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Subtractive cDNA Libraries Identify Differentially Expressed Genes in Dormant and Growing Buds of Leafy Spurge (Euphorbia esula)

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Abstract

Two subtractive cDNA libraries were developed to study genes associated with bud dormancy (reverse library) and initiation of shoot growth (forward library) in leafy spurge. To identify unique sequences represented in each library, 15744 clones were screened to reduce the level of redundancy within both libraries. A total of 516 unique sequences were obtained from 2304 minimally redundant clones. Radioactive probes developed from RNAs extracted from crown buds of either intact (para-dormant control) or a series of growth-induced (2 h, 2, and 4 d after decapitation) plants were used to identify differentially expressed genes by macroarray analysis. Semi-quantitative RT-PCR was used to confirm results obtained by macroarray analysis and to determine the expression profiles for other transcripts identified within the subtractive libraries. Selected clones were also used to examine gene expression in crown buds after growth induction and/or during normal seasonal growth. In this study, four distinct patterns of gene expression were observed during the transition from para-dormancy to growth-induction. Many of the differentially regulated genes identified have unknown or hypothetical functions while others are known to play important roles in molecular functions. Gene ontology analysis identified a greater proportion of genes involved with catalytic activity in the forward library while the reverse library had a greater proportion of genes involved in DNA/RNA binding.

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Authors and Affiliations

  1. Department of Plant Sciences, North Dakota State University, Fargo, ND, 58105, USA

    Ying Jia & Rodney G. Lym

  2. Plant Science Research, USDA-Agricultural Research Service, 1605 Albrecht Blvd., Fargo, ND , 58105-5674, USA

    James V. Anderson, David P. Horvath & Wun S. Chao

  3. Western Regional Research Center, USDA-ARS, Albany, CA, 94710, USA

    Yong-Qiang Gu

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  1. Ying Jia
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  2. James V. Anderson
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  3. David P. Horvath
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  4. Yong-Qiang Gu
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  5. Rodney G. Lym
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  6. Wun S. Chao
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Correspondence to Wun S. Chao.

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Jia, Y., Anderson, J.V., Horvath, D.P. et al. Subtractive cDNA Libraries Identify Differentially Expressed Genes in Dormant and Growing Buds of Leafy Spurge (Euphorbia esula). Plant Mol Biol 61, 329–344 (2006). https://doi.org/10.1007/s11103-006-0015-x

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