Tc-cAPX, a cytosolic ascorbate peroxidase of Theobroma cacao L. engaged in the interaction with Moniliophthora perniciosa, the causing agent of witches' broom disease
The level of hydrogen peroxide (H2O2) in plants signalizes the induction of several genes, including that of ascorbate peroxidase (APX-EC 1.11.1.11). APX isoenzymes play a central role in the elimination of intracellular H2O2 and contribute to plant responses to diverse stresses. During the infection process in Theobroma cacao by Moniliophthora perniciosa oxidative stress is generated and the APX action recruited from the plant. The present work aimed to characterize the T. cacao APX involved in the molecular interaction of T. cacao-M. perniciosa. The peroxidase activity was analyzed in protein extracts from cocoa plants infected by M. perniciosa and showed the induction of peroxidases like APX in resistant cocoa plants. The cytosolic protein of T. cacao (GenBank: ABR68691.2) was phylogenetically analyzed in relation to other peroxidases from the cocoa genome and eight genes encoding APX proteins with conserved domains were also analyzed. The cDNA from cytosolic APX was cloned in pET28a and the recombinant protein expressed and purified (rTc-cAPX). The secondary structure of the protein was analyzed by Circular Dichroism (CD) displaying high proportion of ?-helices when folded. The enzymatic assay shows stable activity using ascorbate and guaiacol as an electron donor for H2O2 reduction. The pH 7.5 is the optimum for enzyme activity. Chromatographic analysis suggests that rTc-cAPX is a homodimer in solution. Results indicate that the rTc-cAPX is correctly folded, stable and biochemically active. The purified rTc-cAPX presented biotechnological potential and is adequate for future structural and functional studies.
Main Authors: | , , , , , , |
---|---|
Format: | article biblioteca |
Language: | eng |
Subjects: | F30 - Génétique et amélioration des plantes, H20 - Maladies des plantes, Theobroma cacao, relation hôte pathogène, Moniliophthora, péroxydase, résistance génétique, protéase, activité enzymatique, résistance aux organismes nuisibles, http://aims.fao.org/aos/agrovoc/c_7713, http://aims.fao.org/aos/agrovoc/c_34017, http://aims.fao.org/aos/agrovoc/c_31727, http://aims.fao.org/aos/agrovoc/c_13251, http://aims.fao.org/aos/agrovoc/c_35130, http://aims.fao.org/aos/agrovoc/c_6242, http://aims.fao.org/aos/agrovoc/c_2604, http://aims.fao.org/aos/agrovoc/c_5731, http://aims.fao.org/aos/agrovoc/c_779, |
Online Access: | http://agritrop.cirad.fr/571100/ http://agritrop.cirad.fr/571100/1/document_571100.pdf |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
id |
dig-cirad-fr-571100 |
---|---|
record_format |
koha |
spelling |
dig-cirad-fr-5711002024-01-28T21:42:40Z http://agritrop.cirad.fr/571100/ http://agritrop.cirad.fr/571100/ Tc-cAPX, a cytosolic ascorbate peroxidase of Theobroma cacao L. engaged in the interaction with Moniliophthora perniciosa, the causing agent of witches' broom disease. Rodrigues Camillo Luciana, Ribeiro Filadelfo Ciro, Monzani Paulo Sérgio, Corrêa Ronan Xavier, Peres Gramacho Karina, Micheli Fabienne, Priminho Pirovani Carlos. 