Powdery Mildew Spores Research Articles

Nano protective membrane coated wheat to resist powdery mildew.

The plant pathogenic fungus Blumeria graminis f. sp. tritici infects wheat and reduces its yield. The policy of reducing fertilizer and biocide use in sustainable agriculture has prompted researchers to develop more green and efficient management strategies. In this study, a novel nanoprotective membrane (kaolin-nano titanium dioxide-liquid paraffin, referred to as KTP) that could effectively prevent powdery mildew of wheat was prepared by using 1 g/L kaolin, 2 g/L nanotitanium dioxide and 8% (v/v) liquid paraffin. The prevention and control effects of KTP spraying in advance in the pot and field experiments were 98.45% and 83.04%, respectively. More importantly, the weight of 1000 grains of wheat pretreated with KTP was 2.56 g higher than that of wheat infected with powdery mildew, significantly improving wheat yield. KTP delayed the germination of powdery mildew spores on the leaf surface, and inhibited the formation of mycelia. In addition, KTP did not affect the growth of wheat or the survival of earthworms. KTP nanoprotective membrane are a green and safe prevention and control materials that are which is expected to be widely used in agriculture to control wheat powdery mildew.

Detection Method of Fungal Spores Based on Fingerprint Characteristics of Diffraction-Polarization Images.

The most significant aspect of promoting greenhouse productivity is the timely monitoring of disease spores and applying proactive control measures. This paper introduces a method to classify spores of airborne disease in greenhouse crops by using fingerprint characteristics of diffraction-polarized images and machine learning. Initially, a diffraction-polarization imaging system was established, and the diffraction fingerprint images of disease spores were taken in polarization directions of 0°, 45°, 90° and 135°. Subsequently, the diffraction-polarization images were processed, wherein the fingerprint features of the spore diffraction-polarization images were extracted. Finally, a support vector machine (SVM) classification algorithm was used to classify the disease spores. The study's results indicate that the diffraction-polarization imaging system can capture images of disease spores. Different spores all have their own unique diffraction-polarization fingerprint characteristics. The identification rates of tomato gray mold spores, cucumber downy mold spores and cucumber powdery mildew spores were 96.02%, 94.94% and 96.57%, respectively. The average identification rate of spores was 95.85%. This study can provide a research basis for the identification and classification of disease spores.

Study on the dynamic distribution of spores of powdery mildew pathogen in wheat by rotor airflow of plant protection UAV

Plant protection drones are fast and efficient application machines that are characterised by high application efficiency and no damage to crops. They are particularly suitable for small areas of farmland and mountainous terrain in regions such as Asia and are currently the dominant insecticide application technology in China. The presence of wind is a prerequisite for the spread and dissemination of airborne diseases and it can directly influence the distance and height of ascent of pathogenic spores. This paper investigates the effect of downwash airflow generated by the flight altitude of a plant protection drone on the horizontal distribution, vertical distribution and ground distribution of powdery mildew spores in wheat. Monitoring the changing dynamics of airborne powdery mildew conidia using spore traps. The test results show that: the number of powdery mildew pathogenic spores is related to various factors such as weather, relative humidity and wind speed; the release of spores is greatly influenced by airflow disturbances but has little effect at the early stages of sporulation; the disease is caused by the accumulation process of pathogenic spores and in the control of powdery mildew in wheat, preventive spraying should be carried out within 2–3 days of the germination of pathogenic spores. The study lays the foundation for further in-depth research on the spread of powdery mildew spores and improved pest control, and provides a basis for scientific and rational spraying and control by agricultural drones.

A unique effector secreted by Pseudozyma flocculosa mediates its biocontrol activity

BackgroundPseudozyma flocculosa is a highly efficient biocontrol agent (BCA) of powdery mildews whose mode of action remains elusive. It is known to secrete unique effectors during its interaction with powdery mildews but effectors have never been shown to be part of the arsenal of a BCA. Here, we characterize the role of the effector Pf2826 released by Pseudozyma flocculosa during its tripartite interaction with barley and the pathogen fungus Blumeria graminis f. sp. hordei.ResultsWe utilized CRISPR-Cas9-based genome editing and confirmed that secreted P. flocculosa effector Pf2826 is required for full biocontrol activity. We monitored the localization of the effector Pf2826 with C-terminal mCherry tag and found it localized around the haustoria and on powdery mildew spores. His-tagged Pf2826 recombinant protein was expressed, purified, and used as bait in a pull-down assay from total proteins extracted during the tripartite interaction. Potential interactors were identified by LC–MS/MS analysis after removing unspecific interactions found in the negative controls. A two-way yeast two-hybrid assay validated that Pf2826 interacted with barley pathogenesis-related (PR) proteins HvPR1a and chitinase and with an effector protein from powdery mildew.ConclusionsIn contrast to the usual modes of action of competition, parasitism, and antibiosis ascribed to BCAs, this study shows that effector pf2826 plays a vital role in the biocontrol activity of P. flocculosa by interacting with plant PR proteins and a powdery mildew effector, altering the host–pathogen interaction.

