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Agriculture is a source of living in many areas and acts a backbone for the Indian economy. Agriculture crops are affected by fungus and bacteria to cause diseases in the plant. Brown spot is a serious hazard that takes place on leaves of the rice plant. The presence of fungus ‘Cochliobolus miyabeanus’ is the main factor for brown spot disease. The severity of the disease depends on growth of the fungus. The development of fungus depends on several factors such as temperature, humidity and rainfall. Whenever the disease occurs on a plant, level of the infection plays an importance in crop yield. Most of the farmer follow books or use their experience to detect the disease in rice plants which is a time-consuming process and requires lot of attention to produce more rice yield is a challenging task. Fuzzy logic is identified as powerful tool for disease detection. In this paper, fuzzy logic system is proposed to determine the level of presence of Cochliobolus miyabeanus and alert the farmer to take the primitive steps.

Because pathogens use diverse infection strategies, plants cannot use one-size-fits-all defence and modulate defence responses based on the nature of pathogens and pathogenicity mechanism. Here, we report that a rice glycoside hydrolase (GH) plays contrasting roles in defence depending on whether a pathogen is hemibiotrophic or necrotrophic. The Arabidopsis thaliana MORE1 (Magnaporthe oryzae resistance 1) gene, encoding a member of the GH10 family, is needed for resistance against M. oryzae and Alternaria brassicicola, a fungal pathogen infecting A. thaliana as a necrotroph. Among 13 rice genes homologous to MORE1, 11 genes were induced during the biotrophic or necrotrophic stage of infection by M. oryzae. CRISPR/Cas9-assisted disruption of one of them (OsMORE1a) enhanced resistance against hemibiotrophic pathogens M. oryzae and Xanthomonas oryzae pv. oryzae but increased susceptibility to Cochliobolus miyabeanus, a necrotrophic fungus, suggesting that OsMORE1a acts as a double-edged sword depending on the mode of infection (hemibiotrophic vs. necrotrophic). We characterized molecular and cellular changes caused by the loss of MORE1 and OsMORE1a to understand how these genes participate in modulating defence responses. Although the underlying mechanism of action remains unknown, both genes appear to affect the expression of many defence-related genes. Expression patterns of the GH10 family genes in A. thaliana and rice suggest that other members also participate in pathogen defence.

The novel trichothecene 12-deoxytrichodermin (3) was isolated from the fungus Trichoderma sp. 1212-03, and included with other known natural trichothecenes in a structure-activity relationship investigation against a human colon cancer cell line (COLO201) and filamentous fungus Cochliobolus miyabeanus. This revealed that the 12-epoxide functionality is critical for the cytotoxicity of simple trichothecenes trichodermin (4) and deoxynivalenol (2), while not critical for the cytotoxicity of roridin J (6) and epiisororidin E (8). In contrast, 12-epoxide is essential for the antifungal activity.

Waitea circinata (Warcup & Talbot) is an orchid antagonist mycorrhizal fungus with biocontrol potential against rice pathogens. This study aimed to optimize the extraction method, obtain a new extract and evaluate its efficiency against rice pathogens in vitro and in vivo, as well as to compare it with other extraction methods and W. circinata. The extracts were obtained and screened for in vitro growth inhibition against the pathogens Cochliobolus miyabeanus, Monographella albescens and Sarocladium oryzae, using the following extracts: mycelial, crude, lyophilized and mycelial mass. An additional in vitro assay was performed with the principal rice pathogen (Magnaporthe oryzae), in order to evaluate the conidial germination and appressorium formation. Based on this evaluation, the lyophilized and mycelial mass extracts were tested in vivo against rice blast (M. oryzae) and compared to the W. circinata mycelial suspension, in different application forms (simultaneous and previous). The mycelial mass extract inhibited all the pathogens, and the crude and lyophilized extracts inhibited C. miyabeanus and M. albescens, respectively. The mycelial mass extract inhibited the M. oryzae conidial germination and appressorium formation by 80 %, and the simultaneous and previous applications suppressed the rice blast by 94 %. These results indicate that the new extract can be used to control rice pathogens. RESUMO Waitea circinata (Warcup & Talbot) é um fungo antagonista micorrízico de orquídea, com potencial de biocontrole contra patógenos de arroz. Objetivou-se otimizar o método de extração, obter um novo extrato e avaliar sua eficiência contra patógenos de arroz in vitro e in vivo, bem como compará-lo com outros métodos de extração e W. circinata. Os extratos foram obtidos e testados para inibição de crescimento in vitro contra os patógenos Cochliobolus miyabeanus, Monographella albescens e Sarocladium oryzae, utilizando-se os seguintes extratos: micelial, bruto, liofilizado e de massa micelial. Um ensaio adicional in vitro foi realizado com o principal patógeno do arroz (Magnaporthe oryzae), para avaliar a germinação de conídios e a formação de apressórios. Com base nessa avaliação, os extratos liofilizado e de massa micelial foram testados in vivo contra a brusone do arroz (M. oryzae) e comparados com a suspensão micelial de W. circinata, em diferentes formas de aplicação (simultânea e prévia). O extrato de massa micelial inibiu todos os patógenos, e os extratos bruto e liofilizado inibiram C. miyabeanus e M. albescens, respectivamente. O extrato de massa micelial inibiu a germinação de conídios e a formação de apressórios de M. oryzae em 80 %, e as aplicações simultâneas e prévias suprimiram a brusone em 94 %. Os resultados indicam que o novo extrato pode ser usado para controlar patógenos de arroz.

Pathogens utilize multiple types of effectors to modulate plant immunity. Although many apoplastic and cytoplasmic effectors have been reported, nuclear effectors have not been well characterized in fungal pathogens. Here, we characterize two nuclear effectors of the rice blast pathogen Magnaporthe oryzae. Both nuclear effectors are secreted via the biotrophic interfacial complex, translocated into the nuclei of initially penetrated and surrounding cells, and reprogram the expression of immunity-associated genes by binding on effector binding elements in rice. Their expression in transgenic rice causes ambivalent immunity: increased susceptibility to M. oryzae and Xanthomonas oryzae pv. oryzae, hemibiotrophic pathogens, but enhanced resistance to Cochliobolus miyabeanus, a necrotrophic pathogen. Our findings help remedy a significant knowledge deficiency in the mechanism of M. oryzae–rice interactions and underscore how effector-mediated manipulation of plant immunity by one pathogen may also affect the disease severity by other pathogens., Plant pathogens secrete various effectors to manipulate host immunity. Here, Kim et al. describe two Magnaporthe oryzae effectors that translocate into the nuclei of infected rice cells and reprogram expression of immunity-associated genes, increasing susceptibility to hemibiotrophic pathogens.

