathogens to deliver effectors inside host cells, this pathogen is able to influence host parenchymal cells from the xylem lumen by secreting a battery of hydrolytic enzymes. Defining the cellular and biochemical changes induced during disease can foster the development of novel therapeutic strategies aimed at reducing the pathogen fitness and increasing plant health. To this end, we investigated the transcriptional, proteomic, and metabolomic responses of diseased Vitis vinifera compared to healthy plants. We found that several antioxidant strategies were induced, including the accumulation of gamma-aminobutyric acid (GABA) and polyamine metabolism, as well as iron and copper chelation, but these were insufficient to protect the plant from chronic oxidative stress and disease symptom development. Notable upregulation of phytoalexins, pathogenesis-related proteins, and various aromatic acid metabolites was part of the host responses observed. Moreover, upregulation of various cell wall modification enzymes followed the proliferation of the pathogen within xylem vessels, consistent with the intensive thickening of vessels’ secondary walls observed by magnetic resonance imaging. By interpreting the molecular profile changes taking place in symptomatic tissues, we report a set of molecular markers that can be further explored to aid in disease detection, breeding for resistance, and developing therapeutics.

Environmental and inoculum effects on epidemiology of bacterial spot disease of stone fruits and development of a disease forecasting system

Bacterial spot disease of stone fruits, caused by Xanthomonas arboricola pv. pruni, is of high economic importance in the major stone-fruit-producing areas worldwide. A better understanding of disease epidemiology can be valuable in developing disease management strategies. The effects of weather variables (temperature and wet/dry period) on epiphytic growth of X. arboricola pv. pruni on Prunus leaves were analyzed, and the relationship between inoculum density and temperature on disease development was determined and modeled. The information generated in this study, performed under controlled environmental conditions, will be useful to develop a forecasting system for X. arboricola pv. pruni. Optimal temperature for growth of epiphytic populations ranged from 20 to 30 °C under leaf wetness. In contrast, multiplication of epiphytic populations was not only interrupted under low relative humidity (RH) (< 40%) at 25 °C, but also resulted in cell inactivation, with only 0.001% initial cells recovered after 72 h incubation. A significant effect of inoculum density on disease severity was observed and 106 CFU/ml was determined as the minimal infective dose for X. arboricola pv. pruni on Prunus. Infections occurred at temperatures from 15 to 35 °C, but incubation at 25 and 30 °C gave the shortest incubation periods (7.7 and 5.9 days respectively). A model for predicting disease symptom development was generated and successfully evaluated, based on the relationship between disease severity and the accumulated heat expressed in cumulative degree day (CDD). Incubation periods of 150, 175 and 280 CDD were required for 5, 10 and 50% of disease severity, respectively.

Потенциальные возможности применения нутрицевтических продуктов с целью пролонгирования ремиссии у пациентов с синдромом раздраженного кишечника

Aim of review. To summarize the modern data on pathogenesis of symptoms irritable bowel syndrome (IBS) symptoms, to present nutraceutical product «Standart zdorovya: GASTRO» as an agent for IBS remission maintenance. Summary. Recent data indicate essential changes of intestinal microbiota in IBS in comparison to healthy population. These changes may be associated to development of inflammatory processes of intestinal wall. Long-term inflammation may result in nociceptor hypersensitivity i.e. peripheral sensitization that in turn, lead to the central sensitization and indirectly - to gastrointestinal tract (GIT) motility disorders. Besides, change of intestinal microbiota may impact the development of emotional disorders (e.g.: impaired capacity of affected intestinal microbiota to produce neurotransmitters). Therefore, according to the results of available studies, intestinal microbiota changes in IBS is a key factor for the cascade of events including intestinal wall inflammation, changes of gastrointestinal motility, decreased threshold of gastrointestinal receptor sensitivity, as well as emotional disorders resulting in development of disease symptoms. Medical preparation for IBS remission induction and remission maintenance should possess probiotic/prebiotic and anti-inflammatory properties, reduce visceral sensitivity, normalize intestinal motility and modulate patients’ emotional sphere. Due to capacity of «Standart zdorovya: GASTRO» components to deliver above-listed effects it can be promising for administration in patients with IBS.

Two ancestral genes shaped the Xanthomonas campestris TAL effector gene repertoire.

