Nep1‐like protein from <i>Moniliophthora perniciosa</i> induces a rapid proteome and metabolome reprogramming in cells of <i>Nicotiana benthamiana</i>

Dias, Cristiano Villela; Camillo, Luciana; Oliveira, Guilherme A. P. de; Sena, Jamilly Azevedo Leal; Santiago, André S.; Sousa, Sanderson Tarciso Pereira de; Mendes, Juliano S.; Pirovani, Carlos Priminho; Alvim, Fátima Cerqueira; Costa, Márcio Gilberto Cardoso

doi:10.1111/ppl.12061

Cited by 20 publications

(18 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Protein identification was performed using a Mascot server (Version 2.1.04, Matrix Science) and the T. cacao genome database [68] . The search criteria used were trypsin digestion, variable modifications set as cysteine (Cys) and methionine (Met), max of one missed cleavages allowed and peptide mass tolerance of ±0.3 Da for the parent ion and 0.10 Da for the fragment ions [66] , [67] .…”

Section: Methodsmentioning

confidence: 99%

Analysis of Gene Expression and Proteomic Profiles of Clonal Genotypes from Theobroma cacao Subjected to Soil Flooding

2014

Self Cite

View full text Add to dashboard Cite

Soil flooding causes changes in gene transcription, synthesis and degradation of proteins and cell metabolism. The main objective of this study was to understand the biological events of Theobroma cacao during soil flooding-induced stress, using the analyses of gene expression and activity of key enzymes involved in fermentation, as well as the identification of differentially expressed proteins by mass spectrometry in two contrasting genotypes for flooding tolerance (tolerant - TSA-792 and susceptible - TSH-774). Soil anoxia caused by flooding has led to changes in the expression pattern of genes associated with the biosynthesis of alcohol dehydrogenase (ADH), pyruvate decarboxylase (PDC) and lactate dehydrogenase (LDH) in leaves and roots of the two evaluated genotypes. Significant differences were observed between the enzyme activities of the two genotypes. Leaves and roots of the TSA-792 genotype showed higher ADH activity as compared to the TSH-774 genotype, whereas the activities of PDC and LDH have varied over the 96 h of soil flooding, being higher for TSA-792 genotype, at the initial stage, and TSH-774 genotype, at the final stage. Some of the identified proteins are those typical of the anaerobic metabolism-involved in glycolysis and alcoholic fermentation-and different proteins associated with photosynthesis, protein metabolism and oxidative stress. The ability to maintain glycolysis and induce fermentation was observed to play an important role in anoxia tolerance in cacao and may also serve to distinguish tolerant and susceptible genotypes in relation to this stressor.

show abstract

Section: Methodsmentioning

confidence: 99%

Analysis of Gene Expression and Proteomic Profiles of Clonal Genotypes from Theobroma cacao Subjected to Soil Flooding

2014

Self Cite

View full text Add to dashboard Cite

show abstract

“…This indicates interference in the pathosystem because of overexpression of cystatins, and may interfere with symptom development and disease progression, which is directly related to cellular death induced by MpNEP2. Villela-Dias et al (2014) observed responses similar to necrosis in cell cultures of N. benthamiana (expressing green fluorescent protein) treated with MpNEP2, which is characterized by swelling of organelles and plasma membrane rupture among other processes, indicating direct damage to the plasma membrane. Studies by Mandal et al (2006) confirmed that cystatin expression can neutralize the effect of fungal proteases and block PCD during pathogenesis of the fungus.…”

Section: Discussionmentioning

confidence: 85%

“…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%

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

Santana¹,

Costa²,

Pirovani³

et al. 2014

Genet. Mol. Res.

View full text Add to dashboard Cite

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.

show abstract

“…were excised from gels and placed in microtubes. Then, they were bleached and subjected to trypsin digestion [ 23 ]. Next, the samples containing recovered peptides were vacuum concentrated until they reached a volume of 10–15 μL.…”

Section: Methodsmentioning

confidence: 99%

“…The resulting peptides from tryptic digestion were subjected to liquid chromatography with tandem mass spectrometry (LC-MS/MS) on a nanoAcquity system (Waters, Milford, MA) coupled with a Q-ToF micro mass spectrometer (Waters), according to methods described elsewhere [ 23 ]. The raw data were processed, and the resulting spectra were analyzed in the ProteinLynx Global Server 4.2 software (Waters) and compared with the SwissProt database ( http://www.uniprot.org/downloads , October 2011).…”

Section: Methodsmentioning

confidence: 99%

Proteomic analysis during of spore germination of Moniliophthora perniciosa, the causal agent of witches’ broom disease in cacao

et al. 2017

View full text Add to dashboard Cite

Background Moniliophthora perniciosa is a phytopathogenic fungus responsible for witches’ broom disease of cacao trees (Theobroma cacao L.). Understanding the molecular events during germination of the pathogen may enable the development of strategies for disease control in these economically important plants. In this study, we determined a comparative proteomic profile of M. perniciosa basidiospores during germination by two-dimensional SDS-PAGE and mass spectrometry.ResultsA total of 316 proteins were identified. Molecular changes during the development of the germinative tube were identified by a hierarchical clustering analysis based on the differential accumulation of proteins. Proteins associated with fungal filamentation, such as septin and kinesin, were detected only 4 h after germination (hag). A transcription factor related to biosynthesis of the secondary metabolite fumagillin, which can form hybrids with polyketides, was induced 2 hag, and polyketide synthase was observed 4 hag. The accumulation of ATP synthase, binding immunoglobulin protein (BiP), and catalase was validated by western blotting.ConclusionsIn this study, we showed variations in protein expression during the early germination stages of fungus M. perniciosa. Proteins associated with fungal filamentation, and consequently with virulence, were detected in basidiospores 4 hag., for example, septin and kinesin. We discuss these results and propose a model of the germination of fungus M. perniciosa. This research can help elucidate the mechanisms underlying basic processes of host invasion and to develop strategies for control of the disease.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-017-1085-4) contains supplementary material, which is available to authorized users.

show abstract

Nep1‐like protein from Moniliophthora perniciosa induces a rapid proteome and metabolome reprogramming in cells of Nicotiana benthamiana

Cited by 20 publications

References 54 publications

Analysis of Gene Expression and Proteomic Profiles of Clonal Genotypes from Theobroma cacao Subjected to Soil Flooding

Analysis of Gene Expression and Proteomic Profiles of Clonal Genotypes from Theobroma cacao Subjected to Soil Flooding

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

Proteomic analysis during of spore germination of Moniliophthora perniciosa, the causal agent of witches’ broom disease in cacao

Contact Info

Product

Resources

About