The panicle branch, which is the key node for transport of photosynthesis products from source to sink, is vulnerable to many diseases caused by fungal pathogens, such as Magnaporthe oryzae, Cochliobolus miyabeanus. Among these diseases, rice blast is the most important one which causes devastating losses in many regions. In 2019 and 2020, panicle branch rot of rice with a symptom which could be mistaken with rice blast was observed in a paddy field, where is not traditional epidemical region of rice blast, in Fuyang, Zhejiang province. In 2020, similar symptom was also observed in Hubei and Anhui Province. In a paddy field in Fuyang, the symptom appeared on more than 30% investigated panicles. Diseased panicle exhibited brown to black lesions on primary or secondary branches as well as pedicels, however the grain and the neck of spike could not be infected which is the most obvious difference with rice blast. Obviously, the disease can't destroy the entire function of branch and blank grain was rarely observed, so its damage is not comparable with neck blast. Normally, it caused incomplete grain filing commonly leading to 5% - 25% grain weight loss. During the booting stage of rice, local solar irradiation time and temperature were fewer and lower than common years which may be responsible for losses caused by this disease. After surface sterilized, lesion parts cut from infected branches from 25 panicle samples were cultured on 2% water agar at 28℃ for 24-28 h, and fungi were isolated and purified by mycelial tip transferring. Among 31 isolates, 26 showed similar cultural characters. The wool-like mycelia were luxuriant and grew rapidly on PDA spreading the whole 9 cm petri dish in less than a week at 28 ℃. The mycelia were white to ashen at beginning and gradually turned black from center of the plate after 5 days culture at 28 ℃. Hyphae were smooth, branched, septate, hyaline or pale brown. Conidia were single-celled, black, spherical to subspherical, and 10.2 to 14.6 × 12.2 to 15.7 μm (n=50) in dimension and born on tip of hyaline and ampulliform conidiophores. The fungus showed similar morphological characteristics with Nigrospora oryzae (1). ITS sequences of 6 representative strains of the fungus were amplified, sequenced with primer pair, ITS1/ITS4 (2), and submitted into GenBank with an accession number, MW228165. Phylogenetic analysis was conducted with sequences of reference strains (3). The result showed that the fungus obtained in this study was fallen into the same group with N.oryzae. In view of above both morphological and molecular analysis, the strains were finally identified as N. oryzae. Pathogenicity tests were conducted in triplicate with rice panicles in initial heading stage. Fifty panicles were wounded on branches with needles and inoculated by spreading the conidia suspension (10μl, 1 × 106 conidia ml-1) on the wounds. The panicles used as control were treated in same way with 10μl of sterile water. The inoculated and control plants were kept in dark, 25 ℃ and relative humidity of more than 85% for 24 h in culture chamber. Symptoms appeared on 44 of 50 inoculated panicles which were basically similar with those observed in paddy field, while negative controls remained symptomless. The fungi re-isolated from inoculated panicles were also confirmed as N. oryzae by both morphological and molecular analysis. To the best of our knowledge, this is the first report of N. oryzae causing panicle branch rot disease on Oryza sativa (rice). This disease not only cause yield losses and lower milling quality, but could also be mistaken as rice blast incurring unnecessary fungicides spray.

Bipolaris oryzae causes rice brown spot (RBS) in common rice (Oryza sativa) inflicting substantial grain yield losses worldwide. Knowledge of the population structure, genetic diversity, and sexual recombination of the fungal pathogen can help to implement effective disease management strategies. In this study, B. oryzae isolates sampled from Iran, The Philippines, and Japan were analyzed with 12 simple-sequence repeat (SSR) markers newly developed from the genome sequence of the fungus. Among the 288 B. oryzae isolates genotyped, 278 unique haplotypes were identified. High genotype numbers (richness) with even distribution (evenness) were found within the collection sites. Both mating types, MAT1-1 and MAT1-2, were present in each collection area, and the sexual state was induced under controlled conditions with production of viable ascospores. However, the tests of linkage disequilibrium did allow rejection of the hypothesis of random mating. Discriminant analysis of principal components (DAPC) revealed that the B. oryzae collection formed three clusters each consisting of isolates from different collection sites. Analysis of molecular variance (AMOVA) showed that genetic variation among clusters was 18.7%, with the rest distributed within clusters (Rst = 0.187, P < 0.001). Statistically significant pairwise-genetic differentiation was found between the clusters. These results show that Asian B. oryzae isolates are genetically diverse, and overall distributed in three groups. Those findings will be helpful in managing the disease and guide the use of representative set of isolates needed for selection of RBS resistant rice varieties. This article is protected by copyright. All rights reserved.

The fungal genus Cochliobolus describes necrotrophic pathogens that give rise to significant losses on rice, wheat, and maize. Revealing plant mechanisms of non-host resistance (NHR) against Cochliobolus will help to uncover strategies that can be exploited in engineered cereals. Therefore, we developed a heterogeneous pathosystem and studied the ability of Cochliobolus to infect dicotyledons. We report here that C. miyabeanus and C. heterostrophus infect Arabidopsis accessions and produce functional conidia, thereby demonstrating the ability to accept Brassica spp. as host plants. Some ecotypes exhibited a high susceptibility, whereas others hindered the necrotrophic disease progression of the Cochliobolus strains. Natural variation in NHR among the tested Arabidopsis accessions can advance the identification of genetic loci that prime the plant's defence repertoire. We found that applied phytotoxin-containing conidial fluid extracts of C. miyabeanus caused necrotic lesions on rice leaves but provoked only minor irritations on Arabidopsis. This result implies that C. miyabeanus phytotoxins are insufficiently adapted to promote dicot colonization, which corresponds to a retarded infection progression. Previous studies on rice demonstrated that ethylene (ET) promotes C. miyabeanus infection, whereas salicylic acid (SA) and jasmonic acid (JA) exert a minor function. However, in Arabidopsis, we revealed that the genetic disruption of the ET and JA signalling pathways compromises basal resistance against Cochliobolus, whereas SA biosynthesis mutants showed a reduced susceptibility. Our results refer to the synergistic action of ET/JA and indicate distinct defence systems between Arabidopsis and rice to confine Cochliobolus propagation. Moreover, this heterogeneous pathosystem may help to reveal mechanisms of NHR and associated defensive genes against Cochliobolus infection.

Rice (Oryza sativa L.) is an important cereal crop in Venezuela and is planted on approximately 200,000 ha. Fungal diseases caused by the Helminthosporium complex of fungi are considered a major constraint to rice production. In 2005, a blotter method was used to identify fungi associated with rice seed. Seeds (150) of cv. INIA-017 were placed on a plate with moistened filter paper and incubated at 26 ± 2°C with a 12-h light/12-h dark cycle for 5 days until fungal sporulation. Single spores were transferred to 2% water agar (WA). Germinated spores were then transferred to potato dextrose agar (PDA), and resultant colonies were preserved until used. To induce sporulation, 4-mm-diameter discs were cut from 72-h-old cultures, transferred to WA, and incubated at 26 ± 2°C for 48 h (modified technique of Alcorn [1]). Discolored seed yielded isolates of Bipolaris oryzae (Berda de Haan) Shoemaker and Exserohilum rostratum (Drechs.) Leonard and Suggs (2). Pathogenicity tests were performed on 60-day-old rice plants of cv. Blue Bonnet 50. Four plants per pot were used for each isolate. Before inoculation, plants were placed in humidity chambers for 48 h. Spores were harvested from discs into a beaker containing 50 mL of a dilute gelatin solution (Gelatin Difco; 5 g/100 mL of sterile distilled water [SDW]). Spore suspensions were filtered through cheesecloth and adjusted to 7 × 104 spores per ml. The inoculated plants were placed in humidity chambers for 72 h. Control plants were sprayed with SDW. Inoculated plants were removed from humidity chambers and placed on a greenhouse bench. Plants inoculated with B. oryzae were symptomatic 5 days after inoculation; resultant lesions were red-brown with chlorotic borders. Lesions subsequently turned gray with reddish borders and were rhombic in shape. Plants inoculated with E. rotratum were symptomatic 7 days after inoculation with elliptical lesions that were orange-brown along venation. These lesions turned gray with reddish borders. Both fungi were reisolated from symptomatic leaf tissue. To our knowledge, this is the first report of E. rostratum from rice in Venezuela. References: (1) J. L. Alcorn. Mycotaxon. 17:1, 1983. (2) A. Sivanesam. Mycol. Pap. 158, 1987.

The Cochliobolus genus consist of over 55 species among which the 5 most devastating are Cochliobolus carbonum, Cochliobolus heterostrophus, Cochliobolus miyabeanus, Crocus sativus and Cochliobolus lunatus causing damages in sorghum, wheat, rice, maize, cassava and soybean estimated at over 10 billion USD per annum worldwide. The dynamic pathogenicity of Cochliobolus species and the plethora of infected hosts is determined by the evolution of virulence determinants such as the velvet-like B protein (VelB). Nonetheless, the knowledge on the distribution of Cochliobolus VelB and its implication in pathogenicity and fungicide resistance are often lacking. By scanning through the annotated genomes of C. lunatus, C. heterostrophus, C. carbonum, C. victoriae, C. sativus and C. miyabeanus, it is revealed that the numbers of ortholog VelB and cognates vary. By using the phylogenetic approach, it is established that the diversification rates among velvet-domain-containing proteins for phytopathogenic Cochliobolus species could impact differently on their oxidant and fungicide resistance potentials, ability to form appressoria-like structures and infection pegs during infection. This study provides new insights into the pathogenicity evolution of Cochliobolus species at the VelB locus which is relevant for designing effective strategies for durable management of Cochliobolus diseases.

