Mechanisms involved in the biocontrol of rice sheath blight by <i>Waitea circinata</i>

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.

KEYWORDS:
Oryza sativa ; Rhizoctonia solani ; controle biológico

Authorship

Kellen Cristhina Inácio Sousa

Universidade Federal de Goiás, Instituto de Ciências Biológicas, Departamento de Genética, Goiânia, GO, Brasil. E-mail/ORCID: bio.kcisbr@gmail.com/0000-0002-5512-855X; amandachaibub@gmail.com/0000-0002-4025-668X; jacquelinecamposcarvalho@gmail.com/0000-0001-5393-9087; leila_garces_araujo@ufg.br/0000-0002-3238-4999Universidade Federal de GoiásBrasilGoiânia, GO, BrasilUniversidade Federal de Goiás, Instituto de Ciências Biológicas, Departamento de Genética, Goiânia, GO, Brasil. E-mail/ORCID: bio.kcisbr@gmail.com/0000-0002-5512-855X; amandachaibub@gmail.com/0000-0002-4025-668X; jacquelinecamposcarvalho@gmail.com/0000-0001-5393-9087; leila_garces_araujo@ufg.br/0000-0002-3238-4999

http://orcid.org/0000-0002-5512-855X

Amanda Abdallah Chaibub

http://orcid.org/0000-0002-4025-668X

Jacqueline Campos Borba de Carvalho

http://orcid.org/0000-0001-5393-9087

Marta Cristina Corsi de Filippi

Empresa Brasileira de Pesquisa Agropecuária (Embrapa Arroz e Feijão), Santo Antônio de Goiás, GO, Brasil. E-mail/ORCID: cristina.flippi@embrapa.br/0000-0003-1676-8164Empresa Brasileira de Pesquisa Agropecuária (Embrapa Arroz e Feijão)BrasilSanto Antônio de Goiás, GO, BrasilEmpresa Brasileira de Pesquisa Agropecuária (Embrapa Arroz e Feijão), Santo Antônio de Goiás, GO, Brasil. E-mail/ORCID: cristina.flippi@embrapa.br/0000-0003-1676-8164

http://orcid.org/0000-0003-1676-8164

Leila Garcês de Araújo

http://orcid.org/0000-0002-3238-4999

SCIMAGO INSTITUTIONS RANKINGS

Figures

Figures (7)

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Figure 1
In vitro interaction between Waitea circinata and Rhizoctonia solani: A) R. solani in PDA culture medium; B) W. circinata in PDA culture medium; C) R. solani pathogen (P) and W. circinata mycorrhizal fungus (M) paired in Petri dishes; D) R. solani hyphae; E) W. circinata hyphae (white arrow) coiled around R. solani hyphae (red arrow) under an optical microscope; and F) under a scanning electron microscope. Bars: 10 µm.

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Figure 2
In vitro enzymatic activity of Waitea circinata in co-culture with Rhizoctonia solani: A) glucanase; B) chitinase; C) protease. * Means significantly different, according to the Tukey test (p < 0.05). Bars indicate the mean standard error.

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Figure 3
Evaluation of Waitea circinata pathogenicity in rice plants: A) plants sprayed only with water; B) plants sprayed with 5 g L^-1; C) plants sprayed with 10 g L^-1; D) plants inoculated with 9-mm disks of W. circinata applied to the soil.

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Figure 4
Sheath blight severity (A, C and E) and area under the disease progress curve - AUDPC (B, D and F) for the greenhouse assays (A1: A-B; A2: C-D; A3: E-F) with Waitea circinata applied beforehand (disks) or simultaneously (sprayed with 5 and 10 g L^-1).

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Figure 5
In vivo interaction between Waitea circinata and Rhizoctonia solani, with rice plants in a greenhouse (left) and respective scanning electron microscopy (right): A) plants sprayed only with autoclaved distilled water; B) plants treated with W. circinata soil disks (20 days after sowing - DAS); C-D) plants inoculated only with R. solani at 55 DAS, large number of hyphae on leaves (C), sheath degraded by the pathogen (D); E-F) plants treated with W. circinata disks at 20 DAS and subsequently inoculated with R. solani at 55 DAS. Arrows: A-B) glandular trichomes in the guard cells closed the stomata; D) necrotic sheaths with mycelial growth of the pathogen; E) waxy layer; F) crystals. Bars: 10 µm (A, B, E, F) and 50 µm (C, D).