2013. Plant Physiology and Biochemistry, 73 : 254-265.https://doi.org/10.1016/j.plaphy.2013.10.009 <https://doi.org/10.1016/j.plaphy.2013.10.009> Tc-cAPX, a cytosolic ascorbate peroxidase of Theobroma cacao L. engaged in the interaction with Moniliophthora perniciosa, the causing agent of witches' broom disease Rodrigues Camillo, Luciana Ribeiro Filadelfo, Ciro Monzani, Paulo Sérgio Corrêa, Ronan Xavier Peres Gramacho, Karina Micheli, Fabienne Priminho Pirovani, Carlos eng 2013 Plant Physiology and Biochemistry F30 - Génétique et amélioration des plantes H20 - Maladies des plantes Theobroma cacao relation hôte pathogène Moniliophthora péroxydase résistance génétique protéase activité enzymatique résistance aux organismes nuisibles http://aims.fao.org/aos/agrovoc/c_7713 http://aims.fao.org/aos/agrovoc/c_34017 http://aims.fao.org/aos/agrovoc/c_31727 http://aims.fao.org/aos/agrovoc/c_13251 http://aims.fao.org/aos/agrovoc/c_35130 http://aims.fao.org/aos/agrovoc/c_6242 http://aims.fao.org/aos/agrovoc/c_2604 http://aims.fao.org/aos/agrovoc/c_5731 Bahia http://aims.fao.org/aos/agrovoc/c_779 The level of hydrogen peroxide (H2O2) in plants signalizes the induction of several genes, including that of ascorbate peroxidase (APX-EC 1.11.1.11). APX isoenzymes play a central role in the elimination of intracellular H2O2 and contribute to plant responses to diverse stresses. During the infection process in Theobroma cacao by Moniliophthora perniciosa oxidative stress is generated and the APX action recruited from the plant. The present work aimed to characterize the T. cacao APX involved in the molecular interaction of T. cacao-M. perniciosa. The peroxidase activity was analyzed in protein extracts from cocoa plants infected by M. perniciosa and showed the induction of peroxidases like APX in resistant cocoa plants. The cytosolic protein of T. cacao (GenBank: ABR68691.2) was phylogenetically analyzed in relation to other peroxidases from the cocoa genome and eight genes encoding APX proteins with conserved domains were also analyzed. The cDNA from cytosolic APX was cloned in pET28a and the recombinant protein expressed and purified (rTc-cAPX). The secondary structure of the protein was analyzed by Circular Dichroism (CD) displaying high proportion of ?-helices when folded. The enzymatic assay shows stable activity using ascorbate and guaiacol as an electron donor for H2O2 reduction. The pH 7.5 is the optimum for enzyme activity. Chromatographic analysis suggests that rTc-cAPX is a homodimer in solution. Results indicate that the rTc-cAPX is correctly folded, stable and biochemically active. The purified rTc-cAPX presented biotechnological potential and is adequate for future structural and functional studies. article info:eu-repo/semantics/article Journal Article info:eu-repo/semantics/publishedVersion http://agritrop.cirad.fr/571100/1/document_571100.pdf application/pdf Cirad license info:eu-repo/semantics/restrictedAccess https://agritrop.cirad.fr/mention_legale.html https://doi.org/10.1016/j.plaphy.2013.10.009 10.1016/j.plaphy.2013.10.009 info:eu-repo/semantics/altIdentifier/doi/10.1016/j.plaphy.2013.10.009 info:eu-repo/semantics/altIdentifier/purl/https://doi.org/10.1016/j.plaphy.2013.10.009 |
institution |
CIRAD FR |
collection |
DSpace |
country |
Francia |
countrycode |
FR |
component |
Bibliográfico |
access |
En linea |
databasecode |
dig-cirad-fr |
tag |
biblioteca |
region |
Europa del Oeste |
libraryname |
Biblioteca del CIRAD Francia |
language |
eng |
topic |
