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Proteomic analysis of changes in the extracellular matrix of Arabidopsis cell suspension cultures induced by fungal elicitors
Author(s) -
Ndimba Bongani K.,
Chivasa Stephen,
Hamilton John M.,
Simon William J.,
Slabas Antoni R.
Publication year - 2003
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200300413
Subject(s) - elicitor , biology , proteome , biochemistry , xyloglucan , extracellular , receptor tyrosine kinase , microbiology and biotechnology , kinase , cell wall , enzyme
A proteomic approach has been applied to investigate changes in the extracellular matrix of Arabidopsi s thaliana cell suspension cultures following treatments with two fungal pathogen elicitors, chitosan and extracts of Fusarium moniliforme . The oxidative burst and induction of glutathione S ‐transferase were used as markers for induction of the pathogen defence response. Changes in the cell wall and culture filtrate proteome were profiled. Proteins whose abundance changed reproducibly were analysed via matrix assisted laser desorption ionisation‐time of flight (MALDI‐TOF) mass spectrometry (MS). An increase in the level of two classical cell wall proteins (a putative endochitinase and a polygalacturonase inhibiting protein) and two novel proteins (a putative receptor‐like protein kinase and a probable apospory‐associated protein) were seen at 24 hours following elicitation. The level of an unknown protein and a hypothetical protein, which has some homology to serine carboxypeptidases, were decreased at 24 hours post‐elicitation. In the culture filtrate extracts, we identified two pathogen elicitor responsive proteins, a xyloglucan endo‐1,4‐β‐ D glucanases (XEG) and a peroxidase. Using a combination of two‐dimensional polyacrylamide gel electrophoresis, immunoblotting with a phosphotyrosine‐specific antibody, and MALDI‐TOF MS we discovered that spots that represent putative lectin receptor‐like kinase, a putative endochitinase and a XEG possess phosphorylated tyrosine residues. The identification of phosphorylated bona fide cell wall proteins and a putative extracellular receptor‐like kinase with no transmembrane domain implicate the existence of an extracellular phosphorylation network which could be involved in intercellular communication.