Xanthomonas transcription activator-like effectors (TALEs) are injected inside plant cells to promote host susceptibility by enhancing transcription of host susceptibility genes. TALE-encoding (tal) genes were thought to be absent from Brassicaceae-infecting Xanthomonas campestris (Xc) genomes based on four reference genomic sequences. We discovered tal genes in 26 of 49 Xc strains isolated worldwide and used a combination of single molecule real time (SMRT) and tal amplicon sequencing to yield a near-complete description of the TALEs found in Xc (Xc TALome). The 53 sequenced tal genes encode 21 distinct DNA binding domains that sort into seven major DNA binding specificities. In silico analysis of the Brassica rapa promoterome identified a repertoire of predicted TALE targets, five of which were experimentally validated using quantitative reverse transcription polymerase chain reaction. The Xc TALome shows multiple signs of DNA rearrangements that probably drove its evolution from two ancestral tal genes. We discovered that Tal12a and Tal15a of Xcc strain Xca5 contribute together in the development of disease symptoms on susceptible B.oleracea var. botrytis cv Clovis. This large and polymorphic repertoire of TALEs opens novel perspectives for elucidating TALE-mediated susceptibility of Brassicaceae to black rot disease and for understanding the molecular processes underlying TALE evolution.

Advances on Fungal Phytotoxins and Their Role in Grapevine Trunk Diseases

Grapevines are produced worldwide with important impact on local economies. Several biotic stresses induce serious diseases of grapevine, which severely affect the quantity and quality of production. One of the most important problems of vineyards worldwide is the high incidence of grapevine trunk diseases (GTD) induced by fungi belonging to several genera. Environmentally friendly methods for GTD control are being studied. This perspective offers an advanced overview on the fungal phytotoxins involved in GTD and their eventual role in the development of disease symptoms.

First Report of Two Colletotrichum Species Associated with Bitter Rot on Apple Fruit in Korea – C. fructicola and C. siamense

Bitter rot caused by the fungal genus Colletotrichum is a well-known, common disease of apple and causes significant yield loss. In 2013, six fungal strains were isolated from Fuji apple fruits exhibiting symptoms of bitter rot from Andong, Korea. These strains were identified as Colletotrichum fructicola and C. siamense based on morphological characteristics and multilocus sequence analysis of the internal transcribed spacer rDNA, actin, calmodulin, chitin synthase, and glyceraldehyde-3-phosphate dehydrogenase Pathogenicity tests confirmed the involvement of C. fructicola and C. siamense in the development of disease symptoms on apple fruits. This is the first report of C. fructicola and C. siamense causing bitter rot on apple fruit in Korea.

Protection from chronic stress- and depressive symptom-induced vascular endothelial dysfunction in female rats is abolished by preexisting metabolic disease.

While it is known that chronic stress and clinical depression are powerful predictors of poor cardiovascular outcomes, recent clinical evidence has identified correlations between the development of metabolic disease and depressive symptoms, creating a combined condition of severely elevated cardiovascular disease risk. In this study, we used the obese Zucker rat (OZRs) and the unpredictable chronic mild stress (UCMS) model to determine the impact of preexisting metabolic disease on the relationship between chronic stress/depressive symptoms and vascular function. Additionally, we determined the impact of metabolic syndrome on sex-based protection from chronic stress/depressive effects on vascular function in female lean Zucker rats (LZRs). In general, vasodilator reactivity was attenuated under control conditions in OZRs compared with LZRs. Although still impaired, conduit arterial and resistance arteriolar dilator reactivity under control conditions in female OZRs was superior to that in male or ovariectomized (OVX) female OZRs, largely because of better maintenance of vascular nitric oxide and prostacyclin levels. However, imposition of metabolic syndrome in combination with UCMS in OZRs further impaired dilator reactivity in both vessel subtypes to a similarly severe extent and abolished any protective effect in female rats compared with male or OVX female rats. The loss of vascular protection in female OZRs with UCMS was reflected in vasodilator metabolite levels, which closely matched those in male and OVX female OZRs subjected to UCMS. These results suggest that presentation of metabolic disease in combination with depressive symptoms can overwhelm the vasoprotection identified in female rats and, thereby, may reflect a severe impairment to normal endothelial function. NEW & NOTEWORTHY This study addresses the protection from chronic stress- and depression-induced vascular dysfunction identified in female compared with male or ovariectomized female rats. We determined the impact of preexisting metabolic disease, a frequent comorbidity of clinical depression in humans, on that vascular protection. With preexisting metabolic syndrome, female rats lost all protection from chronic stress/depressive symptoms and became phenotypically similar to male and ovariectomized female rats, with comparably poor vasoactive dilator metabolite profiles.

