Evidence of Pyricularia oryzae adaptability to tricyclazole.

Pyricularia oryzae is the etiological agent of rice blast, the most destructive disease in rice crops and chemical control based on fungicide is the main method used in its management. The aim of this study was characterize pathogenicity and identify P. oryzae isolates adapted to tricyclazole. P. oryzae monosporic isolates were collected in the state of Tocantins and inoculated in international differentiating series of rice cultivars for determination of pathotypes. After, the same isolates were inoculated in the rice cultivar IRGA 424 to evaluate resistance to fungicide Bim® 750 BR (Tricyclazole - 250 g/ha) that was applied 24 and 48 hours after pathogen inoculation (hai). Leaf blast severity and infection efficiency were evaluated 9 days after inoculation (dai), latency period (2 dai) and sporulation intensity (7 dai). Nine different pathotypes were identified, predominantly as IA group. The latent period of isolates occurred between from 48 to 120 h. The application of tricyclazole, 24 hai reduced disease severity with the exception of the isolate Py 7.1. The great variability of the pathogen allowed for adaptation to this molecule and can increase its aggressiveness and should be considered to guide the integrated management of the disease.

Molecular identification of Trichoderma sp. isolates and biochemical characterization of antagonistic interaction against rice blast.

This study aimed to identify four isolates of Trichoderma sp. (Ufra.T06, Ufra.T09, Ufra.T12, and Ufra.T52) and characterize their interaction with Magnaporthe oryzae in vitro and in vivo conditions. The four isolates of Trichoderma sp. were sequenced, investigated as an antagonist against M. oryzae in five Petri plate assays, and as an inhibitor of conidial germination appressoria formation. Finally, were quantified the lytic activity of chitinase (CHI), glucanase (GLU), and protease (PRO) during co-cultivation of Trichoderma sp. and M. oryzae. In vivo, leaf blast suppression was evaluated in two assays: simultaneous and curative application. Both in vitro and in vivo assays were scanned by electron microscopy (SEM). All isolates were identified as Trichoderma asperellum. All in vitro Petri plates assays reduced M. oryzae colony growth (paired-91.18% by Ufra.T09, volatile metabolites-all isolates equally reduced, non-volatile-68.33% by Ufra.T06, thermostability-99.77% by Ufra.T52 and co-cultivate-64.25% by Ufra.T52). The filtrates and conidia suspensions for T. asperellum isolates inhibited the conidia germination and appressoria formation significantly. In co-cultivate (mycelial or cell wall), all enzymes (GLU, CHI, and PRO) and times (24, 48, and 72 h) showed increased activity. In vivo, reduced leaf blast severity until 94.64% (Ufra.T52cs) in a simultaneous and until 85% (Ufra.T09 24 and 48 hasi) in a curative application. T. asperellum isolates showed efficient control of M. oryzae by mycoparasitism, and antibiosis mechanisms were interfered with by the M. oryzae infection process.

Induction of resistance in rice plants using bioproducts produced from Burkholderia pyrrocinia BRM 32113.

Leaf blast is the main rice disease in the world causing significant losses in productivity. Blast integrate management (BIM) requires the use of genetic resistance, cultural practices, and chemical control, although for sustainable BIM, the insertion of biological agents may be the fourth component for. The objective of this work was to test three formulations of Burkholderia pyrrocinia (BRM32113) previously selected and to verify the effectiveness in resistance induction and blast control in rice. Two experiments were carried out, in a completely randomized design with three replications, in the greenhouse (E1 and E2). E1 aimed to select the best treatment for suppressing leaf blast severity and activating plant defense mechanisms. It was composed of 8 treatments: (1) formulated 11+ B. pyrrocina × Magnaporthe oryzae; (2) formulated 17+ B. pyrrocina × M. oryzae; (3) formulated 32+ B. pyrrocina × M. oryzae; (4) formulated 11 × M. oryzae; (5) B. pyrrocinia 17 × M. oryzae; (6) formulated 32 × M. oryzae; (7) B. pyrrocina × M. oryzae; (8) M. oryzae; (9) control (water). E2 aimed to investigate the effect of the best treatments, for the promotion of plant growth and suppression of leaf blast by calculating AUDPC. It was composed of 6 treatments: (1) formulated 11+ B. pyrrocina × M. oryzae; (2) formulated 32+ B. pyrrocina × M. oryzae; (3) formulated 11 × M. oryzae; (4) formulated 32 × M. oryzae; (5) B. pyrrocina × M. oryzae; (6) water. And after, we did two assays aimed to localize this biological agent after application at seed, soil, and rice plant. In E1, formulated 11+ B. pyrrocinia and 32+ formulated and B. pyrrocina were the best, suppressing leaf blast by up to 97% and providing the significant increase of the enzymes ß-1,3-glucanase, chitinase, phenylalanine ammonia lyase, lipoxygenase, and salicylic acid at 24 h and 48 h after inoculation with M. oryzae. In E2, treatments formulated 11+ B. pyrrocinia, formulated 32+ B. pyrrocinia, and B. pyrrocina provided more significant increases in growth promotion and reduced area under disease progress curve. B. pyrrocinia was detected in the rice plant for 18 days, predominantly in the root system (internal and external). The use of B. pyrrocinia formulations based on sugarcane molasses and glycerol can be an essential strategy for sustainable management. Although all the benefits come from these sustainable formulations, the adoption by commercial biological segment depends on an established formulation process. It seems that all the results showed here by this research will be readily assimilated by startups of the organic segment.

Defence responses in rice plants in prior and simultaneous applications of Cladosporium sp. during leaf blast suppression.

An alternative method to control rice blast (Magnaporthe oryzae) is to include biological agent in the disease management strategy. The objective of this study was to assess the leaf blast-suppressing effects of rice phylloplane fungi. One Cladosporium sp. phylloplane fungus was shown to possess biocontrolling traits based on its morphological characteristics and an analysis of its 18S ribosomal DNA. Experiments aimed at determining the optimal time to apply the bioagent and the mechanisms involved in its rice blast-suppressing activities were performed under controlled greenhouse conditions. We used foliar spraying to apply the Cladosporium sp. 48 h prior to applying the pathogen, and we found that this increased the enzymatic activity. Furthermore, in vitro tests performed using isolate C24 showed that it possessed the ability to secrete endoxylanases and endoglucanases. When Cladosporium sp. was applied either prior to or simultaneous with the pathogen, we observed a significant increase in defence enzyme activity, and rice blast was suppressed by 84.0 and 78.6 %, respectively. However, some enzymes showed higher activity at 24 h while others did so at 48 h after the challenge inoculation. Cladosporium sp. is a biological agent that is capable of suppressing rice leaf blast by activating biochemical defence mechanisms in rice plants. It is highly adapted to natural field conditions and should be included in further studies aimed at developing strategies to support ecologically sustainable disease management and reduce environmental pollution by the judicious use of fungicidal sprays.