Antagonistic Activity and Mechanism of Bacillus subtilis XZ16-1 Suppression of Wheat Powdery Mildew and Growth Promotion of Wheat.

Wheat powdery mildew caused by Blumeria graminis f. sp. tritici (Bgt) is one of the most serious wheat diseases in the world. Biological control is considered an environmentally safe approach to control plant diseases. Here, to develop effective biocontrol agents for controlling wheat powdery mildew, antagonistic strain XZ16-1 was isolated and identified as Bacillus subtilis based on the morphological, biochemical, and physiological characteristics and 16S rDNA sequence. The culture filtrate of B. subtilis XZ16-1 and its extracts had a significant inhibitory effect on the spore germination of Bgt. Moreover, the therapeutic and prevention efficacy of the 100% culture filtrate on wheat powdery mildew reached 81.18 and 83.72%, respectively, which was better than that of chemical fungicide triadimefon. Further antimicrobial mechanism analysis showed that the XZ16-1 culture filtrate could inhibit the development of powdery mildew spores by disrupting the cell membrane integrity, causing reductions in the mitochondrial membrane potential, and inducing the accumulation of reactive oxygen species in the spores. Biochemical detection indicated that XZ16-1 could solubilize phosphate, fix nitrogen, and produce hydrolases, lipopeptides, siderophores, and indole-3-acetic acid. Defense-related enzymes activated in wheat seedlings treated with the culture filtrate indicated that disease resistance was induced in wheat to resist pathogens. Furthermore, a 106 CFU/ml suspension of XZ16-1 increased the height, root length, fresh weight, and dry weight of wheat seedlings by 77.13, 63.46, 76.73, and 19.16%, respectively, and showed good growth-promotion properties. This study investigates the antagonistic activity and reveals the action mechanism of XZ16-1, which can provide an effective microbial agent for controlling wheat powdery mildew.

A novel nanocomposite that effectively prevents powdery mildew infection in wheat

The rapidly growing world population is constantly increasing the demand for food. Being the second most consumed food crop, wheat hold an important economic position. However, powdery mildew is a disease that seriously affects the improvement in the yield and quality of wheat. Currently, triadimefon is the chemical pesticide that is predominantly used to prevent powdery mildew during wheat production. However, using triadimefon not only pollutes the environment, but also deteriorates the quality of harvested wheat grains. In this study, a nanocomposite complex with optimal montmorillonite and dimethyl silicone oil (OMM), which interact with each other through numerous hydrogen bonds. OMM was sprayed onto the surface of the wheat leaves to ensure a uniform nano isolation film that was found to effectively inhibit the contact germination of powdery mildew spores and reduce the disease index by 99.30%. OMM also significantly alleviated both physiological and biochemical stress of powdery mildew infection on the wheat. Furthermore, OMM treatment was found to significantly improve the processed quality of harvested grains. These results demonstrate that OMM treatment is an efficient and environmentally sustainable approach that is suitable for the large-scale prevention of powdery mildew infection in wheat.

Evaluation of mungbean lines for resistance to Cercospora leaves spot and powdery mildew (Erysiphe polygoni) diseases in the green house

Mungbean is an important food crop, it is a source of carbohydrates, protein and B vitamine. Therefore the production must be increased. In increasing the production of mungbean, there are several constrains including disease infection. Leaf spot and powdery mildew are the main diseases on mungbean. Planting the resistant varieties to leaf spot / powdery mildew is an effective, easy, and inexpensive way. The aim of research is to get some resistant mungbean lines against leaf spot and powdery mildew disease. The research was conducted in the greenhouse of the Indonesian Legumes and Tuber Crops Research Institute, from May to September 2019. Evaluation of the resistance of mungbean lines to leaf spot and powdery mildew was carried out consecutively. The material tested was 20 mungbean breeding lines from the breeding section, with one resistant variety and one susceptible variety as a check. Inoculation was done by spraying the suspension of Cercospora cruenta spores of leaf spot, or Erysiphe polygoni spores of powdery mildew, with a density of 104 spores / ml in the afternoon. Observation of the intensity of leaf spot or powdery mildew disease is carried out three times, starting at one week after the first symptoms appeared. The results showed that out of 20 mungbean lines tested, there were no line that were resistant, 7 lines were moderately resistant and 13 lines were moderately susceptible to leaf spot disease. Out of 20 lines of mungbeans tested, 8 lines were resistant and 12 lines were moderately resistant to powdery mildew.

