Rice blast (Magnaporthe oryzae) can cause large losses in crop yields, especially in upland rice systems. Avirulent strains of M. oryzae can induce resistance to subsequent attacks by virulent ...strains in plants. This study aimed to investigate the defense responses in upland rice challenged with a virulent strain of M. oryzae after acclimation with an avirulent strain. The avirulent strain decreased rice blast severity in the challenged plants. Induced resistance was characterized by a hypersensitive response and early accumulation of phenolic compounds. Scanning electron microscopy showed that M. oryzae conidia germinate and form appressoria, but do not colonize leaf tissues. The activities of pathogenesis-related proteins, total phenolic compounds, and salicylic acid (SA) were affected by acclimation to the avirulent strain. The activities of β-1,3-glucanase, phenylalanine ammonia-lyase, and peroxidase, as well as the SA levels explained most of the variability in the rice plant responses to M. oryzae. In addition, OsXa13, OsMAPKKK74, OsAOS2, OsACO7, and OsMAS1 expression was modulated depending on the virulence of the M. oryzae strains. This modulation in gene expression is critical for infection and some of these mechanisms are targeted by effectors, resulting in enhanced susceptibility and pathogen infection. These results have practical importance in plant-pathogen interaction studies to identify resistance-relevant mechanisms against M. oryzae in upland rice.
Abstract Purple ipe (Handroanthus impetiginosus) is an important tree species in Cerrado biome conservation and very popular at the landscaping and urban afforestation. However, its micropropagation ...is affected by pathogens, such as Oidium sp. The aim this study was evaluate the efficiency of seed treatments in the control of powdery mildew of purple ipe obtained by micropropagation. The symptoms were observed during in vitro germination, a Koch’s postulates were performed for confirm the pathogenicity, colonization of the pathogen on the leaves was analyzed in optical and scanning microscopes and a scale to evaluate severity was proposed. Two experiments were realized to powdery mildew control using a completely randomized design, with 30 replicates. First experiment: Seeds were treated with ethanol (Et), chlorothalonil + thiophanate-methyl (C+TM), and sodium hypochlorite (NaOCl); second experiment: Seeds were treated with Et, NaOCl, C+TM, and neem oil. Disease severity and area under the disease progress curve (AUDPC) were assessed in both experiments. Disease symptoms and typical pathogen structures were observed, and the pathogenicity was confirmed. The disease severity was reduced by 30.78% in 1.5% neem oil for 10 min when compared with C+TM for 15 min. We conclude that neem oil can be a strategy sustainable for the control of powdery mildew in purple ipe in tissue culture.
Resumo O ipê-roxo, Handroanthus impetiginosus, é uma espécie arbórea importante na conservação do bioma Cerrado e muito procurada no paisagismo e arborização urbana. No entanto, sua micropropagação é afetada por patógenos, como Oidium sp. O objetivo deste estudo foi avaliar a eficácia dos tratamentos em sementes no controle do oídio do ipê roxo por micropropagação. Os sintomas foram observados durante a germinação in vitro, os postulados de Koch foram realizados para confirmação da patogenicidade, a colonização do patógeno nas folhas foi analisada em microscópios ópticos e de varredura e uma escala para avaliação de severidade foi proposta. Foram realizados dois ensaios para controle do oídio em delineamento inteiramente casualizado, com 30 repetições. Primeiro experimento: sementes foram tratadas com etanol, clorotalonil + tiofanato-metílico (C+TM) e hipoclorito de sódio; segundo experimento: sementes foram tratadas com etanol, NaOCl, C+TM e óleo de neem. A severidade da doença e a área abaixo da curva de progresso da doença foram quantificadas. Os sintomas da doença e estruturas típicas do patógeno foram observados e a patogenicidade foi confirmada. A severidade da doença foi reduzida em 30,78% com 1,5% de óleo de neem por 10 min quando comparado com C + TM por 15 min. Concluímos que o óleo de neem pode ser uma estratégia sustentável para o controle do oídio em ipê roxo na cultura de tecidos.
ABSTRACT 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.