Broad-spectrum disease resistance against two or more types of pathogen species is desirable for crop improvement. In rice, Xanthomonas oryzae pv. oryzae (Xoo), the causal bacteria of rice leaf blight, and Magnaporthe oryzae, the fungal pathogen causing rice blast, are two of the most devastating pathogens. We identified the rice BROAD-SPECTRUM RESISTANCE 1 (BSR1) gene for a BIK1-like receptor-like cytoplasmic kinase using the FOX hunting system, and demonstrated that BSR1-overexpressing (OX) rice showed strong resistance to the bacterial pathogen, Xoo and the fungal pathogen, M. oryzae. Here, we report that BSR1-OX rice showed extended resistance against two other different races of Xoo, and to at least one other race of M. oryzae. In addition, the rice showed resistance to another bacterial species, Burkholderia glumae, which causes bacterial seedling rot and bacterial grain rot, and to Cochliobolus miyabeanus, another fungal species causing brown spot. Furthermore, BSR1-OX rice showed slight resistance to rice stripe disease, a major viral disease caused by rice stripe virus. Thus, we demonstrated that BSR1-OX rice shows remarkable broad-spectrum resistance to at least two major bacterial species and two major fungal species, and slight resistance to one viral pathogen.

Rice plants accumulate various specialized metabolites, including phenylamides, in response to pathogen attack. We prepared 25 phenylamides, and developed a method of analyzing them by multiple reaction monitoring with liquid chromatography coupled with tandem mass spectrometry. We analyzed phenylamides in rice leaves infected with Cochliobolus miyabeanus and Xanthomonas oryzae. The phenylamides induced included benzoyltryptamine, cinnamoyl-, p-coumaroyl-, feruloyl-, and benzoylserotonins, cinnamoyl and benzoyltyramines, feruloylagmatine, and feruloylputrescine. Some of the phenylamides exhibited antimicrobial activity against C. miyabeanus and X. oryzae, indicating that they are phytoalexins. Treatment with jasmonic acid, salicylic acid, 6-benzylaminopurine, and ethephone also induced phenylamide accumulation. The compositions of the induced amides varied depending on the plant hormone used, and cinnamoyltryptamine, cinnamoylserotonin, and cinnamoyltyramine were not induced by the plant hormones. These findings suggest that several plant hormones and additional factors are involved in phenylamide accumulation in response to pathogen infection in rice. Rice plant accumulates phenylamides in response to infection, wounding and treatments with jasmonic acid, salycilic acid, cytokinin, and ethylene.

Cochliobolus miyabeanus is known as a significant causal agent in relation to brown spot disease of rice and causes significant yield losses. In the present study, the complete mitochondrial genome was determined using next-generation sequencing technology. This complete mitogenome is a circular molecule of 124,887 bp in length. It contains 13 conserved protein-coding genes, 21 transfer RNA genes, 2 ribosomal RNA genes and 9 open reading frames. The overall base composition of C. miyabeanus is 35.4% A, 34.4% T, 14.4% C, 15.8% G, with a CG content of 30.2%. Phylogenetic analysis based on concatenated protein genes from 15 taxa within Pezizomycotina showed that C. miyabeanus is closely related to Bipolaris cookei in the family Pleosporaceae (Dothideomycetes, Pleosporales). This work would facilitate the understanding of systematics and evolutionary biology of phytopathogenic fungi.

Summary Over recent decades, a multitude of studies have shown the ability of silicon (Si) to protect various plants against a range of microbial pathogens exhibiting different lifestyles and infection strategies. Despite this relative wealth of knowledge, an understanding of the action mechanism of Si is still in its infancy, which hinders its widespread application for agricultural purposes. In an attempt to further elucidate the molecular underpinnings of Si-induced disease resistance, we studied the transcriptome of control and Si-treated rice plants infected with the necrotrophic brown spot fungus Cochliobolus miyabeanus. Analysis of brown spot-infected control plants suggested that C. miyabeanus represses plant photosynthetic processes and nitrate reduction in order to trigger premature senescence and cause disease. In Si-treated plants, however, these pathogen-induced metabolic alterations are strongly impaired, suggesting that Si alleviates stress imposed by the pathogen. Interestingly, Si also significantly increased photorespiration rates in brown spot-infected plants. Although photorespiration is often considered as a wasteful process, recent studies have indicated that this metabolic bypass also enhances resistance during abiotic stress and pathogen attack by protecting the plant's photosynthetic machinery. In view of these findings, our results favour a scenario in which Si enhances brown spot resistance by counteracting C. miyabeanus-induced senescence and cell death via increased photorespiration. Moreover, our results shed light onto the mechanistic basis of Si-induced disease control and support the view that, in addition to activating plant immune responses, Si can also reduce disease severity by interfering with pathogen virulence strategies.

The Pseudomonas- derived cyclic lipopeptide orfamide can induce resistance to Cochliobolus miyabeanus but not to Magnaporthe oryzae in rice. Abscisic acid signaling is involved in the induced systemic resistance response triggered by orfamide. Diverse natural products produced by beneficial Pseudomonas species have the potential to trigger induced systemic resistance (ISR) in plants, and thus may contribute to control of diseases in crops. Some beneficial Pseudomonas spp. can produce cyclic lipopeptides (CLPs), amphiphilic molecules composed of a fatty acid tail linked to an oligopeptide which is cyclized. CLPs can have versatile biological functions, but the capacity of Pseudomonas-derived CLPs in triggering ISR responses has barely been studied. Pseudomonas protegens and related species can produce orfamide-type CLPs. Here we show that in rice, orfamides can act as ISR elicitors against the necrotrophic fungus Cochliobolus miyabeanus, the causal agent of brown spot disease, but are not active against the blast fungus Magnaporthe oryzae. Orfamide A can trigger early defensive events and activate transcripts of defense-related genes in rice cell suspension cultures, but does not cause cell death. Further testing in rice cell suspension cultures and rice plants showed that abscisic acid signaling, the transcriptional activator OsWRKY4 and pathogenesis-related protein PR1b are triggered by orfamide A and may play a role in the ISR response against C. miyabeanus.

The integrated management of rice leaf blast (Magnaporthe oryzae) is carried out mainly with the adoption of chemical control. However, the search for alternative practices has grown in recent years. Thus, the variability of 28 Sarocladium oryzae isolates was evaluated for the cerulenin production, as well as its potential for reducing the severity of rice leaf blast, quantifying the activity of enzymes linked to the plant defense mechanisms. More than 55 % of the S. oryzae isolates were antagonistic to the pathogens M. oryzae, Cochliobolus miyabeanus, Thanatephorus cucumeris and Monographella albescens, and 60 % of the isolates produced cerulenin at detectable levels. Both BRM 6461 (296.0 µg mL-1) and BRM 6493 (undetectable cerulenin) inhibited the formation of M. oryzae appressoria in 89.5 % and 85 %, respectively. The BRM 6461 isolate, applied as conidial suspension and filtered, reduced the severity of rice leaf blast in 68.8 % and 75.5 %, respectively. The enzymatic activity in the presence of M. oryzae was higher for lipoxygenase at 5 h (filtered) and at 24 h and 72 h (conidial suspension) after the pathogen inoculation. For phenylalanine ammonia lyase, the highest expression was at 5 h (filtered) and 72 h (conidial suspension). The enzymes chitinase, β-1,3-glucanase and peroxidase and the salicylic acid phytohormone presented no differences, in relation to the controls (water and M. oryzae). The filtered from the BRM 6461 isolate, basically constituted by cerulenin, reduced the severity of rice leaf blast and possibly activated the defense mechanisms of the rice plants against M. oryzae. RESUMO O manejo integrado de brusone (Magnaporthe oryzae) em arroz é realizado, principalmente, com a adoção de controle químico. No entanto, a busca por práticas alternativas tem crescido nos últimos anos. Assim, avaliou-se a variabilidade de 28 isolados de Sarocladium oryzae, quanto à produção de cerulenina, e o seu potencial na redução da severidade de brusone foliar em arroz, quantificando-se a atividade de enzimas ligadas ao mecanismo de defesa das plantas. Mais de 55 % dos isolados de S. oryzae foram antagônicos aos patógenos M. oryzae, Cochliobolus miyabeanus, Thanatephorus cucumeris e Monographella albescens, e 60 % dos isolados produziram cerulenina em níveis detectáveis. Tanto BRM 6461 (296,0 µg mL-1) quanto BRM 6493 (cerulenina não detectável) inibiram a formação de apressórios de M. oryzae em 89,5 % e 85 %, respectivamente. O isolado BRM 6461, aplicado na forma de suspensão de conídios e filtrado, reduziu a severidade da brusone em 68,8 % e 75,5 %, respectivamente. A atividade enzimática, na presença de M. oryzae, foi maior para lipoxigenase após 5 h (filtrado) e 24 h e 72 h (suspensão de conídios) da inoculação do patógeno. Para fenilalanina-amônia liase, a maior expressão ocorreu após 5 h (filtrado) e 72 h (suspensão de conídios). As enzimas quitinase, β-1,3-glucanase e peroxidase e o fitohormônio ácido salicílico não apresentaram diferenças em relação aos controles (água e M. oryzae). O filtrado do isolado BRM 6461, constituído basicamente por cerulenina, reduziu a severidade da brusone e, possivelmente, ativou os mecanismos de defesa da planta de arroz contra M. oryzae.