F30 - Génétique et amélioration des plantes H20 - Maladies des plantes Theobroma cacao relation hôte pathogène Moniliophthora péroxydase résistance génétique protéase activité enzymatique résistance aux organismes nuisibles http://aims.fao.org/aos/agrovoc/c_7713 http://aims.fao.org/aos/agrovoc/c_34017 http://aims.fao.org/aos/agrovoc/c_31727 http://aims.fao.org/aos/agrovoc/c_13251 http://aims.fao.org/aos/agrovoc/c_35130 http://aims.fao.org/aos/agrovoc/c_6242 http://aims.fao.org/aos/agrovoc/c_2604 http://aims.fao.org/aos/agrovoc/c_5731 http://aims.fao.org/aos/agrovoc/c_779 F30 - Génétique et amélioration des plantes H20 - Maladies des plantes Theobroma cacao relation hôte pathogène Moniliophthora péroxydase résistance génétique protéase activité enzymatique résistance aux organismes nuisibles http://aims.fao.org/aos/agrovoc/c_7713 http://aims.fao.org/aos/agrovoc/c_34017 http://aims.fao.org/aos/agrovoc/c_31727 http://aims.fao.org/aos/agrovoc/c_13251 http://aims.fao.org/aos/agrovoc/c_35130 http://aims.fao.org/aos/agrovoc/c_6242 http://aims.fao.org/aos/agrovoc/c_2604 http://aims.fao.org/aos/agrovoc/c_5731 http://aims.fao.org/aos/agrovoc/c_779 |
spellingShingle |
F30 - Génétique et amélioration des plantes H20 - Maladies des plantes Theobroma cacao relation hôte pathogène Moniliophthora péroxydase résistance génétique protéase activité enzymatique résistance aux organismes nuisibles http://aims.fao.org/aos/agrovoc/c_7713 http://aims.fao.org/aos/agrovoc/c_34017 http://aims.fao.org/aos/agrovoc/c_31727 http://aims.fao.org/aos/agrovoc/c_13251 http://aims.fao.org/aos/agrovoc/c_35130 http://aims.fao.org/aos/agrovoc/c_6242 http://aims.fao.org/aos/agrovoc/c_2604 http://aims.fao.org/aos/agrovoc/c_5731 http://aims.fao.org/aos/agrovoc/c_779 F30 - Génétique et amélioration des plantes H20 - Maladies des plantes Theobroma cacao relation hôte pathogène Moniliophthora péroxydase résistance génétique protéase activité enzymatique résistance aux organismes nuisibles http://aims.fao.org/aos/agrovoc/c_7713 http://aims.fao.org/aos/agrovoc/c_34017 http://aims.fao.org/aos/agrovoc/c_31727 http://aims.fao.org/aos/agrovoc/c_13251 http://aims.fao.org/aos/agrovoc/c_35130 http://aims.fao.org/aos/agrovoc/c_6242 http://aims.fao.org/aos/agrovoc/c_2604 http://aims.fao.org/aos/agrovoc/c_5731 http://aims.fao.org/aos/agrovoc/c_779 Rodrigues Camillo, Luciana Ribeiro Filadelfo, Ciro Monzani, Paulo Sérgio Corrêa, Ronan Xavier Peres Gramacho, Karina Micheli, Fabienne Priminho Pirovani, Carlos Tc-cAPX, a cytosolic ascorbate peroxidase of Theobroma cacao L. engaged in the interaction with Moniliophthora perniciosa, the causing agent of witches' broom disease |
description |
The level of hydrogen peroxide (H2O2) in plants signalizes the induction of several genes, including that of ascorbate peroxidase (APX-EC 1.11.1.11). APX isoenzymes play a central role in the elimination of intracellular H2O2 and contribute to plant responses to diverse stresses. During the infection process in Theobroma cacao by Moniliophthora perniciosa oxidative stress is generated and the APX action recruited from the plant. The present work aimed to characterize the T. cacao APX involved in the molecular interaction of T. cacao-M. perniciosa. The peroxidase activity was analyzed in protein extracts from cocoa plants infected by M. perniciosa and showed the induction of peroxidases like APX in resistant cocoa plants. The cytosolic protein of T. cacao (GenBank: ABR68691.2) was phylogenetically analyzed in relation to other peroxidases from the cocoa genome and eight genes encoding APX proteins with conserved domains were also analyzed. The cDNA from cytosolic APX was cloned in pET28a and the recombinant protein expressed and purified (rTc-cAPX). The secondary structure of the protein was analyzed by Circular Dichroism (CD) displaying high proportion of ?-helices when folded. The enzymatic assay shows stable activity using ascorbate and guaiacol as an electron donor for H2O2 reduction. The pH 7.5 is the optimum for enzyme activity. Chromatographic analysis suggests that rTc-cAPX is a homodimer in solution. Results indicate that the rTc-cAPX is correctly folded, stable and biochemically active. The purified rTc-cAPX presented biotechnological potential and is adequate for future structural and functional studies. |