An Easy and Flexible Inoculation Method for Accurately Assessing Powdery Mildew-Infection Phenotypes of Arabidopsis and Other Plants

Reducing crop losses due to fungal diseases requires improved understanding of the mechanisms governing plant immunity and fungal pathogenesis, which in turn requires accurate determination of disease phenotypes of plants upon infection with a particular fungal pathogen. However, accurate disease phenotyping with unculturable biotrophic fungal pathogens such as powdery mildew is not easy to achieve and can be a rate-limiting step of a research project. Here, we have developed a safe, efficient, and easy-to-operate disease phenotyping system using the Arabidopsis-powdery mildew interaction as an example. This system mainly consists of three components: (i) a wooden inoculation box fitted with a removable lid mounted with a stainless steel or nylon mesh of ~50 µm pores for inoculating a flat of plants with fungal spores, (ii) a transparent plastic chamber with a small front opening for minimizing spore escape while conducting inoculation inside, and (iii) a spore-dislodging and distribution method for even and effective inoculation. The protocols described here include the steps and parameters for making the inoculation box and the plastic chamber at a low cost, and a video demonstration of how to use the system to enable even inoculation with powdery mildew spores, thereby improving accuracy and reproducibility of disease phenotyping.

Identification and Analysis of Powdery Mildew Resistance in Melon (Cucumis melo L.) Cultivar Meloni

Background: A powdery mildew-resistant cultivar of melon is needed to increase melon yield crops. Meloni is a superior melon cultivar bred through a crossing between ♀ SL-3 and ♂ PI 371795, resulted by the Laboratory of Genetics and Breeding, Faculty of Biology, UGM. This study aimed to determine the level resistance of Meloni to powdery mildew infection and to identify the powdery mildew species that infected Meloni based on morphological characters. Methods: Meloni seeds were germinated and planted in the greenhouse of PIAT UGM. Powdery mildew spores were inoculated into the leaves after ±2 weeks of age. Leaf infected were scored using the gridline every 3 days for 6 weeks. Scoring results were converted to the diseases index score. Furthermore, powdery mildew species was identified using morphological characters. Results: Meloni had a tolerance level of resistance to the powdery mildew infection. Based on the morphological characters with fibrosin bodies, conidia ovoid-shape and the position of the germ tube in the lateral part of the conidia, powdery mildew that infected Meloni was expected as P. xanthii. Conclusions: Meloni can be expected as an alternative to superior melon seeds resistant to pest and disease infections especially powdery mildew. 

Biological control of powdery mildew on Cornus florida using endophytic Bacillus thuringiensis

Powdery mildew (Erysiphe pulchra) is a devastating disease in nursery production of flowering dogwood (Cornus florida L.). Disease management relies heavily on season-long use of chemical fungicides and there is a demand for alternative strategies. An endophytic bacterium IMC8 was isolated from disease-free C. florida stem tissue, and was shown to be an effective biological control agent against E. pulchra in growth chamber, greenhouse and shadehouse environments. This isolate, identified as Bacillus thuringiensis, exhibited normal growth in NaCl concentrations up to 6%, pH 5 to 11, and temperatures up to 50°C. It was compatible with the conventional fungicide thiophanate methyl and clarified hydrophobic extract of neem seed oil at label recommended concentrations. The isolate IMC8 produced volatile compounds identified by GC/MS, which included antifungal and antibacterial compounds and suggested antibiosis as a mode of action. Scanning electron microscopy revealed lysis of powdery mildew spores and hyphae, which suggested parasitism as a second mode of action. These observations suggested that the isolate IMC8 has potential as a biological control agent in the management of powdery mildew on C. florida, and would tolerate a wide range of salinity, pH and high temperature conditions. It could be included as a component of integrated pest management in rotation or tank mix with lower levels of conventional fungicides, or used with clarified hydrophobic extract of neem seed oil.