The effects of essential oils (EO), cold water (CWE), hot water (HWE) and ethanol (ETHE) extracts of Callistemon citrinus L. and Cymbopogon citratus (DC) Stapf on the radial growth of Alternaria padwickii (Ganguly) M.B. Ellis and Bipolaris oryzae (Breda de Haan) Shoemaker, the control of brown spot disease, the tillering, the number of panicles and the yield increase in rice were evaluated under laboratory and field conditions. In vitro, the growth of both fungi was completely inhibited by the EO of C. citrinus and C. citratus at 4,520 μg/ml and 452 μg/ml, respectively. For solvent extracts, the ETHE of C. citrinus was the most active and inhibited 80–85 % of the fungal growth followed by the CWE of C. citratus with 77 % and 36 % diameter reduction against B. oryzae and A. padwickii, respectively at 10 000 μg/ml. Under laboratory conditions, seed treatment with the EO of C. citrinus reduced the incidence of B. oryzae in seeds by 85–100 % compared to the non-treated controls. Similarly, the seed treatment increased the germination of an irrigated rice cultivar by 10.6 %, whereas the percentage of germinated seeds of upland rice was not significantly affected. The highest germination (85–94 %) was found in the non-treated and treated samples with a low incidence (0–4 %) of B. oryzae. Under field conditions, the combined use of the essential oil of C. citrinus as a seed treatment and spraying the plants with 2 % ethanol followed by 2 % (w/v) aqueous extracts of C. citrinus or C. citratus increased the emergence, tillering, panicles/plant and the grain yield by 25–55 % of the irrigated rice. In addition, the brown spot severity was reduced by 36–42 %. For the upland rice, the treatments led to similar results with the grain yield increase of 54–137 % and 20–80 % reduction in the brown spot severity. From our results, we concluded that the EO and solvent extracts of C. citrinus and C. citratus have potential as control agents against brown spot and other seed-borne fungal diseases in rice under both conventional and organic farming.

Five rice cultivars, one hybrid (WR96), three modern (BR16, BR26, and BRRI Dhan27) and one local (Pari) were screened for reaction to brown leaf spot disease caused by Cochliobolus miyabeanus and performance of yield-related characters. The severity of brown leaf spot varied with growth stages of rice plant as well as different cultivars tested under field condition. Low disease severity was observed at maximum tillering stage compared to moderate to high at dough stage, with hybrid cultivar WR96 showing highest disease, while local cultivar Pari had the lowest. Brown spot disease severity in different cultivars under induced epiphytic condition also followed the similar trend. The results also revealed that most of the yield-contributing characters examined showed wide variations among the cultivars. Modern cultivar BR16 produced the highest panicle length, number of grain per panicle and grain yield per hectare. At the same time as local cultivar Pari generated the lowest number of tiller per plant, pan...

α-1,3-Glucan, a component of the fungal cell wall, is a refractory polysaccharide for most plants. Previously, we showed that various fungal plant pathogens masked their cell wall surfaces with α-1,3-glucan to evade plant immunity. This surface accumulation of α-1,3-glucan was infection specific, suggesting that plant factors might induce its production in fungi. Through immunofluorescence observations of fungal cell walls, we found that carrot (Daucus carota) extract induced the accumulation of α-1,3-glucan on germlings in Colletotrichum fioriniae, a polyphagous fungal pathogen that causes anthracnose disease in various dicot plants. Bioassay-guided fractionation of carrot leaf extract successfully identified two active substances that caused α-1,3-glucan accumulation in this fungus: lutein, a carotenoid widely distributed in plants, and stigmasterol, a plant-specific membrane component. Lutein, which had a greater effect on C. fioriniae, also induced α-1,3-glucan accumulation in other Colletotrichum species and in the phylogenetically distant rice pathogen Cochliobolus miyabeanus, but not in the rice pathogen Magnaporthe oryzae belonging to the same phylogenetic subclass as Colletotrichum. Our results suggested that fungal plant pathogens reorganize their cell wall components in response to specific plant-derived compounds, which these pathogens may encounter during infection.

International audience; The fungus Cochliobolus miyabeanus causes severe leaf spot disease on rice (Oryza sativa) and two North American specialty crops, American wildrice (Zizania palustris) and switchgrass (Panicum virgatum). Despite the importance of C. miyabeanus as a disease-causing agent in wildrice, little is known about either the mechanisms of pathogenicity or host defense responses. To start bridging these gaps, the genome of C. miyabeanus strain TG12bL2 was shotgun sequenced using Illumina technology. The genome assembly consists of 31.79 Mbp in 2,378 scaffolds with an N50 = 74,921. It contains 11,000 predicted genes of which 94.5% were annotated. Approximately 10% of total gene number is expected to be secreted. The C. miyabeanus genome is rich in carbohydrate active enzymes, and harbors 187 small secreted peptides (SSPs) and some fungal effector homologs. Detoxification systems were represented by a variety of enzymes that could offer protection against plant defense compounds. The non-ribosomal peptide synthetases and polyketide synthases (PKS) present were common to other Cochliobolus species. Additionally, the fungal transcriptome was analyzed at 48 hours after inoculation in planta. A total of 10,674 genes were found to be expressed, some of which are known to be involved in pathogenicity or response to host defenses including hydrophobins, cutinase, cell wall degrading enzymes, enzymes related to reactive oxygen species scavenging, PKS, detoxification systems, SSPs, and a known fungal effector. This work will facilitate future research on C. miyabeanus pathogen-associated molecular patterns and effectors, and in the identification of their corresponding wildrice defense mechanisms.

The plant hormone abscisic acid (ABA) is involved in an array of plant processes, including the regulation of gene expression during adaptive responses to various environmental cues. Apart from its well-established role in abiotic stress adaptation, emerging evidence indicates that ABA is also prominently involved in the regulation and integration of pathogen defense responses. Here, we demonstrate that exogenously administered ABA enhances basal resistance of rice (Oryza sativa) against the brown spot-causing ascomycete Cochliobolus miyabeanus. Microscopic analysis of early infection events in control and ABA-treated plants revealed that this ABA-inducible resistance (ABA-IR) is based on restriction of fungal progression in the mesophyll. We also show that ABA-IR does not rely on boosted expression of salicylic acid-, jasmonic acid -, or callose-dependent resistance mechanisms but, instead, requires a functional Gα-protein. In addition, several lines of evidence are presented suggesting that ABA steers its positive effect on brown spot resistance through antagonistic cross talk with the ethylene (ET) response pathway. Exogenous ethephon application enhances susceptibility, whereas genetic disruption of ET signaling renders plants less vulnerable to C. miyabeanus attack, thereby inducing a level of resistance similar to that observed on ABA-treated wild-type plants. Moreover, ABA treatment alleviates C. miyabeanus-induced activation of the ET reporter gene EBP89, while derepression of pathogen-triggered EBP89 transcription via RNA interference-mediated knockdown of OsMPK5, an ABA-primed mitogen-activated protein kinase gene, compromises ABA-IR. Collectively, these data favor a model whereby exogenous ABA enhances resistance against C. miyabeanus at least in part by suppressing pathogen-induced ET action in an OsMPK5-dependent manner.

