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A new method for isolating large quantities of Arabidopsis trichomes for transcriptome, cell wall and other types of analyses

  1. M. David Marks1,*,
  2. Lissete Betancur2,
  3. Edward Gilding1,
  4. Fang Chen3,
  5. Stefan Bauer4,
  6. Jonathan P. Wenger1,
  7. Richard A. Dixon3,
  8. Candace H. Haigler2,5

Article first published online: 4 JUL 2008

DOI: 10.1111/j.1365-313X.2008.03611.x

Issue

The Plant Journal

The Plant Journal

Volume 56, Issue 3, pages 483–492, November 2008

Additional Information

How to Cite

Marks, M. D., Betancur, L., Gilding, E., Chen, F., Bauer, S., Wenger, J. P., Dixon, R. A. and Haigler, C. H. (2008), A new method for isolating large quantities of Arabidopsis trichomes for transcriptome, cell wall and other types of analyses. The Plant Journal, 56: 483–492. doi: 10.1111/j.1365-313X.2008.03611.x

Author Information

  1. 1

    Department of Plant Biology, University of Minnesota, 1445 Gortner Ave., St Paul, MN 55108, USA,

  2. 2

    Department of Plant Biology, North Carolina State University, Box 7612, Raleigh, NC 27695-7612, USA,

  3. 3

    Plant Biology Division, Samuel Roberts Noble Foundation, Ardmore, OK 73401, USA,

  4. 4

    Department of Plant Biology, Carnegie Institution of Washington, Stanford, CA 94305, USA, and

  5. 5

    Department of Crop Science, North Carolina State University, Box 7620, Raleigh, NC 27695-7620, USA

* (fax +1 612 625 1738; e-mail marks004@umn.edu).

Publication History

  1. Issue published online: 22 OCT 2008
  2. Article first published online: 4 JUL 2008
  3. Received 6 February 2008; revised 27 May 2008; accepted 9 June 2008; published online 1 August 2008.

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Keywords:

  • trichome isolation;
  • trichome cell wall;
  • trichome gene expression;
  • lignin;
  • pectin

Summary

A new procedure has been developed for the isolation of wild-type and mutant Arabidopsis trichomes. The isolated trichomes maintained enzymatic activity and were used for DNA, protein, and RNA isolation. The RNA was used to generate probes suitable for Affymetrix analysis. The validity of the Affymetrix results was confirmed by quantitative PCR analysis on a subset of genes that are preferentially expressed in trichomes or leaves. Sufficient quantities of trichomes were isolated to probe the biochemical nature of trichome cell walls. These analyses provide evidence for the presence of lignin in Arabidopsis trichome cell walls. The monosaccharide analysis and positive staining with ruthenium red indicates that the walls also contain a large portion of pectin. The 2.23-fold ratio of pectin-related sugars compared with potential cellulosic glucose suggests that the polysaccharides of the trichome cell walls are more like those of typical primary walls even though the wall becomes quite thick. Overall, these analyses open the door to using the Arabidopsis trichome cell wall as an excellent model to probe various questions concerning plant cell wall biosynthesis.

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