Characterization of necrosis and ethylene-inducing proteins (NEP) in the basidiomycete Moniliophthora perniciosa, the causal agent of witches' broom in Theobroma cacao

García, Odalys Águila; Macêdo, Joci Neuby Alves; Tiburcio, Ricardo Augusto; Zaparoli, G.; Rincones, Johana; Bittencourt, Livia Maria Cabral; Ceita, Geruza Oliveira; Micheli, Fabienne; Gesteira, Abelmon S.; Mariano, Andrea; Schiavinato, Marlene Aparecida; Medrano, Francisco J.; Meinhardt, Lyndel W.; Pereira, Gonçalo Amarante Guimarães; Cascardo, Júlio Cézar M.

doi:10.1016/j.mycres.2007.01.017

Cited by 80 publications

(88 citation statements)

References 49 publications

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“…Total proteins were isolated from a mix of 20 T. cacao meristems, 3 days after infiltration with either a GLY-grown M. perniciosa secretome, Mp-NeP2 (M. perniciosa necrosis-inducing protein; Garcia et al, 2007), PB (10 mM, pH 5.5) or a suspension of M. perniciosa basidiospores. The meristems were frozen in liquid nitrogen and immediately submitted to protein extraction according to Pirovani et al (2008).…”

Section: T Cacao Meristem Infiltration and Induction Of Binding Proteinmentioning

confidence: 99%

Carbon source-induced changes in the physiology of the cacao pathogen Moniliophthora perniciosa (Basidiomycetes) affect mycelial morphology and secretion of necrosis-inducing proteins

Alvim¹,

Mattos²,

Pirovani³

et al. 2009

Genet. Mol. Res.

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ABSTRACT. Quantitative and qualitative relationships were found between secreted proteins and their activity, and the hyphal morphology of Moniliophthora perniciosa, the causal agent of witches' broom disease in Theobroma cacao. This fungus was grown on fermentable and non-fermentable carbon sources; significant differences in mycelial morphology were observed and correlated with the carbon source. A biological assay performed with Nicotiana tabacum leaves revealed that the necrosis-related activity of extracellular fungal proteins also differed with carbon source. There were clear differences in the type and quantity of the secreted proteins. In addition, the expression of the cacao molecular chaperone BiP increased after treatment with secreted proteins, suggesting a physiological response to the fungus secretome. We suggest that the carbon source-dependent energy metabolism of M. perniciosa results in physiological alterations in protein expression and secretion; these may affect not only M. perniciosa growth, but also its ability to express pathogenicity proteins.

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Section: T Cacao Meristem Infiltration and Induction Of Binding Proteinmentioning

confidence: 99%

Carbon source-induced changes in the physiology of the cacao pathogen Moniliophthora perniciosa (Basidiomycetes) affect mycelial morphology and secretion of necrosis-inducing proteins

Alvim¹,

Mattos²,

Pirovani³

et al. 2009

Genet. Mol. Res.

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“…2 Since then, the production of this commodity has severely dropped resulting in the shift of Brazil from the second largest cacao exporter to a cacao importer. 3 To make matters worse, this disease has caused social and environmental damage to the region as the local producers are devastating the local tropical forest to sow alternative crops. In order to minimize WBD impact, several control programs have been developed.…”

Section: Introductionmentioning

confidence: 99%

In vitro screening and chemometrics analysis on a series of azole derivatives with fungicide activity against moniliophthora perniciosa

Mota¹,

Barros²,

Castilho³

2010

J. Braz. Chem. Soc.

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Moniliophthora perniciosa, o agente causal da vassoura-de-bruxa do cacaueiro, diminuiu significativamente a produção de cacau, especialmente no estado da Bahia, a maior região produtora de cacau do continente americano. As formas de controle desenvolvidas até o momento têm baixa eficiência. Derivados de azol são ativos tanto in vitro quanto in loco contra M. perniciosa, porém não há um estudo sistemático sobre a atividade dos azóis contra este fitopatogeno. Dados biológicos, obtidos em um ensaio padronizado, foram utilizados para criação de modelos quimiométricos, que destacam características físico e estruturais importantes para a atividade fungicida de derivados de azol frente a M. perniciosa. De acordo com os modelos de PCA e SIMCA, características eletrônicas, representadas pelos descritores BEHe3 e JGI4, paralelamente à possibilidadde de realizar ligações de H e ausência de átomos de cloro, distantes de 6-8 ligações dos nitrogênios do anel azólico, parecem contribuir para atividade fungicida dos compostos estudados Moniliophthora perniciosa, the causal agent of witches' broom disease in Theobroma cacao, significantly decreased cacao production, especially in Bahia State, the largest cocoa producing of the American continent. Control programs developed so far have low efficiency. Azole derivatives are active both in vitro and in loco against M. perniciosa, however there is no comprehensive study on the activity of azoles against this phytopatogen. Standardized in vitro biological data were employed to develop supervised and unsupervised chemometric models that highlight physicochemical and structural features that are crucial for azole's fungicidal activity against M. perniciosa. Thus, PCA and SIMCA models suggest that electronegativity (BEHe3) and dipolar moment (JGI4), as well as H-bonding to M. pernciosa's lanosterol 14α-desmethylase active site and lack of Cl atoms 6 to 8 bonds from the azole's nitrogen atoms play a major role to azoles' fungicide activity.

