The induction of differentially expressed proteins of Xylella fastidiosa with citrus extract

An in vitro system was developed to induce and identify Xylella fastidiosa proteins that were differentially expressed in the presence of callus-derived extracts from its host, the citrus cultivar Pêra. To optimize the induction, we first developed a single culture medium for the growth of both, host and bacteria. This medium, CPXPm7, which mimics the citrus xylem sap, showed that X. fastidiosa at 72 h post-incubation had 10(8) colony forming units mL-1, while Pêra cells had the highest fresh weight content (0.79 g). After testing various methods of co-cultivation of the bacteria and host callus grown in this single medium, the best induction procedure was to grow X. fastidiosa in a solid medium amended with an extract of Pêra callus grown in CPXPm7. Analysis, by two-dimensional electrophoresis, of the X. fastidiosa proteins (120 µg of total proteins) grown in the presence of Pêra callus extract revealed 414 differentially expressed protein spots when compared to the protein profile obtained in the absence of the extract. The system developed in this study improves the induction and analysis of differentially expressed proteins of X. fastidiosa, which may be involved in pathogenicity.

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Main Authors: Bellato,Cláudia de M., Garcia,Andréia K.M., Mestrinelli,Fabiana, Tsai,Siu M., Machado,Marcos A., Meinhardt,Lyndel W.
Format: Digital revista
Language:English
Published: Sociedade Brasileira de Microbiologia 2004
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822004000200012
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spelling oai:scielo:S1517-838220040002000122005-11-25The induction of differentially expressed proteins of Xylella fastidiosa with citrus extractBellato,Cláudia de M.Garcia,Andréia K.M.Mestrinelli,FabianaTsai,Siu M.Machado,Marcos A.Meinhardt,Lyndel W. 2DE callus pathogenicity protein Xylella-citrus interaction An in vitro system was developed to induce and identify Xylella fastidiosa proteins that were differentially expressed in the presence of callus-derived extracts from its host, the citrus cultivar Pêra. To optimize the induction, we first developed a single culture medium for the growth of both, host and bacteria. This medium, CPXPm7, which mimics the citrus xylem sap, showed that X. fastidiosa at 72 h post-incubation had 10(8) colony forming units mL-1, while Pêra cells had the highest fresh weight content (0.79 g). After testing various methods of co-cultivation of the bacteria and host callus grown in this single medium, the best induction procedure was to grow X. fastidiosa in a solid medium amended with an extract of Pêra callus grown in CPXPm7. Analysis, by two-dimensional electrophoresis, of the X. fastidiosa proteins (120 µg of total proteins) grown in the presence of Pêra callus extract revealed 414 differentially expressed protein spots when compared to the protein profile obtained in the absence of the extract. The system developed in this study improves the induction and analysis of differentially expressed proteins of X. fastidiosa, which may be involved in pathogenicity.info:eu-repo/semantics/openAccessSociedade Brasileira de MicrobiologiaBrazilian Journal of Microbiology v.35 n.3 20042004-09-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822004000200012en10.1590/S1517-83822004000200012
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countrycode BR
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libraryname SciELO
language English
format Digital
author Bellato,Cláudia de M.
Garcia,Andréia K.M.
Mestrinelli,Fabiana
Tsai,Siu M.
Machado,Marcos A.
Meinhardt,Lyndel W.
spellingShingle Bellato,Cláudia de M.
Garcia,Andréia K.M.
Mestrinelli,Fabiana
Tsai,Siu M.
Machado,Marcos A.
Meinhardt,Lyndel W.
The induction of differentially expressed proteins of Xylella fastidiosa with citrus extract
author_facet Bellato,Cláudia de M.
Garcia,Andréia K.M.
Mestrinelli,Fabiana
Tsai,Siu M.
Machado,Marcos A.
Meinhardt,Lyndel W.
author_sort Bellato,Cláudia de M.
title The induction of differentially expressed proteins of Xylella fastidiosa with citrus extract
title_short The induction of differentially expressed proteins of Xylella fastidiosa with citrus extract
title_full The induction of differentially expressed proteins of Xylella fastidiosa with citrus extract
title_fullStr The induction of differentially expressed proteins of Xylella fastidiosa with citrus extract
title_full_unstemmed The induction of differentially expressed proteins of Xylella fastidiosa with citrus extract
title_sort induction of differentially expressed proteins of xylella fastidiosa with citrus extract
description An in vitro system was developed to induce and identify Xylella fastidiosa proteins that were differentially expressed in the presence of callus-derived extracts from its host, the citrus cultivar Pêra. To optimize the induction, we first developed a single culture medium for the growth of both, host and bacteria. This medium, CPXPm7, which mimics the citrus xylem sap, showed that X. fastidiosa at 72 h post-incubation had 10(8) colony forming units mL-1, while Pêra cells had the highest fresh weight content (0.79 g). After testing various methods of co-cultivation of the bacteria and host callus grown in this single medium, the best induction procedure was to grow X. fastidiosa in a solid medium amended with an extract of Pêra callus grown in CPXPm7. Analysis, by two-dimensional electrophoresis, of the X. fastidiosa proteins (120 µg of total proteins) grown in the presence of Pêra callus extract revealed 414 differentially expressed protein spots when compared to the protein profile obtained in the absence of the extract. The system developed in this study improves the induction and analysis of differentially expressed proteins of X. fastidiosa, which may be involved in pathogenicity.
publisher Sociedade Brasileira de Microbiologia
publishDate 2004
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822004000200012
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