Thirteen endophytic non sporulating fungi present in healthy seeds of rice cultivars, popular in coastal India, were isolated by ‘host tissue transplant ’method. Those were identified by the molecular technique and phylogenetic analysis by Phylogeny fr software was done. The phylogeny tree of 32 taxa including 27 Dendryphiella species and five Cochliobolus miyabeanus spp. was rendered. There were two major groups. Five Cochliobolus miyabeanus spp. (out group) were together the smaller cluster whereas all the Dendryphiella species with same internal node. The Dendryphiella group was further subdivided in two sub groups where 10 marine Dendryphiella and 17 terrestrial endophytic Dendryphiella grouped in two different sister groups. The high bootstrap values for each subcluster justified their grouping in the proper bunch. This analysis indicated that marine and ‘terrestrial endophytic Dendryphiella ’had common ancestors.

In order to study a relationship between soil nutrients and rice brown spot occurrence, paddy field soils, rice grains and straws collected from different paddy fields with different disease degrees of brown spots were analyzed for inorganic nutrients. Brown spot was prevalent in the rice grown in nutrient-deficient soils, which is especially low in macronutrient elements (phosphoric acid, potassium, silicic acids) and micronurients (calcium, magnesium). The soil, however, was high in sodium while organic nutrients and pH level were similar to others. The rice straws with severe brown spot were low in inorganics such as ferrous, copper, T-N, and while the rice grains with brown spot were low in ferrous, MgO, Zn, and Mn. In the analysis of field type and nitrogen level, the highest disease severity was found in sandy-type field soil, followed by salty-type field soil and disease severity decreased as application level of nitrogen fertilizer increased. As a summary, the most important factor for effective brown spot control in rice is maintenance of proper nutrients in sandy-type field and control of sodium level in salty-type field soil.

Fifty plant extracts, four oil cakes and eight antagonistic organisms were tested against Bipolaris oryzae (Cochliobolus miyabeanus), the causal agent of brown spot disease of rice. In vitro studies indicated that two leaf extracts, Nerium oleander and Pithecolobium dulce exerted the higher percent inhibition to mycelial growth (77.4, 75.1%) and spore germination (80.3, 80.0%) of B. oryzae. Among the four oil cake extracts tested in vitro against B. oryzae, neem cake extract showed the maximum inhibition percent to mycelial growth (80.18%) and spore germination (81.13%) of the pathogen followed by mahua cake extract, castor and gingelly cake extract. Trichoderma viride (Tv2) was significantly effective in inhibiting the mycelial growth (62.92%) and spore germination (77.03%) of the pathogen followed by Trichoderma harzianum (Th5) and Trichoderma reesei (Tr3). The promising leaf extracts, oil cake extracts and antagonistic microorganisms were further evaluated for their efficacies in disease management under glasshouse and field conditions. In glasshouse studies, post-infectional spraying of rice plants with neem cake extract, N. oleander leaf extract and T. viride (Tv2) was significantly effective in reducing the incidence of brown spot of rice by 66, 52 and 45 percent respectively. Two rounds of spraying of rice plants with neem cake extract, N. oleander leaf extract and T. viride (Tv2) in the field at initial appearance of disease and 15 days later reduced the incidence of brown spot (70, 53 and 48% disease reduction respectively) and increased the yield by 23, 18 and 15 percent respectively.

Ahn, I.-P., and Suh, S.-C. 2007. Calcium restores prepenetration morphogenesis abolished by methylglyoxal-bis-guanyl hydrazone in Cochliobolus miyabeanus infecting rice. Phytopathology 97:331-337. Cochliobolus miyabeanus forms a specialized infection structure, an appressorium, to infect its host rice plants. Curtailment of prepenetration development by spermidine and spermine was more evident in appressorium development and germination remained unaffected, whereas putrescine and methylglyoxal-bis-guanyl hydrazone (MGBG) impaired both morphogenetic events. Exogenous calcium nullified the inhibitory effect of MGBG on the prepenetration development in vitro and in vivo and the disease progression. High levels of polyamines were detected in freshly collected conidia, but the amounts were reduced during germination and appressorium formation. MGBG fortified the decrease of polyamines within conidia under development and calcium amendment did not affect the reduction. Hard-surface contact augmented messenger RNA synthesis of calmodulin gene (CmCaM) and protein kinase C (PKC) activity in germinating or appressorium-forming conidia. Calcium restored transcription of CmCaM and upregulation of PKC activity suppressed by MGBG. Taken together, fine-tuning of intracellular polyamine transition is indispensable for the conidial germination and appressorium formation in C. miyabeanus. Biochemical and molecular analyses revealed that the MGBG-acting site or sites are upstream of Ca2+-dependent signaling pathways regulating prepenetration morphogenesis of C. miyabeanus causing rice brown leaf spot. Additional keywords: Bipolaris oryzae, Oryza sativa.

Mepanipyrim exhibited excellent disease control activity against Botrytis cinerea, but poor activity against Cochliobolus miyabeanus; however, the mycelial growth of C. miyabeanus was inhibited more strongly than that of B. cinerea. Therefore, disease control efficacy by mepanipyrim in vivo is not correlated with mycelial growth inhibition in vitro. While mepanipyrim prevented pectinase secretion in B. cinerea at 0.1–1 μg/ml, it did not interfere with secretion in C. miyabeanus, even at 100 μg/ml, indicating that its action is an important mechanism in disease control. Mepanipyrim affected the uptake of glucose and phenylalanine in the mycelia of both pathogens at higher doses. Thus, a secondary action of mepanipyrim may bring about mycelial growth inhibition in vitro.

Bacillus vallismortis EXTN-1, a biocontrol agent in cucumber, tomato and potato was tested in rice pathosystem against rice fungal pathogens viz. Magnaporthe grisea, Rhizoctonia solani and Cochliobolus miyabeanus. Apart from increasing the yield in the bacterized plants (11.6-12.6% over control), the study showed that EXTN- 1 is effective in bringing about disease suppression against all the tested fungal pathogens. EXTN-1 treatment resulted in 52.11% reduction in rice blast, 83.02% reduction in sheath blight and 11.54% decrease in brown spot symptoms. As the strain is proven as an inducer for systemic resistance based on PR gene expression in Arabidopsis and tobacco models, it is supposed that a similar mechanism works in rice, bringing about disease suppression. The strain could be used as a potent biocontrol and growth-promoting agent in rice cropping system.

Responses of rice to Magnaporthe grisea and Cochliobolus miyabeanus were compared. In Tetep, a rice cultivar resistant to both fungi, pathogen inoculation rapidly triggered the hypersensitive reaction (HR), resulting in microscopic cell death. In rice cv. Nakdong, susceptible to both pathogens, M. grisea did not cause HR, whereas C. miyabeanus caused rapid cell death similar to that associated with HR, which appeared similar to that observed in cv. Tetep, yet failed to block fungal ramification. Treatment with conidial germination fluid (CGF) from C. miyabeanus induced rapid cell death in both cultivars, suggesting the presence of phytotoxins in CGF. Pretreatment of cv. Nakdong with CGF significantly increased resistance to M. grisea, while the same treatment was ineffective against C. miyabeanus. Similarly, in cv. Nakdong, benzothiadiazole (BTH) significantly increased resistance to M. grisea, but was ineffective against C. miyabeanus. Methyl jasmonate (MeJA) treatment appeared to be ineffective against either fungus. Increased resistance of cv. Nakdong to M. grisea by BTH or CCF treatment was correlated with more rapid induction of three monitored PR genes. Application of MeJA resulted in the expression of JAmyb in cv. Nakdong being induced faster than in untreated plants in response to M. grisea infection. In contrast, the expression pattern of the PR and JAmyb genes in response to C. miyabeanus was nearly identical between cvs. Nakdong and Tetep, and neither BTH nor MeJA treatment significantly modified their expression patterns in response to C. miyabeanus infection. Our results suggest that rice employs distinct mechanisms for its defense against M. grisea versus C. miyabeanus.

