Caracterização do extrato de arruda e avaliação de seu potencial para o controle da brusone

O objetivo deste trabalho foi purificar e padronizar o extrato de arruda (Ruta graveolens) e avaliar o seu efeito sobre Magnaporthe oryzae como alternativa para a gestão integrada da brusone. A droga vegetal foi caracterizada, o extrato líquido obtido, e a metodologia para quantificar os marcadores padrão psoraleno e bergapteno validada. O extrato de arruda e os marcadores, isolados ou combinados, foram ensaiados in vitro, assim como em condições de casa de vegetação, para a avaliação de sua capacidade de suprimir a brusone, pela avaliação do crescimento micelial, da germinação de conídios e da formação de apressório. O extrato de arruda inibiu o crescimento micelial de M. oryzae (100%), a germinação de conídios (DL₅₀=0,237 mg) e a formação de apressórios (DL₅₀=0,121 mg); além disso, o extrato reduziu a severidade da brusone em 80,84%. A microscopia de fluorescência mostrou que o extrato de arruda não danificou a parede celular e a membrana plasmática de M. oryzae, o que indica outro modo de ação. O extrato de arruda apresenta grande potencial para o controle da brusone em folhas de arroz.

Termos para indexação:
Magnaporthe oryzae; Ruta graveolens; bergapteno; psoraleno

Autoria

Karinna Bannach Reis

Universidade Federal de Goiás, Faculdade de Farmácia, Laboratório de Pesquisas de Produtos Naturais, Caixa Postal 131, CEP 74001-970 Goiânia, GO, Brazil. E-mail: kaka.bch@gmail.com, pjrpaula@gmail.com, farmacotecnicaufg@yahoo.com.brUniversidade Federal de GoiásBrazilGoiânia, GO, BrazilUniversidade Federal de Goiás, Faculdade de Farmácia, Laboratório de Pesquisas de Produtos Naturais, Caixa Postal 131, CEP 74001-970 Goiânia, GO, Brazil. E-mail: kaka.bch@gmail.com, pjrpaula@gmail.com, farmacotecnicaufg@yahoo.com.br

Marcio Vinicius de Carvalho Barros Côrtes

Embrapa Arroz e Feijão, Rodovia GO-462, Km 12, Zona Rural, Caixa Postal 179, CEP 75375-000 Santo Antônio de Goiás, GO, Brazil. E-mail: marcio.cortes@embrapa.br, cristina.filippi@embrapa.brEmbrapa Arroz e FeijãoBrazilSanto Antônio de Goiás, GO, BrazilEmbrapa Arroz e Feijão, Rodovia GO-462, Km 12, Zona Rural, Caixa Postal 179, CEP 75375-000 Santo Antônio de Goiás, GO, Brazil. E-mail: marcio.cortes@embrapa.br, cristina.filippi@embrapa.br

Frederico Severino Martins

Universidade de São Paulo, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, CEP 14040-903 Ribeirão Preto, SP, Brazil. E-mail: fsmfarmacia@hotmail.comUniversidade de São PauloBrazilRibeirão Preto, SP, BrazilUniversidade de São Paulo, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, CEP 14040-903 Ribeirão Preto, SP, Brazil. E-mail: fsmfarmacia@hotmail.com

Marta Cristina Corsi de Filippi

José Realino de Paula

Edemilson Cardoso da Conceição

SCIMAGO INSTITUTIONS RANKINGS

Figuras | Tabelas

Figuras (7)
Tabelas (1)

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Figure 1:
Inhibition of Magnaporthe oryzae (mm) mycelial growth, after standardized Ruta graveolens extract treatment at different concentrations (0, 3.38, 6.76, 13.52, 27.04, 40.56, 54.08, 67.61 mg mL^-1). Means followed by equal letters do not differ significantly, according to Duncan's test, at 5% probability

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Figure 2:
Petri dishes showing inhibition of Magnaporthe oryzae mycelial growth, after standardized Ruta graveolens extract treatment at different concentrations (mg mL^-1): A, 0; B, 3.38; C, 6.76; D, 13.52; E, 27.04; F, 40.56; G, 54.08; H, 67.61.

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Figure 3:
Germination of Magnaporthe oryzae conidia. A, concentration of standardized Ruta graveolens extract (mg): T1, 0.0; T2, 0.10; T3: 0.20; T4: 0.30; T5 0.40; T6: 0.51; T7: 0.61; T8: 0.71; T9: 0.81; T10: 0.91; T11: 1.01. Columns followed by equal letters did not differ, according to Duncan's test, at 5% probability. B, effect of Ruta graveolens extract concentrations on the inhibition of conidial germination. Probit - log dosage curve.

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Figure 4:
Magnaporthe oryzae appressorium formation. A, concentration of standardized Ruta graveolens extract (mg): T1, 0.0; T2, 0.10; T3, 0.20; T4, 0.30; T5, 0.40; T6, 0.51; T7, 0.61; T8, 0.71; T 9, 0.81; T10, 0.91; T11, 1.01. Columns followed by equal letters did not differ significantly, according to Duncan's test, at 5% probability. B, effects of Ruta graveolens extract concentrations on appressorium inhibition. Probit - log dosage curve.