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“…The recombinant protein MpNEP2 that had been expressed and purified from the clone produced by Garcia et al (2007) at a 0.2 mM concentration was injected into the foliar mesophyll of Tccys4:tobacco lines and controls (wt) using an insulin syringe. The leaves used were fully expanded and mature (20 days after transfer).…”

Section: Tobacco Plant Response Expressing Cystatin To Mpnep2mentioning

confidence: 99%

“…To determine whether overexpression of cocoa cystatin in tobacco plants affects MpNEP2 protein activity, recombinant protein was produced from a clone prepared by Garcia et al (2007). The MpNEP2 recombinant protein, which has been purified previously, was injected into the foliar mesophyll of 5 wt tobacco plants and 5 Tccys4:tobacco transgenic plants according to the method described by Garcia et al (2007).…”

Section: Effect Of Cocoa Cystatin In Response To Mpnep2 Protein In Tomentioning

confidence: 99%

“…Previous studies have demonstrated that recombinant MpNEP2 induces cell death in the leaves and tobacco suspension cell culture (Garcia et al, 2007) as well as reprograms the proteome and metabolome of Nicotiana benthamiana cells (Villela-Dias et al, 2014). In this study, we produced tobacco plants that overexpressed a cystatin gene sequence from cocoa (Tccys4).…”

Section: Introductionmentioning

confidence: 99%

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TcCYS4, a cystatin from cocoa, reduces necrosis triggered by MpNEP2 in tobacco plants

Santana¹,

Costa²,

Pirovani³

et al. 2014

Genet. Mol. Res.

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ABSTRACT. In Brazil, most cocoa bean production occurs in Southern Bahia. Witches' broom disease arrived in this area in 1989 and has since caused heavy losses in production. The disease is caused by the basidiomycete fungus Moniliophthora perniciosa, a hemibiotrophic fungus that produces the necrosis and ethylene-inducting protein (MpNEP2) during infection; this protein can activate cysteine proteases and induce programmed cell death. Cysteine proteases can be modulated by cystatin. In this study, we overexpressed TcCYS4, a cocoa cystatin, in tobacco plants and evaluated the effect on MpNEP2 in model plants. Tccys4 cDNA was cloned into the pCAMBIA 1390 vector and inserted into the tobacco plants via Agrobacterium tumefaciens. Transgene expression was analyzed by reverse transcription-quantitative PCR and Western blot analysis. Transcript and protein levels in Tcccys4:tobacco lines were 8.9-and 1.5-fold higher than in wild-type plants (wt). Tcccys4:tobacco lines showed no change in growth compared to wt plants. CO 2 net assimilation (A) increased in Tcccys4:tobacco lines compared to wt plants. Only one line showed statistically significant stomatal conductance (gs) and transpiration rate (E) changes. MpNEP2 was infiltered into the foliar mesophyll of Tcccys4:tobacco lines and wt plants, and necrotic lesions were attenuated in lines highly expressing Tccys4. Our results suggest that cocoa cystatin TcCYS4 affects MpNEP2 activity related to the progression of programmed cell death in tobacco plants. This may occur through the action of cystatin to inhibit cysteine proteases activated by MpNEP2 in plant tissues. Further studies are necessary to examine cystatin in the Theobroma cacao-M. perniciosa pathosystem.

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Characterization of necrosis and ethylene-inducing proteins (NEP) in the basidiomycete Moniliophthora perniciosa, the causal agent of witches' broom in Theobroma cacao

Cited by 80 publications

References 49 publications

Carbon source-induced changes in the physiology of the cacao pathogen Moniliophthora perniciosa (Basidiomycetes) affect mycelial morphology and secretion of necrosis-inducing proteins

Carbon source-induced changes in the physiology of the cacao pathogen Moniliophthora perniciosa (Basidiomycetes) affect mycelial morphology and secretion of necrosis-inducing proteins

In vitro screening and chemometrics analysis on a series of azole derivatives with fungicide activity against moniliophthora perniciosa

TcCYS4, a cystatin from cocoa, reduces necrosis triggered by MpNEP2 in tobacco plants

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