Plant Molecular Biology

, Volume 61, Issue 1–2, pp 329–344 | Cite as

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

  • Ying Jia
  • James V. Anderson
  • David P. Horvath
  • Yong-Qiang Gu
  • Rodney G. Lym
  • Wun S. Chao


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.


dormancy leafy spurge subtractive hybridization 


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  1. Anderson, J.V., Chao, W.S., Horvath, D.P. 2001A current review on the regulation of dormancy in vegetative budsWeed Sci.49581589Google Scholar
  2. Anderson, J.V., Horvath, D.P. 2001Random sequencing of cDNAs and identification of mRNAsWeed Sci.50227231Google Scholar
  3. Anderson, J.V., Gesch, R.W., Jia, Y., Chao, W.S., Horvath, D.P. 2005Seasonal shifts in dormancy status, carbohydrate metabolism, and related gene expression in crown buds of leafy spurgePlant Cell Environ.2815671578CrossRefGoogle Scholar
  4. Bak, S., Tax, F.E., Feldmann, K.A., Galbraith, D.W., Feyereisen, R. 2001CYP83B1, a cytochrome P450 at the metabolic branch point in auxin and indole glucosinolate biosynthesis in Arabidopsis Plant Cell13101111PubMedCrossRefGoogle Scholar
  5. Bassani, M., Neumann, P.M., Gepstein, S. 2004Differential expression profiles of growth-related genes in the elongation zone of maize primary rootsPlant Mol. Biol.56367380PubMedCrossRefGoogle Scholar
  6. CAB International 2004. Euphorbia esula [original text by W. Chao and J.V. Anderson]. In: Crop Protection Compendium, 2004 edition. CAB International, Wallingford, UK [CD-ROM].Google Scholar
  7. Chang, S., Puryear, J., Cairney, J. 1993A simple and efficient method for isolating RNA from pine treesPlant Mol. Biol. Rep.11113116Google Scholar
  8. Chao, W.S., Serpe, M.D., Anderson, J.V., Gesch, R.W. and Horvath, D.P., 2006. Sugars, hormones, and environment affect the dormancy status in underground adventitious buds of leafy spurge (Euphorbia esula). Weed Sci. 54: 59–68.Google Scholar
  9. Coupland, R.T., Selleck, G.W., Alex, J.F. 1955Distribution of vegetative buds on the underground parts of leafy spurge (Euphorbia esula L.)Can. J. Agric. Sci.35161167Google Scholar
  10. Diatchenko, L., Lau, Y.F., Campbell, A.P., Chenchik, A., Moqadam, F., Huang, B., Lukyanov, S., Lukyanov, K., Gurskaya, N., Sverdlov, E.D., Siebert, P.D. 1996Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and librariesProc. Natl. Acad. Sci. USA9360256030PubMedCrossRefGoogle Scholar
  11. Footitt, S., Slocombe, S.P., Larner, V., Kurup, S., Wu, Y., Larson, T., Graham, I., Baker, A., Holdsworth, M. 2002Control of germination and lipid mobilization by COMATOSE, the Arabidopsis homologue of human ALDPEMBO J.2129122922PubMedCrossRefGoogle Scholar
  12. Frewen, B.E., Chen, T.H.H., Howe, G.T., Davis, J., Rohde, A., Boerjan, W., Bradshaw, H.D.,Jr. 2000Quantitative trait loci and candidate gene mapping of bud set and bud flush in populusGenetics154837845PubMedGoogle Scholar
  13. Freyre, R., Warnke, S., Sosinski, B., Souches, D.S. 1994Quantitative trait locus analysis of tuber dormancy in diploid potato (Solanum spp)Theor. Appl. Genet.89474480CrossRefGoogle Scholar
  14. Gardeal, A.A., Moreno, Y.M., Azarenko, A.N., Lombard, P.B., Daley, L.S., Criddle, R.S. 1994Changes in metabolic properties of grape buds during developmentJ. Am. Soc. Hort. Sci.119756760Google Scholar
  15. Harvey, S.J., Nowierski, R.M. 1988Release of postsenescent dormancy in leafy spurge (Euphorbia esula L.) by chillingWeed Sci.36784786Google Scholar
  16. Horvath, D.P. 1998The role of specific plant organs and polar auxin transport in correlative inhibition of leafy spurge (Euphorbia esula L.) root budsCan. J. Bot.7612271232CrossRefGoogle Scholar
  17. Horvath, D.P. 1999Role of mature leaves in inhibition of root bud growth in Euphorbia esula LWeed Sci.47544550Google Scholar
