Planta

, Volume 219, Issue 4, pp 694–704 | Cite as

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

  • Priya Ranjan
  • Yu-Ying Kao
  • Hongying Jiang
  • Chandrashekhar P. Joshi
  • Scott A. Harding
  • Chung-Jui Tsai
Original Article

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.

Keywords

Expressed sequence tag Phenylpropanoid metabolism Populus Suppression subtractive hybridization Transgenic aspen 

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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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Priya Ranjan
    • 1
  • Yu-Ying Kao
    • 1
    • 2
  • Hongying Jiang
    • 1
  • Chandrashekhar P. Joshi
    • 1
  • Scott A. Harding
    • 1
  • Chung-Jui Tsai
    • 1
  1. 1.Plant Biotechnology Research Center, School of Forest Resources and Environmental ScienceMichigan Technological UniversityHoughtonUSA
  2. 2.Division of Medical Engineering ResearchNational Health Research InstituteTaipeiTaiwan

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