The antifungal activity of 30 medicinal plants belonging to different families was tested in vitro on the phytopathogenic fungus Rhizoctonia solani. The results revealed that protein extract from the galls of Quercus infectoria belonging to Fagaceae family was highly effective in inhibiting the mycelial growth of R. solani. The gall extract of Q. infectoria also inhibited the growth of other agronomically important fungal pathogens viz. Fusarium oxysporum, Cochliobolus miyabeanus, Macrophomina phaseolina, Colletotrichum gloeosporioides, Magnaporthe salvinii, Cochliobolus lunatus, Alternaria solani, Pythium aphanidermatum and Colletotrichum falcatum. A 29-kDa glycoprotein was purified from the galls of Q. infectoria by ammonium sulphate fractionation followed by gel filtration on Sephadex G-50 column. The purified protein showed the absorption maxima at 640 nm and 308 nm. The purified protein was stable even after heating at 100 °C for 10 min or autoclaving at 121 °C for 20 min. When the 29-kDa protein was treated with NaIO4 and pronase the antifungal activity was drastically reduced. The 29-kDa protein inhibited the mycelial growth of R. solani and C. miyabeanus at 2 µg level.

Switch grass (Panicum virgatum) is a native, perennial warm-season grass used for hay, summer grazing, soil conservation, and wildlife habitat. It is also being developed as a biomass crop for renewable energy. A previously unreported leaf-spot disease on switch grass was observed in North Dakota plantings. From 1999 to 2002, a fungus was consistently isolated from leaf spots on switch grass. It was identified as Bipolaris oryzae (teleomorph: Cochliobolus miyabeanus). Conidial morphology agreed with that of B. oryzae, and comparison of DNA sequences from glyceraldehyde-3-phosphate dehydrogenase also indicated that the isolates were B. oryzae. Greenhouse pathogenicity tests confirmed the ability of B. oryzae to cause symptoms of leaf spot on switch grass. This is the first time that B. oryzae, a common pathogen on rice (Oryza sativa), is reported to cause a disease on switch grass. This pathogen has the potential to diminish yields of switch grass under intensive plantings for high biomass production, part...

This study was carried out to investigate factors affecting disease development of rice brown spot caused by Cochliobolus miyabeanus in the field and under the greenhouse conditions. In normal paddy fields, the pathogen was more frequently isolated from dried rice straws than wet straws after overwintering. The pathogen was also readily isolated from wet rice straws of salty paddy fields although infection rate of the fields was low. The disease symptom was first observed at 2 to 3days after inoculation when the seedlings were kept for 6 hours in dew chamber. The number of lesions peaked at 14 hr incubation in cultivars Chucheongbyeo and 24 hr incubation in cultivar Daesanbyeo and Tetep. Sixty three-d-old seedlings were more susceptible than 21 to 49-d-old seedlings regardless of cultivar. There was some difference in number of lesions observed, but the lesions were evenly spread regardless of leaf position. Light on the rice seedlings at 3,000 lux or 6,000 lux showed more severe symptoms than those in the dark.

The oil extract from seeds of Azadirachta indica as well as water and ethanol leaf extracts of the plant were effective in reducing the radial growth of Cochliobolus miyabeanus in culture and in checking the spread of brown spot disease in rice. The oil extract was found to be the most effective, followed by ethanol leaf extract, in inhibiting the growth of the pathogen in vitro and in checking the development of the disease in vivo . Although the cold water extract from the plant leaf reduced the radial growth of the pathogen in culture, it failed to reduce the incidence and severity of the disease as effectively as the oil and ethanol extracts. The oil and ethanol extracts compared favourably with carbendazim (Bavistin) at 0.1% a.i and have the potential for control of brown spot disease of rice in the field.

Summary Although numerous studies have shown the ability of silicon (Si) to mitigate a wide variety of abiotic and biotic stresses, relatively little is known about the underlying mechanism(s). Here, we have investigated the role of hormone defense pathways in Si-induced resistance to the rice brown spot fungus Cochliobolus miyabeanus. To delineate the involvement of multiple hormone pathways, a multidisciplinary approach was pursued, combining exogenous hormone applications, pharmacological inhibitor experiments, time-resolved hormone measurements, and bioassays with hormone-deficient and/or -insensitive mutant lines. Contrary to other types of induced resistance, we found Si-induced brown spot resistance to function independently of the classic immune hormones salicylic acid and jasmonic acid. Our data also rule out a major role of the abscisic acid (ABA) and cytokinin pathways, but suggest that Si mounts resistance to C. miyabeanus by preventing the fungus from hijacking the rice ethylene (ET) machinery. Interestingly, rather than suppressing rice ET signaling per se, Si probably interferes with the production and/or action of fungal ET. Together our findings favor a scenario whereby Si induces brown spot resistance by disarming fungal ET and argue that impairment of pathogen virulence factors is a core resistance mechanism underpinning Si-induced plant immunity.

Necrotrophic fungal pathogen Cochliobolus miyabeanus causes brown spot disease in rice leaves upon infection, resulting in critical rice yield loss. To better understand the rice-C. miyabeanus interaction, we employed proteomic approaches to establish differential proteomes of total and secreted proteins from the inoculated leaves. The 2DE approach after PEG-fractionation of total proteins coupled with MS (MALDI-TOF/TOF and nESI-LC-MS/MS) analyses led to identification of 49 unique proteins out of 63 differential spots. SDS-PAGE in combination with nESI-LC-MS/MS shotgun approach was applied to identify secreted proteins in the leaf apoplast upon infection and resulted in cataloging of 501 unique proteins, of which 470 and 31 proteins were secreted from rice and C. miyabeanus, respectively. Proteins mapped onto metabolic pathways implied their reprogramming upon infection. The enzymes involved in Calvin cycle and glycolysis decreased in their protein abundance, whereas enzymes in the TCA cycle, amino acids, and ethylene biosynthesis increased. Differential proteomes also generated distribution of identified proteins in the intracellular and extracellular spaces, providing a better insight into defense responses of proteins in rice against C. miyabeanus. Established proteome of the rice-C. miyabeanus interaction serves not only as a good resource for the scientific community but also highlights its significance from biological aspects.

International audience; Much has been written and said of the importance of crop management to sustain plant health, a fundamental for feeding a world population expected to reach around nine billion by 2050. Yet, the two elements, crop management and plant health management, are generally addressed as two distinct entities, the former often being seen as a source for options for the latter. Agriculture of the Middle Age (900-1600 CE) seems not always to have distinguished crop and plant health management, considering them instead as a whole. In this article, these elements are addressed as parts of production situations, which represent a much broader level of integration for systems analysis. At the production situation level, plant disease epidemics, or their suppression, can both be seen as consequences of crop management. Modern agriculture tends to distinguish specific plant health management techniques (which are often over-emphasized) and general, non-specific ones (which are often downplayed). The former (e.g., specific host plant resistances, biological control, pesticides) may be overcome rather rapidly by plant pathogens, while the latter (e.g., crop rotation, tillage), with often progressive, incomplete, and confounded effects, may constitute the basis of sustainable management of plant health. A simulation modelling framework, with two model structures, is used to highlight the three components of disease management outlined by Berger (Annual Review of Phytopathology, 15, 165-183, 1977): (1) eliminate or reduce the initial inoculum or delay its appearance, (2) slow the rate of disease increase, and (3) shorten the time of exposure of the crop to the pathogen. The degree of involvement of these three components is further discussed in three groups of examples in plant production systems that significantly contribute to global food provisioning and food security. A few modelling outputs and evaluation of these examples suggest that, even in the most specialized production situations, both specific and non-specific plant health management strategies are necessary for sustained health of our crops and for durable performances of agricultural systems.