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Figure 6
In vivo enzymatic activity [A) chitinase; B) glucanase; C) lipoxygenase; D) phenylalanine ammonia-lyase; E) peroxidase] in rice plants treated with Waitea circinata and subsequently inoculated with the pathogen Rhizoctonia solani (R. solani x W. circinata), treated with W. circinata alone and inoculated only with the pathogen (R. solani).

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Figure 7
Multivariate analysis of enzymatic activity and disease severity for plants treated only with Waitea circinata, inoculated with Rhizoctonia solani alone or treated with W. circinata and inoculated with R. solani: A) Pearson’s linear correlation, with Bonferroni correction, between chitinase (CHI), glucanase (GLU), lipoxygenase (LOX), phenylalanine ammonia-lyase (PAL) and peroxidase (POX) activity and severity; B) principal component analysis of the studied variables (enzymes: CHI, GLU, LOX, PAL and POX and severity).

Figure 1 In vitro interaction between Waitea circinata and Rhizoctonia solani: A) R. solani in PDA culture medium; B) W. circinata in PDA culture medium; C) R. solani pathogen (P) and W. circinata mycorrhizal fungus (M) paired in Petri dishes; D) R. solani hyphae; E) W. circinata hyphae (white arrow) coiled around R. solani hyphae (red arrow) under an optical microscope; and F) under a scanning electron microscope. Bars: 10 µm.

Figure 2 In vitro enzymatic activity of Waitea circinata in co-culture with Rhizoctonia solani: A) glucanase; B) chitinase; C) protease. * Means significantly different, according to the Tukey test (p < 0.05). Bars indicate the mean standard error.

Figure 3 Evaluation of Waitea circinata pathogenicity in rice plants: A) plants sprayed only with water; B) plants sprayed with 5 g L^-1; C) plants sprayed with 10 g L^-1; D) plants inoculated with 9-mm disks of W. circinata applied to the soil.

Figure 4 Sheath blight severity (A, C and E) and area under the disease progress curve - AUDPC (B, D and F) for the greenhouse assays (A1: A-B; A2: C-D; A3: E-F) with Waitea circinata applied beforehand (disks) or simultaneously (sprayed with 5 and 10 g L^-1).

Figure 5 In vivo interaction between Waitea circinata and Rhizoctonia solani, with rice plants in a greenhouse (left) and respective scanning electron microscopy (right): A) plants sprayed only with autoclaved distilled water; B) plants treated with W. circinata soil disks (20 days after sowing - DAS); C-D) plants inoculated only with R. solani at 55 DAS, large number of hyphae on leaves (C), sheath degraded by the pathogen (D); E-F) plants treated with W. circinata disks at 20 DAS and subsequently inoculated with R. solani at 55 DAS. Arrows: A-B) glandular trichomes in the guard cells closed the stomata; D) necrotic sheaths with mycelial growth of the pathogen; E) waxy layer; F) crystals. Bars: 10 µm (A, B, E, F) and 50 µm (C, D).

Figure 6 In vivo enzymatic activity [A) chitinase; B) glucanase; C) lipoxygenase; D) phenylalanine ammonia-lyase; E) peroxidase] in rice plants treated with Waitea circinata and subsequently inoculated with the pathogen Rhizoctonia solani (R. solani x W. circinata), treated with W. circinata alone and inoculated only with the pathogen (R. solani).

Figure 7 Multivariate analysis of enzymatic activity and disease severity for plants treated only with Waitea circinata, inoculated with Rhizoctonia solani alone or treated with W. circinata and inoculated with R. solani: A) Pearson’s linear correlation, with Bonferroni correction, between chitinase (CHI), glucanase (GLU), lipoxygenase (LOX), phenylalanine ammonia-lyase (PAL) and peroxidase (POX) activity and severity; B) principal component analysis of the studied variables (enzymes: CHI, GLU, LOX, PAL and POX and severity).

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Mechanisms involved in the biocontrol of rice sheath blight by <i>Waitea circinata</i>

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