Filamentous fungi exhibit unparalleled potential as cell factories for protein production, owing to their adeptness in protein secretion and remarkable proficiency in post-translational ...modifications. This review delineates the role of filamentous fungi in bio-input technology across different generations and explores their capacity to generate secondary metabolites. Our investigation highlights filamentous fungi as frontrunners in the production of bioactive compounds, emphasizing the imperative nature of elucidating their metabolic repertoire. Furthermore, we delve into common strategies for genetic transformation in filamentous fungi, elucidating the underlying principles, advantages, and drawbacks of each technique. Taking a forward-looking approach, we explore the prospects of genome engineering, particularly the CRISPR-Cas9 technique, as a means to propel protein secretion in filamentous fungi. Detailed examination of the protein secretion pathways in these fungi provides insights into their industrial applications. Notably, extensive research within the scientific community has focused on Aspergillus and Trichoderma species for the industrial production of proteins and enzymes. This review also presents practical examples of genetic engineering strategies aimed at augmenting enzyme secretion in filamentous fungi for various industrial applications. These findings underscore the potential of filamentous fungi as versatile platforms for protein production and highlight avenues for future research and technological advancement in this field.
ABSTRACT 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.
ABSTRACT The BRS Tropical rice cultivar was developed for tropical foodplains, but is susceptible to sheath blight, making the Waitea circinata bioagent an important tool in managing the disease. ...This study aimed to assess the W. circinata mechanisms involved in the parasitism and resistance induction to sheath blight. The in vitro antagonism was assessed by optical microscopy and scanning electron microscopy, followed by quantification of the lytic enzymes involved in parasitism, such as chitinase, glucanase and protease. An effect of the W. circinata mycoparasite against Rhizoctonia solani was observed. The W. circinata application suppressed the sheath blight by up to 65 % and increased the chitinase, glucanase and lipoxygenase activity 72 h after the inoculation and that of the peroxidase 96 h after the inoculation. Thus, W. circinata showed to be efficient in suppressing sheath blight by parasitism and induced resistance via the activation of biochemical mechanisms.
RESUMO A cultivar de arroz BRS Tropical foi desenvolvida para várzeas tropicais, mas é suscetível à queima da bainha, tornando o bioagente Waitea circinata uma importante ferramenta no manejo da doença. Objetivou-se avaliar os mecanismos de W. circinata envolvidos no parasitismo e na indução de resistência à queima da bainha. O antagonismo in vitro foi avaliado por microscopia óptica e microscopia eletrônica de varredura, seguida da quantificação das enzimas líticas envolvidas no parasitismo, como quitinase, glucanase e protease. Observou-se efeito do micoparasita W. circinata contra Rhizoctonia solani. A aplicação de W. circinata suprimiu a queima da bainha em até 65 % e aumentou a atividade da quitinase, glucanase e lipoxigenase 72 h após a inoculação e a da peroxidase 96 h após a inoculação. Assim, W. circinata mostrou-se efficiente em suprimir a queima da bainha por parasitismo e induziu resistência por meio da ativação de mecanismos bioquímicos.
Magnaporthe oryzae (Mo) damages rice leaf, and Thichoderma asperellum (Ta) is a potential biological agent against rice blast. The aims were to identify a reduction of leaf blast severity (LBS), ...induction of biochemical defense, growth promotion, physiological changes, and modulation of four genes, promoted by four Ta isolates and Ta pool (mixed four Ta isolates). Assay 1 (A1) tested Ta, applied by soil drenched (D) or foliar spraying (S), and evaluated LBS and enzyme activity after challenge inoculation (ChI). Assay 2 (A2) detected growth promotion and physiological changes (A, E, gs, and WUE) when Ta was applied by seed treatment (ST). In assay 3 (A3), plants were treated with Ta by D and S and evaluated after ChI for physiological changes, total chlorophyll content, antioxidant enzymes, and gene expression. In A1, all treatments suppressed leaf blast, and Ta (Ufra.T06) promoted CHI, GLU, LOX, and PAL activities in all applications form. In A2, the Ta pool promoted increases of 52%, 12,24%, 90%, and 40% in A, E, gs, and WUE, respectively. In A3, the Ta pool suppressed LBS by 89% and modulated J10sPR10 and LOX-RLL expression by increasing 35% and 474%, respectively. Ta pool is a stable combination for a multiple function biological agent and should be inserted in integrated rice management.