A gamma-ray-induced rice mutant, M1009, which spontaneously forms brown specks on leaves was isolated. In appearance, these lesions very much resemble those that are produced during the hypersensitive resistance response of rice to pathogens. A similar phenotype has been associated with a number of mutants in maize or Arabidopsis, which have been briefly categorized as disease lesion mimics. Brown speck formation was suppressed at temperatures of 25°C and above, and was severe at 20°C. Light irradiation is also required to initiate brown specks. In addition, the mutant exhibits heightened resistance to two major fungal pathogens of rice, Magnaporthe grisea and Cochliobolus miyabeanus. Genetic characterization of the mutant has indicated that its les-bs (lesion-brown speck) phenotype segregates as a monogenic recessive mutation.

The red pigmented strain F-1-1 of Serratia marcescens shows a strong antagonism against Phytophthora capsici, the pathogen of damping-off of cucumber seedlings, and its antagonistic activity is due to the production of antibiotic red pigments. In the present study, purification and identification of the red pigments produced by the strain F-1-1 were performed. The water-insoluble red pigments were extracted from cultures on agar plates with ethanol and then chloroform, and purified by silica gel column chromatography. An active substance was isolated using the antifungal activity assay and was identified as prodigiosin, a known compound based on the spectral data. Purified prodigiosin inhibited the germination of cystospores and the growth of the fungal hyphae of P. capsici. It also showed antimicrobial activity against some plant-pathogenic fungi including Cochliobolus miyabeanus, Pythium spinosum, P. ultimum and against plant pathogenic bacteria including, Clavibacter michiganensis subsp. michiganensis and Erwinia carotovora subsp. carotovora. These results suggest that S. marcescens F-1-1 could be used as a biocontrol agent against many plant diseases other than cucumber damping-off.

Although silicon is not considered an essential element; plant development, growth and yield has been increased in many graminaceous and some nongraminaceous crop species. Silicon also is known to reduce plant diseases especially in rice. Silicon fertilization has become a routine practice in Florida rice production. The information within provides an overview on the history of silicon in Florida, application of silicon and disease suppression by silicon and its interaction with fungicides and rice genotypes. Although the focus is on rice and organic soils, this information should be of interest to those working on other grass crops on organic soils as well as rice production on weathered, low-silicon mineral soils. An outlook and future research needs also are presented.

Epicoccum sp. showed in vitro antagonism to the rice pathogens Magnaporthe oryzae, Rhizoctonia solani, Sarocladium oryzae, Monographella albescens and Cochliobolus miyabeanus in dual culture. The colony growth of the rice blast fungus, M. oryzae, was reduced by 42.50%. The lethal doses (LD50) determined based on probit-log dosage response curves at 3 and 6 hours after germination were 393.0 and 326.6 ppm, respectively. The crude extract (600 ppm) reduced appressorial formation by 95.68%. A greenhouse experiment comparing the relative efficiency of conidial suspension and crude extract of Epicoccum sp. in the suppression of leaf blast showed no statistical difference between both application methods. However, the crude extract of Epicoccum sp. (4000 ppm) 48 hours before the application of M. oryzae induced resistance and suppressed leaf blast by 97.6%. Scanning electron microscopy of rice leaves inoculated with crude extract of Epicoccum sp. and challenged with M. oryzae showed appressorial deformation on penetration phase. Peroxidase and β-1,3-glucanase activities increased in plants sprayed with crude extract of Epicoccum sp., 24 hours after the application of the challenger. PAL as well as chitinase activities increased 72 hours after challenge inoculation. Epicoccum sp. was shown to be a potential antagonist and inducer of resistance against M. oryzae.

Rice has low productivity in Tocantins due to the low genetic potential of the varieties and high susceptibility to diseases. Therefore, the objective of this study was to evaluate a rice nuclear collection of highlands in relation to its agronomic and resistance characters to the main diseases in the south of Tocantins State. It was used the randomized blocks design with ninety treatments (genotypes) and three repetitions under highlands conditions in southern Tocantins. The following agronomic characters were evaluated: average flowering, height and productivity of the plants. The evaluated diseases were: leaf and panicle blast, brown spot, leaf scald and grain spots. The Scott-Knott test showed high variability in productivity and in resistance to the diseases. The Tocher Method grouped the 90 genotypes, according to the diseases and productivity, in 10 different groups. These results indicate the existence of significant genetic variability for these characters in the genotypes.Read Complete Article at ijSciences: V2201303128

Wall-associated kinases comprise a sub-family of receptor-like kinases that function in plant growth and stress responses. Previous studies have shown that the rice wall-associated kinase, OsWAK25, interacts with a diverse set of proteins associated with both biotic and abiotic stress responses. Here, we show that wounding and BTH treatments induce OsWAK25 transcript expression in rice. We generated OsWAK25 overexpression lines and show that these lines exhibit a lesion mimic phenotype and enhanced expression of rice NH1 (NPR1 homolog 1), OsPAL2, PBZ1 and PR10. Furthermore, these lines show resistance to the hemibiotrophic pathogens, Xanthomonas oryzae pv. oryzae (Xoo) and Magnaporthe oryzae, yet display increased susceptibility to necrotrophic fungal pathogens, Rhizoctonia solani and Cochliobolus miyabeanus.

Rice brown leaf spot is a major disease in the rice paddy field. The causal agent Cochliobolus miyabeanus is an ascomycete fungus and a representative necrotrophic pathogen in the investigation of rice-microbe interactions. The aims of this research were to identify a quantitative evaluation method to determine the amount of C. miyabeanus proliferation in planta and determine the method's sensitivity. Real-time polymerase chain reaction (PCR) was employed in combination with the primer pair and Taqman probe specific to CmSCD1, a C. miyabeanus unigene encoding SCYTALONE DEHYDRATASE, which is involved in fungal melanin biosynthesis. Comparative analysis of the nucleotide sequences of CmSCD1 from Korean strains with those from the Japanese and Taiwanese strains revealed some sequence differences. Based on the crossing point (CP) values from Taqman real-time PCR containing a series of increasing concentrations of cloned amplicon or fungal genomic DNA, linear regressions with a high level of reliability (R(2)0.997) were constructed. This system was able to estimate fungal genomic DNA at the picogram level. The reliability of this equation was further confirmed using DNA samples from both resistant and susceptible cultivars infected with C. miyabeanus. In summary, our quantitative system is a powerful alternative in brown leaf spot forecasting and in the consistent evaluation of disease progression.

Fungi, yeast, and bacteria were cultured from field-collected, discolored (pecky) rice, Oryza sativa L., kernels and from stylets and saliva of the rice stink bug, Oebalus pugnax (F.). A sterilized chrome wire was used to wound and introduce selected inocula into developing kernels on rice panicles. Typical kernel discolorations could not be induced by inoculation with bacterial or yeast isolates. When the caryopsis was wounded during inoculation, several fungi caused discoloration comparable to that of pecky kernels collected from rice fields. Fusarium oxysporum Schlect, caused the greatest discoloration followed by Cochliobolus miyabeanus (Ito & Kuribayashi) Drechsler ex Dastur, Curvularia lunata (Wakker) Boedijn, Alternaria alternate (Fr.) Keissler, and A. Padwickii (Ganguly) M. B. Ellis. Symptoms typifying classical fungal discoloration peaked 5 to 10 dafter anthesis, and classical insect damage symptoms peaked 15 dafter anthesis when rice stink bugs were caged on plants 0, 5, 10, or 15 dafter anthesis. In field tests, density of rice stink bug infestations was associated positively with incidence of discolorations historically attributed to insects and fungi. These data indicate that discolorations in pecky rice result from fungi being introduced at the time of feeding of O. pugnax by means of a loose vector relationship.