The phytopathogen powdery mildew affects food-searching behavior and survival of Coccinella septempunctata

The diet of entomophagous coccinellids is mainly based on aphids and other food sources such as pollen, nectar, or fungal spores. Knowledge of their foraging behavior on plants infected by powdery mildew and their survival on fungal spores is currently limited. In this study, we investigated the olfactory response of Coccinella septempunctata to odor emission of barley plants infected by powdery mildew and their survival on fungal spores in the presence or absence of aphids. Odors released by powdery-mildew infected plants were more attractive for ladybirds compared to those of uninfected controls. After 3 days, the survival rate of ladybirds feeding only on powdery-mildew spores was less than 50%, while for ladybirds feeding exclusively on Rhopalosiphum padi aphids, the survival rate was close to 90%. After 15 days, the highest survival rate (almost 80%) was observed for ladybirds feeding on plants with both aphids and powdery mildew. Molecular analyses confirmed the presence of fungal spores in ladybird guts when feeding either on powdery mildew or on a mixed diet. Our results provide new insights into foraging behavior of entomophagous coccinellids revealing the potential of powdery mildew to be utilized as important non-essential food in a mixed diet, but also its lethal effect if consumed alone.

Host plants and Biological Characteristics of Illeis koebelei Timberlake (Coleoptera: Coccinellidae: Halyziini) in Gyeonggi-do

We investigated mycophagous ladybird, Illeis koebelei from 12 species of plants infected with powdery mildew in Gyeonggi-do, Korea. The pear tree, Pyrus ussuriensis var. macrostipes (Nakai), was most preferred by I. koebelei. This species was found from early July to early November in pear orchards. There was no entomophagous trace in the gut of I. koebelei without powdery mildew spores in a microscope. All stages except egg and pupa are obligate mycophagous, and the feeding potential is ranked as follows: fourth instar, adults, third instar, second instar, and first instar. Feeding amounts of each stage of I. koebelei were 45.6, 144.4, 372.2, 628.1, and 473.7 mm 2 of cucumber powdery mildew per day. Fourth instar larvae showed highest consumption of cucumber powdery mildew. Developmental periods of four larval instars and adults feeding cucumber powdery mildew were 1.2, 2.3, 2.3, 4.6, and 37.7 days, respectively, at 25°C. In this study, we could not determine the feeding potential of I. koebelei against the cucumber powdery mildew; therefore, and further studies are required to elucidate the potential of this species as a biological control agent, e.g., mass rearing, selection of low toxic chemical agents for Integrated Pest Management (IPM), and control techniques against powdery mildew in agro-ecosystems.

Development of Powdery Mildew Fungus on Cucumber Leaves Acclimatized to Different CO2 Concentrations

The present study evaluated the development of powdery mildew fungus (Podosphaera xanthii) on leaves of cucumber (Cucumis sativus L.) acclimatized to different CO2 concentrations ([CO2]) to examine plant–pathogen interactions under the wide range of [CO2] that can occur in greenhouse cultivation. Seedlings of resistant and nonresistant cultivars were acclimatized to reduced (200 µmol·mol−1), ambient (400 µmol·mol−1), or elevated (1000 µmol·mol−1) [CO2]. Powdery mildew spores were inoculated onto the adaxial surface of cotyledons or first true leaves, and colonization was measured after 7 days. Colony density decreased as acclimatization [CO2] increased at the cotyledon stage but increased at the first-true-leaf stage in both cultivars. This result implies that when the effects of [CO2] on plant–pathogen interactions are described, growing stage must be specified. The acclimatization [CO2] was correlated positively with leaf mass per area, dry matter content, and carbon (C) content and negatively with nitrogen (N) content at both stages. Therefore, these leaf properties could not explain the changes in host-plant susceptibility between stages. The effect of acclimatization [CO2] was greater on the resistant cultivar than on the nonresistant cultivar, indicating that the resistant cultivar was more responsive.

The Cytokinin-Activated Transcription Factor ARR2 Promotes Plant Immunity via TGA3/NPR1-Dependent Salicylic Acid Signaling in Arabidopsis

Cytokinins affect plant immunity to various pathogens; however, the mechanisms coupling plant-derived cytokinins to pathogen responses have been elusive. Here, we found that plant-derived cytokinins promote resistance of Arabidopsis to Pseudomonas syringae pv. tomato DC3000 (Pst). Modulated cytokinin levels or signaling activity in CKX- or IPT-overexpressing plants or in ahk2 ahk3 mutants correlated with altered resistance. In fact, the cytokinin-activated transcription factor ARR2 contributes specifically to Pst resistance. The salicylic acid (SA) response factor TGA3 binds ARR2, and mutation of TGA-binding cis-elements in the Pr1 promoter abolished cytokinin- and ARR2-dependent Pr1 activation. Cytokinin treatment did not increase pathogen resistance in tga3 plants, as the cytokinin-dependent induction of Pr1 was eliminated. Moreover, SA signaling enhanced binding of ARR2/TGA3 to the Pr1 promoter. Taken together, these results show that cytokinins modulate the SA signaling to augment resistance against Pst, a process in which the interaction between TGA3 and ARR2 is important.