  18. Horvath, D.P., Anderson, J.V. 2002A molecular approach to understanding root bud dormancy in leafy spurgeWeed Sci.50227231Google Scholar
  19. Horvath, D.P., Chao, W.S., Anderson, J.V. 2002Molecular analysis of signals controlling dormancy and growth in underground adventitious buds of leafy spurge (Euphorbia esula L.)Plant Physiol.12814391446PubMedCrossRefGoogle Scholar
  20. Horvath, D.P., Anderson, J.V., Chao, W.S., Foley, M.E. 2003Knowing when to grow: signals regulating bud dormancyTrends Plant Sci.8534540PubMedCrossRefGoogle Scholar
  21. Horvath, D.P., Anderson, J.V., Jia, Y., Chao, W.S. 2005Cloning, characterization and expression of growth regulator CYCLIN D3–2 in Leafy Spurge (Euphorbia esula)Weed Sci.53431437Google Scholar
  22. Lazo, G.R., Lui, N., Gu, Y.Q., Kong, X., Coleman-Derr, D., Anderson, O.D. 2005Hybsweeper: a resource for detecting high-density plate gridding coordinatesBioTechniques39320324PubMedCrossRefGoogle Scholar
  23. McIntyre, G.I. 1972Developmental studies on Euphorbia esula L. The influence of the nitrogen supply on the correlative inhibition of root bud activityCan. J. Bot.50949956Google Scholar
  24. Nissen, S.J., Foley, M.E. 1987aCorrelative inhibition and dormancy in root buds of leafy spurge (Euphorbia esula L.)Weed Sci.35155159Google Scholar
  25. Nissen, S.J., Foley, M.E. 1987bEuphorbia esula L. root and root bud indole-3-acetic acid levels at three phenologic stagesPlant Physiol.84287290CrossRefGoogle Scholar
  26. Pariasca, J.A.T., Sunaga, A., Miyazaki, T., Hisaka, H., Sonoda, M., Nakagawa, H., Sato, T. 2001Cloning of cDNAs encoding senescence-associated genes, Acc synthase and Acc oxidase from stored snow pea pods (Pisum sativum L. var Saccharatum) and their expression during pod storagePostharvest Biol. Technol.22239247CrossRefGoogle Scholar
  27. Paterson, A.H., Scherz, K.F., Lin, Y.R., Liu, S.C., Chang, Y.L. 1995The weediness of wild plants: molecular analysis of genes influencing dispersal and persistence of johnsongrass, Sorghum halepense (L.) PersProc. Natl. Acad. Sci. USA9261276131PubMedCrossRefGoogle Scholar
  28. Sambrook, J., Fritsch, E.F., Maniatis, T. 1989Molecular Cloning – A Laboratory ManualCold Spring Harbor Laboratory PressCold Spring Harbor, NYGoogle Scholar
  29. Sebastiani, P., Gussoni, E., Kohane, I.S., Ramoni, M.F. 2003Statistical challenges in functional genomicsStatistical Sci.183370CrossRefGoogle Scholar
  30. Šimko, I., McMurry, S., Yang, H.M., Manschot, A., Davies, P.J., Ewing, E.E. 1997Evidence from polygene mapping for a causal relationship between potato tuber dormancy and abscisic acid contentPlant Physiol.11514531459PubMedGoogle Scholar
  31. Sugiura, M., Sakaki, T., Yabusaki, Y., Ohkawa, H. 1996Cloning and expression in Escherichia coli and Saccharomyces cerevisiae of a novel tobacco cytochrome P-450-like cDNABiochim. Biophys. Acta1308231240PubMedGoogle Scholar
  32. Berg, J.H., Ewing, E.E., Plaisted, R.L., McMurry, S., Bonierbale, M.W. 1996QTL analysis of potato tuber dormancyTheor. Appl. Genet.93317324CrossRefGoogle Scholar
  33. Zheng, J., Zhao, J., Tao, Y., Wang, J., Liu, Y., Fu, J., Jin, Y., Gao, P., Zhang, J., Bai, Y., Wang, G. 2004Isolation and analysis of water stress induced genes in maize seedlings by subtractive PCR and cDNA macroarrayPlant Mol. Biol.55807823PubMedGoogle Scholar

Copyright information

© Springer 2006

Authors and Affiliations

  • Ying Jia
    • 1
  • James V. Anderson
    • 2
  • David P. Horvath
    • 2
  • Yong-Qiang Gu
    • 3
  • Rodney G. Lym
    • 1
  • Wun S. Chao
    • 2
  1. 1.Department of Plant SciencesNorth Dakota State UniversityFargoUSA
  2. 2.Plant Science ResearchUSDA-Agricultural Research ServiceFargoUSA
  3. 3.Western Regional Research CenterUSDA-ARSAlbanyUSA

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