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Comparison of maize brown-midrib isogenic lines by cellular UV-microspectrophotometry and comparative transcript profiling

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Abstract

The molecular mechanisms underlying cell wall digestibility in maize (Zea mays L.) have been studied in three sets of maize brown-midrib isogenic lines in the genetic background of inbreds 1332 (1332 and 1332 bm3), 5361 (5361 and 5361 bm3), and F2 (F2, F2 bm1, F2 bm2, and F2 bm3). Two complementary approaches, SSH (suppression subtractive hybridization) and microarray-based expression profiling, were used to isolate and identify candidate genes in isogenic lines for bm mutants. Metabolic pathway analysis revealed that transcriptional events caused by altering the expression of a single bm gene involve all metabolic and signaling pathways. 53 ESTs were differentially expressed in all three isogenic bm3 comparisons, whereas 32 ESTs were consistently differentially expressed in different bm isogenic lines in F2 background. About 70% ESTs isolated by SSH were not present on the unigene microarray, demonstrating the usefulness of the SSH procedure to identify genes related to cell wall digestibility. Together with lignin analysis by cellular UV-microspectrophotometry, expression profiling in isogenic bm lines proved to be useful to understand alterations at the sub-cellular and molecular level with respect to lignin composition. The down-regulation of COMT affected the expression of CCoAOMT genes and caused a reduced content both of G and S units in bm3 mutants.

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Acknowledgements

We are grateful to Patrick S. Schnable for offering maize unigene-microarrays. We thank Preben B. Holm, Per Gregersen, and Christina Ingvardsen for assistance with microarray experiments in Research Centre Flakkebjerg.

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Authors and Affiliations

  1. Chair of Plant Breeding, Technical University of Munich, Am Hochanger 2, 85350 , Freising, Germany

    Chun Shi, Gerhard Wenzel & Imad Zein

  2. Institute for Wood Biology and Wood Protection, Federal Research Centre for Forestry and Forest Products, Leuschnerstrasse 91, 21031, Hamburg, Germany

    Gerald Koch

  3. KWS Saat AG, Grimsehlstrasse 31, 37555, Einbeck, Germany

    Milena Ouzunova

  4. Department of Genetics and Biotechnology, Research Centre Flakkebjerg, 4200, Slagelse, Denmark

    Thomas Lübberstedt

Authors
  1. Chun Shi
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  2. Gerald Koch
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  3. Milena Ouzunova
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  4. Gerhard Wenzel
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  5. Imad Zein
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  6. Thomas Lübberstedt
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Correspondence to Thomas Lübberstedt.

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Shi, C., Koch, G., Ouzunova, M. et al. Comparison of maize brown-midrib isogenic lines by cellular UV-microspectrophotometry and comparative transcript profiling. Plant Mol Biol 62, 697–714 (2006). https://doi.org/10.1007/s11103-006-9049-3

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  • DOI: https://doi.org/10.1007/s11103-006-9049-3

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