Characterization of p-Coumaric acid-induced soluble and cell wall-bound phenolic metabolites in relation to disease resistance to Xanthomonas campestris pv. campestris in Chinese cabbage

To characterize the p-coumaric acid (pCA)-induced phenolic metabolites in relation to the disease resistance against Xanthomonas campestris pv. campestris (Xcc.), the responses of soluble- and cell wall-bound flavonoid and hydroxycinnamic acids compounds to the pretreatment of pCA or the inhibitor of the 4-coumarate–CoA ligase, 3,4-(methylenedioxy) cinnamic acid (MDCA), following Xcc inoculation were assessed, and the resulting data were interpreted with regard to susceptibility to Xcc in Chinese cabbage (Brassica rapa var. pekinensis). At 12 days post-inoculation (DPI) with Xcc, disease symptom development could be distinguished by necrotic lesions, and characterized by an enhanced lipid peroxidation. Overall, pCA acts as a positive stimulus for an accumulation of hydroxycinnamic acids and flavonoids, while MDCA acts as a negative regulator. Pretreatment with pCA resulted in an accumulation of specific hydroxycinnamic acids, pCA, ferulic acid (FA), and sinapic acid (SiA) in both soluble and cell wall-bound forms in Xcc-inoculated leaves, while MDCA pretreatment decreased accumulation in a dose-dependent manner. Two flavonoid compounds, epigallocatechin (EGC) and epigallocatechin gallate (EGCG), showed a similar response to pCA and MDCA pretreatments. These results indicate that a lower disease symptom development in pCA-pretreated leaves was associated with a higher accumulation of hydroxycinnamic acids and flavonoids, and vice-versa in MDCA- and non-pretreated (control) leaves.

A canker disease of apple caused by Cytospora parasitica recorded in China

SummaryA stem canker disease of apple that differs from previously reported canker diseases has been recorded in Xinjiang Uygur Autonomous Region in China. Eight Cytospora strains obtained from diseased apple trees and inoculated onto healthy apple tree branches resulted in the development of the same disease symptoms. This pathogen formed conceptacles, which were multilocular pycnidia with common walls and a single ostiole. Comprehensive analysis of three DNA sequence regions (internal transcribed spacer, large ribosomal subunit and transcription elongation factor) and morphological and cultural characteristics revealed that these strains were most similar to the fungus Cytospora parasitica. This is the first report of C. parasitica causing canker disease of apple in China.

The Xanthomonas euvesicatoria type III effector XopAU is an active protein kinase that manipulates plant MAP kinase signaling.

The Gram-negative bacterium Xanthomonas euvesicatoria (Xe) is the causal agent of bacterial spot disease of pepper and tomato. Xe delivers effector proteins into host cells through the type III secretion system to promote disease. Here, we show that the Xe effector XopAU, which is conserved in numerous Xanthomonas species, is a catalytically active protein kinase and contributes to the development of disease symptoms in pepper plants. Agrobacterium-mediated expression of XopAU in host and non-host plants activated typical defense responses, including MAP kinase phosphorylation, accumulation of pathogenesis-related (PR) proteins and elicitation of cell death, that were dependent on the kinase activity of the effector. XopAU-mediated cell death was not dependent on early signaling components of effector-triggered immunity and was also observed when the effector was delivered into pepper leaves by Xanthomonas campestris pv. campestris, but not by Xe. Protein-protein interaction studies in yeast and in planta revealed that XopAU physically interacts with components of plant immunity-associated MAP kinase cascades. Remarkably, XopAU directly phosphorylated MKK2 in vitro and enhanced its phosphorylation at multiple sites in planta. Consistent with the notion that MKK2 is a target of XopAU, silencing of the MKK2 homolog or overexpression of the catalytically inactive mutant MKK2K99R in N. benthamiana plants reduced XopAU-mediated cell death and MAPK phosphorylation. Furthermore, yeast co-expressing XopAU and MKK2 displayed reduced growth and this phenotype was dependent on the kinase activity of both proteins. Together, our results support the conclusion that XopAU contributes to Xe disease symptoms in pepper plants and manipulates host MAPK signaling through phosphorylation and activation of MKK2.