Potential Sources of Resistance to Cucurbit Powdery Mildew in U.S. Plant Introductions of Bottle Gourd

Powdery mildew (Podosphaera xanthii) can cause severe damage to cucurbit crops grown in open fields and greenhouses. In recent years, there has been an increased interest in the United States in grafting watermelon plants onto various cucurbit rootstocks. Bottle gourd plants (Lagenaria siceraria) are being used throughout the world as rootstocks for grafting watermelon. Although gourd plants are beneficial, they may still be susceptible hosts to various soilborne and foliar diseases. Bottle gourd plant introductions (PI) resistant to diseases and pests can be a valuable source of germplasm in rootstock breeding programs. We evaluated 234 U.S. PIs of L. siceraria for tolerance to powdery mildew in two greenhouse tests. Young seedlings were inoculated by dusting powdery mildew spores of melon race 1 on the cotyledons. Plants were rated 2 weeks after inoculation using a 1 to 9 scale of increasing disease severity. Although none of the L. siceraria PIs were immune to powdery mildew, several PIs had significantly lower levels of powdery mildew severity compared with susceptible watermelon cultivar Mickey Lee. The experiment was repeated with 26 select PIs on whole seedlings and cotyledon disks. Significant variability in the level of resistance to powdery mildew on plants within PI was observed. Moderate resistance in several PIs to powdery mildew was confirmed. PI 271353 had consistently lower ratings in the various tests and can be considered the most resistant to P. xanthii race 1 among the L. siceraria accessions evaluated in this study. A few other PIs with moderate resistance to powdery mildew included PI 271357, PI 381840, and PI 273663. These results suggest that novel sources of resistance could be developed by careful selection and screening of several of the PIs with moderate resistance described in our study.

Evaluation of Lecanicillium longisporum, Vertalec ® for simultaneous suppression of cotton aphid, Aphis gossypii, and cucumber powdery mildew, Sphaerotheca fuliginea, on potted cucumbers

The commercial preparation of Lecanicillium longisporum, Vertalec ® was evaluated for simultaneous suppression of cotton aphid and cucumber powdery mildew on potted cucumber plants. Vertalec was applied onto cucumber plants that had been infested with either cotton aphid, spores of Sphaerotheca fuliginea or both. Irradiation-inactivated Vertalec (II Vertalec) was also applied to an identical series of cucumber plants as a control. The Vertalec was highly pathogenic against adult aphids with an LT 50 of 6.9 days. II Vertalec did not affect aphid survival. Application of either active or II Vertalec significantly suppressed spore production of S. fuliginea compared to the water control. For dual control assays, Vertalec applications were made one day after infestation of both aphid and S. fuliginea onto potted cucumbers. Fifteen days after the Vertalec treatments, the numbers of surviving aphids and the production of powdery mildew spores were significantly reduced compared with the water control. The presence of aphids also suppressed S. fuliginea spore production. Our results suggest the potential of a dual role for Vertalec as a microbial control agent of aphids and powdery mildew in cucumber.

Cloning of a novel phosphateserine aminotransferase gene from a Triticum aestivum-Elytrigia elongatum alien substitution line with resistance to powdery mildew

Shannong 551, a T. aestivum-E. elongatum alien substitution line with resistance to powdery mildew, was inoculated with pathogenic spores of powdery mildew. The leaf samples were prepared 48 h after inoculation for scanning electron microscopy. The result showed that germination of spores and growth of young mycelia on leaves of Shannong 551 were suppressed at the early stage of infection. At the same time, RNAs were prepared from the leaves for the cloning of WRP1 and RPW2 by cDNA RDA and RACE technology. BLAST analysis of the sequences indicated that both WRP1 and RPW2 were novel genes. WRP1 contains no complete ORF. RPW2 contains the conserved structure domain of aminotransferase, and its DNA sequence shares high homology with genes of phosphateserine aminotransferase in many organisms. Therefore, it is speculated as a novel phosphateserine aminotransferase gene. The results of Northern blot suggested that expression of RPW2 occurred at the early stage of infection by powdery mildew. Southern blot using the probe of RPW2, in which there was strong hybridizing signals in both genome of Shannong 551 and E. elongatum, but not in those of Jinan 13 and Lumai No.5, indicated that RPW2 derived from the genome of E. elongatum.

