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Suppression subtractive hybridization-mediated transcriptome analysis from multiple tissues of aspen (Populus tremuloides) altered in phenylpropanoid metabolism

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Abstract

A PCR-based suppression subtractive hybridization (SSH) technique was used to identify differentially expressed genes in developing tissues of control and transgenic aspen (Populus tremuloides Michx.) with down-regulated 4CL1 (4-coumarate:coenzyme A ligase) expression and enhanced growth. A total of 11,308 expressed sequence tags (ESTs) representing 5,028 non-redundant transcripts encoding 4,224 unique proteins was obtained from shoot apex, young stem, young leaf and root tip SSH libraries. Putative functions can be assigned to 60% of these transcripts. Approximately 14% of the ESTs are not represented among the 111,000 entries already present in Populus EST databases. In general, ESTs of the metabolism class occurred at a higher frequency in control- than transgenic-enriched libraries of all tissues, whereas protein synthesis and protein fate ESTs were over-represented in meristematic tissues of transgenics where 4CL1 was relatively strongly suppressed. Among all tissues, leaves yielded the highest percentage of ESTs with either unknown protein function or insignificant similarity to other protein/DNA/EST sequences in existing databases. Of particular interest was a large number of ESTs (16%) associated with signal transduction in transgenic leaves. Among these were several leucine-rich-repeat receptor-like protein kinases with markedly elevated expression in transgenic leaves. We also identified homologs of transposable elements that were up-regulated in transgenic tissues, providing the first experimental data for active expression of DNA mobile elements in long-lived tree species.

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Fig. 1a, b

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Abbreviations

CA, CL, CR, CS:

Control apices, leaves, stems, roots

4CL :

4-Coumarate:coenzyme A ligase

EST:

Expressed sequence tag

Lhcb2 :

Light-harvesting chlorophyll a/b-binding protein, type II

LRR-RLK:

Leucine-rich-repeat receptor-like serine/threonine protein kinase

SSH:

Suppression subtractive hybridization

TA, TL, TR, TS:

Transgenic apices, leaves, stems, roots

TE:

Transposable element

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Acknowledgements

We thank Prof. Vincent Chiang (North Carolina State University) for providing the transgenic aspen lines. This work was supported by the Michigan Life Sciences Corridor Funds of the Michigan Economic Development Corporation (grant no. 085P1000477).

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Author notes

  1. Yu-Ying Kao

    Present address: Division of Medical Engineering Research, National Health Research Institute, Taipei, Taiwan

Authors and Affiliations

  1. Plant Biotechnology Research Center, School of Forest Resources and Environmental Science, Michigan Technological University, 1400 Townsend Drive, Houghton, MI 49931, USA

    Priya Ranjan, Yu-Ying Kao, Hongying Jiang, Chandrashekhar P. Joshi, Scott A. Harding & Chung-Jui Tsai

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  1. Priya Ranjan
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Correspondence to Chung-Jui Tsai.

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Ranjan, P., Kao, YY., Jiang, H. et al. Suppression subtractive hybridization-mediated transcriptome analysis from multiple tissues of aspen (Populus tremuloides) altered in phenylpropanoid metabolism. Planta 219, 694–704 (2004). https://doi.org/10.1007/s00425-004-1291-9

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