format |
article |
topic_facet |
F30 - Génétique et amélioration des plantes H20 - Maladies des plantes Theobroma cacao relation hôte pathogène Moniliophthora péroxydase résistance génétique protéase activité enzymatique résistance aux organismes nuisibles http://aims.fao.org/aos/agrovoc/c_7713 http://aims.fao.org/aos/agrovoc/c_34017 http://aims.fao.org/aos/agrovoc/c_31727 http://aims.fao.org/aos/agrovoc/c_13251 http://aims.fao.org/aos/agrovoc/c_35130 http://aims.fao.org/aos/agrovoc/c_6242 http://aims.fao.org/aos/agrovoc/c_2604 http://aims.fao.org/aos/agrovoc/c_5731 http://aims.fao.org/aos/agrovoc/c_779 |
author |
Rodrigues Camillo, Luciana Ribeiro Filadelfo, Ciro Monzani, Paulo Sérgio Corrêa, Ronan Xavier Peres Gramacho, Karina Micheli, Fabienne Priminho Pirovani, Carlos |
author_facet |
Rodrigues Camillo, Luciana Ribeiro Filadelfo, Ciro Monzani, Paulo Sérgio Corrêa, Ronan Xavier Peres Gramacho, Karina Micheli, Fabienne Priminho Pirovani, Carlos |
author_sort |
Rodrigues Camillo, Luciana |
title |
Tc-cAPX, a cytosolic ascorbate peroxidase of Theobroma cacao L. engaged in the interaction with Moniliophthora perniciosa, the causing agent of witches' broom disease |
title_short |
Tc-cAPX, a cytosolic ascorbate peroxidase of Theobroma cacao L. engaged in the interaction with Moniliophthora perniciosa, the causing agent of witches' broom disease |
title_full |
Tc-cAPX, a cytosolic ascorbate peroxidase of Theobroma cacao L. engaged in the interaction with Moniliophthora perniciosa, the causing agent of witches' broom disease |
title_fullStr |
Tc-cAPX, a cytosolic ascorbate peroxidase of Theobroma cacao L. engaged in the interaction with Moniliophthora perniciosa, the causing agent of witches' broom disease |
title_full_unstemmed |
Tc-cAPX, a cytosolic ascorbate peroxidase of Theobroma cacao L. engaged in the interaction with Moniliophthora perniciosa, the causing agent of witches' broom disease |
title_sort |
tc-capx, a cytosolic ascorbate peroxidase of theobroma cacao l. engaged in the interaction with moniliophthora perniciosa, the causing agent of witches' broom disease |
url |
http://agritrop.cirad.fr/571100/ http://agritrop.cirad.fr/571100/1/document_571100.pdf |
work_keys_str_mv |
AT rodriguescamilloluciana tccapxacytosolicascorbateperoxidaseoftheobromacacaolengagedintheinteractionwithmoniliophthoraperniciosathecausingagentofwitchesbroomdisease AT ribeirofiladelfociro tccapxacytosolicascorbateperoxidaseoftheobromacacaolengagedintheinteractionwithmoniliophthoraperniciosathecausingagentofwitchesbroomdisease AT monzanipaulosergio tccapxacytosolicascorbateperoxidaseoftheobromacacaolengagedintheinteractionwithmoniliophthoraperniciosathecausingagentofwitchesbroomdisease AT correaronanxavier tccapxacytosolicascorbateperoxidaseoftheobromacacaolengagedintheinteractionwithmoniliophthoraperniciosathecausingagentofwitchesbroomdisease AT peresgramachokarina tccapxacytosolicascorbateperoxidaseoftheobromacacaolengagedintheinteractionwithmoniliophthoraperniciosathecausingagentofwitchesbroomdisease AT michelifabienne tccapxacytosolicascorbateperoxidaseoftheobromacacaolengagedintheinteractionwithmoniliophthoraperniciosathecausingagentofwitchesbroomdisease AT priminhopirovanicarlos tccapxacytosolicascorbateperoxidaseoftheobromacacaolengagedintheinteractionwithmoniliophthoraperniciosathecausingagentofwitchesbroomdisease |
_version_ |
1792498546385092608 |