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Identification and isolation of a pectin methylesterase isoform that could be involved in flax cell wall stiffening

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Abstract

Pectin methylesterases (PMEs) are ubiquitous enzymes present in the plant cell wall. They catalyse the demethylesterification of homogalacturonic acid units of pectins, which, in turn, can be associated with different physiological phenomena. In this study, different flax (Linum usitatissimum L.) PME isoforms were observed: neutral (pI 7.0 and 7.5, MW: 110 kDa), basic (pI 8.3 and 8.5, MW: 110 kDa) and very basic (pI>9.5, MW: 38 kDa). In an attempt to identify most of the expressed cell wall LuPME isoforms, polyclonal antibodies were raised against a conserved region of PME. These antibodies allowed the purification of the very basic PME isoform (pI 9.5, MW: 36 kDa) from flax cells, designated LuPME5. This isoform corresponds to the Lupme5 cDNA isolated, at the same time, from flax hypocotyls, by using the RACE–PCR technique. Semi-quantitative PCR experiments showed that the Lupme5 transcript was highly expressed in the hypocotyl zones where elongation is being achieved. Thus, this enzyme may be involved in cell wall stiffening.

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Abbreviations

GST :

Glutathione S-transferase

IEF :

Isoelectric focusing

pI :

Isoelectric point

MALDI–TOF :

Matrix-assisted laser desorption/ionisation–time of flight

PME :

Pectin methylesterase

RACE :

Rapid amplification of cDNA ends

RT–PCR:

Reverse transcription–polymerase chain reaction

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Acknowlegements

S. Al-Qsous and E. Carpentier contributed equally to this work. We acknowledge the région Haute-Normandie (MIIAT-BP; E.C.) and the Jordanian Kingdom (S.A.-Q.) for their financial support.

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Authors and Affiliations

  1. Laboratoire Signaux et régulation chez les végétaux, CNRS UMR 6037, IFRMP 23, Université de Rouen, 76821, Mont-Saint-Aignan Cedex, France

    Suha Al-Qsous, Emilie Carpentier, Dominique Klein-Eude, Carole Burel, Alain Mareck, Hélène Dauchel, Véronique Gomord & Alain Pierre Balangé

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  1. Suha Al-Qsous
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  2. Emilie Carpentier
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  3. Dominique Klein-Eude
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  4. Carole Burel
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  5. Alain Mareck
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  6. Hélène Dauchel
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  7. Véronique Gomord
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  8. Alain Pierre Balangé
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Corresponding author

Correspondence to Alain Pierre Balangé.

Additional information

Accession numbers: Lupme1, AF355056; Lupme3, AF188895; Lupme5, AF355057

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Al-Qsous, S., Carpentier, E., Klein-Eude, D. et al. Identification and isolation of a pectin methylesterase isoform that could be involved in flax cell wall stiffening. Planta 219, 369–378 (2004). https://doi.org/10.1007/s00425-004-1246-1

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  • DOI: https://doi.org/10.1007/s00425-004-1246-1

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