•T. asperellum suppressed rice blast with different applications forms•T. asperellum modulates defense genes during responses to M. oryzae•Positive growth promotion and physiological changes was detected in rice plants treated by T. asperellum•T. asperellum promoted CHI, GLU, LOX, and PAL activities in all applications form
Rhizobacteria are valuable sources of compounds that can be used for the integrated management of diseases in rice. Here, we aimed to explore the metabolism and organize and annotate the metabolites ...of
Bacillus
sp. and
Serratia marcescens
isolates using molecular networking and evaluate their fungicidal effects against
Magnaporthe oryzae
and
Bipolaris oryzae.
We obtained bacterial extracts after 6 and 16-h incubation via liquid–liquid extraction using ethyl acetate as solvent. We performed UHPLC-MS analysis and data processing using molecular networking and conducted biological assays in rice plants. Using the Global Natural Product Social spectral libraries, we annotated the following compounds: austinoneol, Phe-Pro,
N
-acetyl-
l
-leucine, Leu-Gly, Ile-Leu, Phe-Pro, 2,5-piperazinedione, 3-(1H-indol-3-methyl)-6-methyl-cyclo(
d
-Trp-
l
-Pro), and cholic acid. Results of the biological assays showed that the bacterial extracts reduced the mycelial growth of both pathogens in all treatments compared to the control. In the greenhouse setup, 8 days after the challenge for leaf gray spot and leaf blast, all treatments affected up to 4.4% of the leaf area, with an area under disease progress curve of 13.24, showing significant difference compared to the control, which affected 23% of the leaf area, with an AUDPC of 44.65. Our study provides potential new sources of natural products to be applied in the integrated management of rice.
The goal of the present study was to characterize anatomical and biochemical changes in rice plant roots in response to seed treatment with rhizobacteria (Burkholderia pyrrocinia (R-46) + Pseudomonas ...fluorescens (R-55)) and Trichoderma asperellum (Ta: mixture of strains T-06, T-09, T-12, and T-52). The experimental design was completely randomized, with six treatments (R-46, R-55, R-46 + R-55, Ta+ R-46 + R-55, Ta, and control) and ten replicates. Treatments Ta and R-46 + R-55 increased the root length and diameter as well as the cortex expansion and induced a 2% expansion of the aerenchymal space. Treatments Ta and R-46 increased the vascular cylinder diameter. The number of protoxylem poles and metaxylem vessel elements was increased by R-46 and R-55. The total phenol content increased with treatments Ta, R-46 + R-55, R-46, and R-55, and all the treatments increased the flavonoid content. The lignin content increased with the Ta and R-55 treatments. All the root architecture modifications resulting from the interaction between seedlings and bioagents (rhizobacteria and Trichoderma spp.) observed in the present study favored the root plasticity of rice seedlings.
A inoculação e coinoculação do arroz de terras altas com rizobactérias multifuncionais pode promover o crescimento das plantas, especialmente do sistema radicular. Assim, este estudo teve como ...objetivo avaliar o efeito da inoculação e coinoculação com Azospirillum sp. e Bacillus sp. no desenvolvimento inicial do arroz de terras altas. O experimento foi conduzido em delineamento inteiramente casualizado com 4 tratamentos e 10 repetições, totalizando 40 parcelas. Os tratamentos foram: 1) Ab-V5 (Azospirillum brasilense), 2) BRM 63573 (Bacillus sp.), 3) coinoculação de Ab-V5 + BRM 63573 e 4) controle (sem rizobactérias). A inoculação e coinoculação com as rizobactérias multifuncionais Ab-V5 e BRM 63573 proporcionaram efeitos positivos no desenvolvimento inicial do arroz de terras altas. A inoculação com o isolado BRM 63573 teve efeitos significativos no comprimento da raiz, parte aérea e biomassa total, enquanto a inoculação com o isolado Ab-V5 teve efeitos significativos no comprimento da raiz e produção de raiz e biomassa total. O tratamento com coinoculação teve efeitos significativos em variáveis como diâmetro, volume e superfície total de raiz e biomassa de raiz e total. O tratamento controle (sem rizobactérias multifuncionais) apresentou os piores resultados para a maioria das variáveis analisadas.
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