The ecological behaviour of Cochliobolus miyabeanus (Ito et Kurib.) Drechsl. ex Dast. (Bipolaris oryzae (Breda de Haan) Shoem.), causal agent of brown spot of rice (Oryza sativa L.), was investigated in a tidal mangrove swamp at Rokupr Experimental Farm, northwestern Sierra Leone. Disease increased where transplanting was delayed. This was probably due to the coincidence of flowering with favourable environmental conditions for disease development from November to February. Nitrogen fertilization at 40kg N ha−1 significantly (P = 0.05) reduced C. miyabeanus incidence in 1983, but not in 1984 and 1985 cropping seasons at the same site. Grain yields of ROK 5, a medium‐duration improved rice cultivar (ca. 150 days) were significantly reduced in late‐transplanted rice crops (September to November) in spite of adequate N fertilization. Mangrove mud was not an important source of C. miyabeanus propagules. Incidence of leaf scald caused by Monographella albescens (Thum) Parkinson, Sivanesan and Booth (M...

We produced a polyclonal antibody which reacts with antigens of mycelia of the rice blast fungus. Using this antibody, we developed an ELISA method which can measure small amounts of mycelia of Magnaporthe grisea (Hebert) Barr in a rice leaf. More than 0.1μg mycelia of the rice blast fungus in 100mg of fresh leaf can be measured by this method. The antibody, acquired using lyophilized mycelia of the fungus, reacts as well with fresh mycelia, conidia, and even those of different races as it does with lyophilized ones. But it does not react with mycelia of several fungal pathogens of the rice plant. Thanatephorus cucumeris (Frank) Donk, Rhizoctonia sp., Cochliobolus miyabeanus (Ito et Kuribayashi) Drechsler ex Dastur, Sphaerulina oryzina Hara and Nigrospora oryzae (Berkeley et Broome) Petch do not react with the antibody at 20μg dry weight mycelia/ml of PBS (pH 7.5) containing 0.05% Tween 80. But at about 2mg/ml, the antibody slightly reacts with mycelia of Sphaerulina oryzina and Nigrospora oryzae. This method should be useful in understanding the interaction between blast fungus and rice plant.

The application of bark-compost to farm land has been reported to suppress soil-borne diseases, including Fusarium wilts. The “bacterial type” microflora of the compost, that includes many antagonists, has been considered to play the major role in suppression of soil-borne plant pathogens. To elucidate the suppressive effect of hemlock bark-compost on soil-borne pathogenic fungi, one of the antifungal agents in bark-compost was isolated by preparative HPLC. From 2D-nuclear magnetic resonance spectra and high resolution mass spectrometry, the antifungal agent was determined to be 24R-ethylcholest-5-en-3β-ol (silosterol). Sitosterol is a normal secondary plant metabolite found throughout the plant kingdom. Furthermore, using reverse-phase HPLC, we found considerable amounts of cholesterol, campesterol and stigmasterol in acetone extracts of the bark-compost. These typical sterols had strong in vitro antifungal activity against Fusarium oxysporum f. sp. cucumerinum . We investigated the antimicrobial spectrum of cholesterol. It was active against Cochliobolus miyabeanus, Gibberella zeae, Glomerella cingulata, Alternaria alternata and Rhizoctonia solani , fungi of interest to plant and human pathologists. It was inactive against yeasts and procaryotic organisms, based on a disk-diffusion technique. We propose that there might be a parallel between this selective antimicrobial activity of the sterol and the formation of a “bacterial type” microflora of the compost. These findings suggest that the typical sterols, such as cholesterol, may function as defensive substances against pathogenic fungi at the external surface of higher eucaryotes, including the cuticle of higher plants and in kelatinized squames of the mammalian epidermis.

The rice brown spot disease caused by Cochliobolus miyabeanus (Ito et Kurib.) Drechsl. ex Dast. = (Helminthosporium oryzae Breda da Haan) is a key disease in mangrove swamp rice ecosystem in Sierra Leone. On average, grain yield losses of 8.2–23.0% were caused by the pathogen in mangrove swamps of northwestern Sierra Leone in the 1983, 1984 and 1985 cropping seasons. Disease attack was closely linked with adverse soil conditions and favourable meteorological factors of long dew periods and moderate temperatures. However, attack by C. miyabeanus at the reproductive growth stage of rice caused the most damage because of direct effect on the grains.

Glufosinate ammonium diminished developments of rice (Oryza sativa) blast and brown leaf spot in 35S:bar-transgenic rice. Pre- and postinoculation treatments of this herbicide reduced disease development. Glufosinate ammonium specifically impeded appressorium formation of the pathogens Magnaporthe grisea and Cochliobolus miyabeanus on hydrophobic surface and on transgenic rice. In contrast, conidial germination remained unaffected. Glufosinate ammonium diminished mycelial growth of two pathogens; however, this inhibitory effect was attenuated in malnutrition conditions. Glufosinate ammonium caused slight chlorosis and diminished chlorophyll content; however, these alterations were almost completely restored in transgenic rice within 7 d. Glufosinate ammonium triggered transcriptions of PATHOGENESIS-RELATED (PR) genes and hydrogen peroxide accumulation in transgenic rice and PR1 transcription in Arabidopsis (Arabidopsis thaliana) wild-type ecotype Columbia harboring 35S:bar construct. All transgenic Arabidopsis showed robust hydrogen peroxide accumulation by glufosinate ammonium. This herbicide also induced PR1 transcription in etr1 and jar1 expressing bar; however, no expression was observed in NahG and npr1. Fungal infection did not alter transcriptions of PR genes and hydrogen peroxide accumulation induced by glufosinate ammonium. Infiltration of glufosinate ammonium did not affect appressorium formation of M. grisea in vivo but inhibited blast disease development. Hydrogen peroxide scavengers nullified blast protection and transcriptions of PR genes by glufosinate ammonium; however, they did not affect brown leaf spot progression. In sum, both direct inhibition of pathogen infection and activation of defense systems were responsible for disease protection in bar-transgenic rice.

Three isolates of Pseudomonas aeruginosa were used for seed treatment of rice; all showed plant growth promoting activity and induced systemic resistance in rice against Rhizoctonia solani G5 and increased seed yield. Production of salicylic acid (Sal) by P. aeruginosa both in vitro and in vivo was quantified with high performance liquid chromatography. All three isolates produced more Sal in King's B broth than in induced roots. Using a split root system, more Sal accumulated in root tissues of bacterized site than in distant roots on the opposite site of the root system after 1 d, but this difference decreased after 3 d. Sal concentration 0-200 g/L showed no inhibition of mycelial growth of R. solani in vitro, while ator =300 g/L it inhibited it. P. aeruginosa-pretreated rice plants challenged inoculation with R. solani (as pathogen), an additional increase in the accumulation of peroxidase was observed. Three pathogenesis-related peroxidases in induced rice plants were detected; molar mass of these purified peroxidases was 28, 36 and 47 kDa. Purified peroxidase showed antifungal activity against phytopathogenic fungi R. solani, Pyricularia oryzae and Helminthosporium oryzae.

NPS6, encoding a nonribosomal peptide synthetase, is a virulence determinant in the maize (Zea mays) pathogen Cochliobolus heterostrophus and is involved in tolerance to H2O2. Deletion of NPS6 orthologs in the rice (Oryza sativa) pathogen, Cochliobolus miyabeanus, the wheat (Triticum aestivum) pathogen, Fusarium graminearum, and the Arabidopsis thaliana pathogen, Alternaria brassicicola, resulted in reduced virulence and hypersensitivity to H2O2. Introduction of the NPS6 ortholog from the saprobe Neurospora crassa to the Δnps6 strain of C. heterostrophus restored wild-type virulence to maize and tolerance to H2O2, demonstrating functional conservation in filamentous ascomycete phytopathogens and saprobes. Increased sensitivity to iron depletion was identified as a conserved phenotype of Δnps6 strains. Exogenous application of iron enhanced the virulence of Δnps6 strains of C. heterostrophus, C. miyabeanus, F. graminearum, and A. brassicicola to each host. NPS6 is responsible for the biosynthesis of extracellular siderophores by C. heterostrophus, F. graminearum, and A. brassicicola. Application of the extracellular siderophore of A. brassicicola restored wild-type virulence of the ΔAbnps6 strain to Arabidopsis. It is proposed that the role of extracellular siderophores in fungal virulence to plants is to supply an essential nutrient, iron, to their producers in planta and not to act as phytotoxins, depriving their hosts of iron.