Development and validation of a new population-based simulation model of osteoarthritis in New Zealand

To describe the construction and preliminary validation of a new population-based microsimulation model developed to analyse the health and economic burden and cost-effectiveness of treatments for knee osteoarthritis (OA) in New Zealand (NZ). We developed the New Zealand Management of Osteoarthritis (NZ-MOA) model, a discrete-time state-transition microsimulation model of the natural history of radiographic knee OA. In this article, we report on the model structure, derivation of input data, validation of baseline model parameters against external data sources, and validation of model outputs by comparison of the predicted population health loss with previous estimates. The NZ-MOA model simulates both the structural progression of radiographic knee OA and the stochastic development of multiple disease symptoms. Input parameters were sourced from NZ population-based data where possible, and from international sources where NZ-specific data were not available. The predicted distributions of structural OA severity and health utility detriments associated with OA were externally validated against other sources of evidence, and uncertainty resulting from key input parameters was quantified. The resulting lifetime and current population health-loss burden was consistent with estimates of previous studies. The new NZ-MOA model provides reliable estimates of the health loss associated with knee OA in the NZ population. The model structure is suitable for analysis of the effects of a range of potential treatments, and will be used in future work to evaluate the cost-effectiveness of recommended interventions within the NZ healthcare system.

Molecular dissection of distinct symptoms induced by tomato chlorosis virus and tomato yellow leaf curl virus based on comparative transcriptome analysis

The viral infection of plants may cause various physiological symptoms associated with the reprogramming of plant gene expression. However, the molecular mechanisms and associated genes underlying disease symptom development in plants infected with viruses are largely unknown. In this study, we employed RNA sequencing for in-depth molecular characterization of the transcriptional changes associated with the development of distinct symptoms induced by tomato chlorosis virus (ToCV) and tomato yellow leaf curl virus (TYLCV) in tomato. Comparative analysis of differentially expressed genes revealed that ToCV and TYLCV induced distinct transcriptional changes in tomato and resulted in the identification of important genes responsible for the development of symptoms of ToCV (i.e., chlorosis and anthocyanin accumulation) and TYLCV (i.e., yellowing, stunted growth, and leaf curl). Our comprehensive transcriptome analysis can provide molecular strategies to reduce the severity of disease symptoms as well as new insights for the development of virus-resistant crops.

The role of reactive oxygen in the development of Ramularia leaf spot disease in barley seedlings.

Background and Aims Ramularia collo-cygni is an ascomycete fungus that colonizes barley primarily as a benign endophyte, although this interaction can become pathogenic, causing the disease Ramularia leaf spot (RLS). Factors, particularly reactive oxygen species, that resulted in the transition of the fungus from endophyte to necrotrophic parasite and the development of disease symptoms were investigated.MethodsDisease development in artificially inoculated seedlings of barley varieties varying in partial resistance to RLS was related to exposure to abiotic stress prior to inoculation. Histochemical and molecular analysis determined the effect of R. collo-cygni colonization on accumulation of reactive oxygen species and antioxidant gene expression. Development of RLS on barley lines defective in antioxidant enzymes and with altered redox status or non-functional chloroplasts was compared with the accumulation of fungal biomass to determine how these factors affect disease symptom expression.Key ResultsExposure to abiotic stress increased symptom development in all susceptible and most partially resistant barley varieties, in association with greater hydrogen peroxide (H2O2) levels in leaves. Decreased activity of the antioxidant enzymes superoxide dismutase and catalase in transgenic and mutant plants had no effect on the disease transition, whereas manipulation of H2O2 levels during asymptomatic growth of the fungus increased disease symptoms in most susceptible varieties but not in partially resistant plants. Barley mutants that undergo rapid loss of green leaf area when infected by R. collo-cygni or albino mutants with non-functional chloroplasts showed reduced development of RLS symptoms.ConclusionsThese results imply that in seedlings the pathogenic transition of the normally endophytic fungus R. collo-cygni does not result from senescence as such, but rather is promoted by factors that result in changes to host reactive oxygen species. Barley varieties vary in the extent to which these factors promote RLS disease.