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Figure 5:
Germination and formation of appressorium. Magnaporthe oryzae conidia mixed with 7.09 mg (A), 0.71 mg (B) and 0.10 mg (C) of Ruta graveolens extract after 48 hours of deposition on a hydrophobic surface under optical microscope. Magnaporthe oryzae conidia mixed with water (D, control) and 0.304 mg (E) of Ruta graveolens extract after 72 hours of deposition on a hydrophobic surface, added of calcofluor white reagent. The image was obtained using a fluorescent microscope at 400X magnification.

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Figure 6:
Disease severity (A) and the blast lesion type (B) on rice leaves. Treatments: 1, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia; 2, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia + plant extract; 3, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia +18.26 μg mL^-1 of psoralen; 4, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia + 29.14 μg mL^-1 of bergapten; 5, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia + 18.26 μg mL^-1 of psoralen + 29.14 μg mL^-1 of bergapten. Means followed by equal letters did not differ, according to Duncan's test, at 5% probability

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Figure 7:
Sample of rice leaves representing how disease severity (A) and blast lesion type (B) developed. Treatments: 1, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia; 2, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia + plant extract; 3, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia + 18.26 μg mL^-1 of psoralen; 4, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia + 29.14 μg mL^-1 of bergapten; 5, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia + 18.26 μg mL^-1 of psoralen + 29.14 μg mL^-1 of bergapten.

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Table 1:
Treatments of blast disease assessment under greenhouse conditions.

Figure 1: Inhibition of Magnaporthe oryzae (mm) mycelial growth, after standardized Ruta graveolens extract treatment at different concentrations (0, 3.38, 6.76, 13.52, 27.04, 40.56, 54.08, 67.61 mg mL^-1). Means followed by equal letters do not differ significantly, according to Duncan's test, at 5% probability

Figure 2: Petri dishes showing inhibition of Magnaporthe oryzae mycelial growth, after standardized Ruta graveolens extract treatment at different concentrations (mg mL^-1): A, 0; B, 3.38; C, 6.76; D, 13.52; E, 27.04; F, 40.56; G, 54.08; H, 67.61.

Figure 3: Germination of Magnaporthe oryzae conidia. A, concentration of standardized Ruta graveolens extract (mg): T1, 0.0; T2, 0.10; T3: 0.20; T4: 0.30; T5 0.40; T6: 0.51; T7: 0.61; T8: 0.71; T9: 0.81; T10: 0.91; T11: 1.01. Columns followed by equal letters did not differ, according to Duncan's test, at 5% probability. B, effect of Ruta graveolens extract concentrations on the inhibition of conidial germination. Probit - log dosage curve.

Figure 4: Magnaporthe oryzae appressorium formation. A, concentration of standardized Ruta graveolens extract (mg): T1, 0.0; T2, 0.10; T3, 0.20; T4, 0.30; T5, 0.40; T6, 0.51; T7, 0.61; T8, 0.71; T 9, 0.81; T10, 0.91; T11, 1.01. Columns followed by equal letters did not differ significantly, according to Duncan's test, at 5% probability. B, effects of Ruta graveolens extract concentrations on appressorium inhibition. Probit - log dosage curve.

Figure 5: Germination and formation of appressorium. Magnaporthe oryzae conidia mixed with 7.09 mg (A), 0.71 mg (B) and 0.10 mg (C) of Ruta graveolens extract after 48 hours of deposition on a hydrophobic surface under optical microscope. Magnaporthe oryzae conidia mixed with water (D, control) and 0.304 mg (E) of Ruta graveolens extract after 72 hours of deposition on a hydrophobic surface, added of calcofluor white reagent. The image was obtained using a fluorescent microscope at 400X magnification.

Figure 6: Disease severity (A) and the blast lesion type (B) on rice leaves. Treatments: 1, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia; 2, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia + plant extract; 3, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia +18.26 μg mL^-1 of psoralen; 4, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia + 29.14 μg mL^-1 of bergapten; 5, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia + 18.26 μg mL^-1 of psoralen + 29.14 μg mL^-1 of bergapten. Means followed by equal letters did not differ, according to Duncan's test, at 5% probability

Figure 7: Sample of rice leaves representing how disease severity (A) and blast lesion type (B) developed. Treatments: 1, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia; 2, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia + plant extract; 3, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia + 18.26 μg mL^-1 of psoralen; 4, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia + 29.14 μg mL^-1 of bergapten; 5, 3x10⁵ mL^-1 of Magnaporthe oryzae conidia + 18.26 μg mL^-1 of psoralen + 29.14 μg mL^-1 of bergapten.

Table 1: Treatments of blast disease assessment under greenhouse conditions.

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Caracterização do extrato de arruda e avaliação de seu potencial para o controle da brusone

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