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Immunolocalization of two ligninO-methyltransferases in stems of alfalfa (Medicago sativa L.)

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Summary

Caffeic acid 3-O-methyltransferase (COMT) and caffeoyl CoA 3-O-methyltransferase (CCOMT) catalyze parallel reactions that are believed to be involved in the biosynthesis of lignin monomers. Antisera specific for alfalfa (Medicago sativa L.) COMT or CCOMT were raised against the enzymes expressed inEscherichia coli, and were used for immunolocalization studies in lignifying alfalfa stem tissue. Both COMT and CCOMT were localized to xylem parenchyma cells, as assessed by light microscopy and immunocytochemistry. Electron microscopy revealed that both enzymes were located in the cytoplasm of xylem parenchyma cells, and to a lesser extent, in the cytoplasm of phloem cells. There was no significant difference in the localization pattern of COMT and CCOMT, suggesting that the two enzymes may be part of a metabolic grid leading to production of lignin monomers in lignifying tissue of mature alfalfa stem internodes.

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Abbreviations

BSA:

bovine serum albumin

BSE:

backscattered electron

CCOMT:

caffeoyl CoA 3-O-methyltransferase

COMT:

caffeic acid 3-O-methyltransferase

EM:

electron microscope

OMT:

O-methyltransferase

PAGE:

polyacrylamide gel electrophoresis

PBS:

phosphate-buffered saline

PBST:

phosphate-buffered saline containing Tween 20

SDS:

sodium dodecyl sulfate

SE:

secondary electron

SEM:

scanning electron microscope

TBST:

Tris-buffered saline containing Tween 20

TEM:

transmission electron microscope

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Authors and Affiliations

  1. Department of Botany and Microbiology, University of Oklahoma, Norman, Oklahoma

    Rossio Kersey & Karel R. Schubert

  2. Plant Biology Division, The Samuel Roberts Noble Foundation, 2510, Sam Noble Parkway, 73401, Ardmore, OK, USA

    Kentaro Inoue & Richard A. Dixon

Authors
  1. Rossio Kersey
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  2. Kentaro Inoue
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  3. Karel R. Schubert
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  4. Richard A. Dixon
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Correspondence to Richard A. Dixon.

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Kersey, R., Inoue, K., Schubert, K.R. et al. Immunolocalization of two ligninO-methyltransferases in stems of alfalfa (Medicago sativa L.). Protoplasma 209, 46–57 (1999). https://doi.org/10.1007/BF01415700

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  • DOI: https://doi.org/10.1007/BF01415700

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