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Gene structure and molecular analysis of the laccase-like multicopper oxidase (LMCO) gene family in Arabidopsis thaliana

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Abstract

Completed genome sequences have made it clear that multicopper oxidases related to laccase are widely distributed as multigene families in higher plants. Laccase-like multicopper oxidase (LMCO) sequences culled from GenBank and the Arabidopsis thaliana genome, as well as those from several newly cloned genes, were used to construct a gene phylogeny that clearly divided plant LMCOs into six distinct classes, at least three of which predate the evolutionary divergence of angiosperms and gymnosperms. Alignments of the predicted amino acid sequences highlighted regions of variable sequence flanked by the highly conserved copper-binding domains that characterize members of this enzyme family. All of the predicted proteins contained apparent signal sequences. The expression of 13 of the 17 LMCO genes in A. thaliana was assessed in different tissues at various stages of development using RT-PCR. A diversity of expression patterns was demonstrated with some genes being expressed in a constitutive fashion, while others were only expressed in specific tissues at a particular stage of development. Only a few of the LMCO genes were expressed in a pattern that could be considered consistent with a major role for these enzymes in lignin deposition. These results are discussed in the context of other potential physiological functions for plant LMCOs, such as iron metabolism and wound healing.

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Abbreviations

BLAST:

Basic local alignment search tool

CTAB:

Cetyl trimethylammonium bromide

LMCO:

Laccase-like multicopper oxidase

MCO:

Multicopper oxidase

MPSS:

Massively parallel signature sequencing

PCR:

Polymerase chain reaction

pI:

Isoelectric point

RACE:

Rapid amplification of cDNA ends

RT-PCR:

Reverse transcript polymerase chain reaction

TBE:

Tris borate EDTA buffer

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Acknowledgements

The authors wish to thank Chieh-Ting Wang for his assistance with tissue collection and RT-PCR assays. This work was supported by and NSF/Alfred P. Sloan Postdoctoral Fellowship (DBI-9803949) to B.C.M and a grant from the U.S. Department of Energy, Energy Biosciences Program (DE-FG02-99ER20336) to J.F.D.D.

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  1. Bonnie C. McCaig

    Present address: Department of Energy-Plant Research Laboratory, Michigan State University, East Lansing, MI, 48824, USA

Authors and Affiliations

  1. Daniel B. Warnell School of Forest Resources, University of Georgia, Athens, GA, 30602, USA

    Bonnie C. McCaig & Jeffrey F. D. Dean

  2. Department of Genetics, University of Georgia, Athens, GA, 30602, USA

    Richard B. Meagher

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  1. Bonnie C. McCaig
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Correspondence to Jeffrey F. D. Dean.

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McCaig, B.C., Meagher, R.B. & Dean, J.F.D. Gene structure and molecular analysis of the laccase-like multicopper oxidase (LMCO) gene family in Arabidopsis thaliana. Planta 221, 619–636 (2005). https://doi.org/10.1007/s00425-004-1472-6

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