Fatty acids with certain structural characteristics are potent inhibitors of germination and inducers of cell death of powdery mildew spores

Fatty acids with certain structural characteristics inhibited the germination of Erysiphe polygoni spores. Inhibition of germination was associated with disruption of cellular organization, which included rupture of vacuoles, loss of structural order and condensation of cellular contents and finally considerable shrinkage of the spores. These events could be invoked at a low concentration, i.e. 20μM, of the fatty acids and were irreversible after 15min. Cellular disorganization was detected in a relatively short time, approx. 1h. In examining a group of fatty acids with different structural characteristics, it was found that the presence of a cis double bond was important for the inhibitory activities. Saturated and trans -monounsaturated fatty acids were ineffective. Also, at 20μM, polyunsaturated fatty acids had marginal activities and not all cis -monounsaturated fatty acids were inhibitory. It was noted that the fatty acids with the highest activities shared a common feature,. i.e. a double bond 6 or 7 carbons from either end of the carbon chain. The results suggest that the presence of a Δ6, Δ7, ω6, or ω7 double bond by the cis -monounsaturated fatty acids is an important characteristic for the potent inhibitory activity. In contrast, the presence of an acid group was apparently not a critical requirement for activity, because twocis -monounsaturated fatty alcohols tested also exhibited similar potent inhibitory and disruptive effects.Palmitoleic acid-induced spore shrinkage could be partially prevented by BaCl2 and fully prevented by spermine. These are compounds known to block certain K+ channels, suggesting the involvement of K+ channels in palmitoleic acid effects. Also, when the spores were stained with the K+ binding ratiometric dye, tetra (acetoxymethyl) ester benzofuran isophthalate (PBFI-AM), disappearance of fluorescence at 340nm was observed in spores treated with palmitoleic acid, indicating a loss of K+ in these spores.

Relationship between cutin monomers and tomato resistance to powdery mildew infection

Introduction of the yeast Δ-9 desaturase gene into tomato plants (Lycopersicon esculentum Mill.) led to distinctive changes in their fatty acid and cutin monomer profiles. Most evident were the dramatic increases in 9-hexadecenoic acid and similarly sharp increases of this acid as a cutin monomer in transgenic plants. The transgenic plants were also more resistant to powdery mildew caused by Erysiphe polygoni DC. Cutin monomers from transgenic plants inhibited the germination of spores of powdery mildew. Germination of spores on transgenic cutin sheets was also inhibited.

Barley coleoptile peroxidases. Purification, molecular cloning, and induction by pathogens.

A cDNA clone encoding the Prx7 peroxidase from barley (Hordeum vulgare L.) predicted a 341-amino acid protein with a molecular weight of 36,515. N- and C-terminal putative signal peptides were present, suggesting a vacuolar location of the peroxidase. Immunoblotting and reverse-transcriptase polymerase chain reaction showed that the Prx7 protein and mRNA accumulated abundantly in barley coleoptiles and in leaf epidermis inoculated with powdery mildew fungus (Blumeria graminis). Two isoperoxidases with isoelectric points of 9.3 and 7.3 (P9.3 and P7.3, respectively) were purified to homogeneity from barley coleoptiles. P9.3 and P7.3 had Reinheitszahl values of 3.31 and 2.85 and specific activities (with 2,2'-azino-di-[3-ethyl-benzothiazoline-6-sulfonic acid], pH 5.5, as the substrate) of 11 and 79 units/mg, respectively. N-terminal amino acid sequencing and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry peptide analysis identified the P9. 3 peroxidase activity as due to Prx7. Tissue and subcellular accumulation of Prx7 was studied using activity-stained isoelectric focusing gels and immunoblotting. The peroxidase activity due to Prx7 accumulated in barley leaves 24 h after inoculation with powdery mildew spores or by wounding of epidermal cells. Prx7 accumulated predominantly in the epidermis, apparently in the vacuole, and appeared to be the only pathogen-induced vacuolar peroxidase expressed in barley tissues. The data presented here suggest that Prx7 is responsible for the biosynthesis of antifungal compounds known as hordatines, which accumulate abundantly in barley coleoptiles.