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Identification of genes regulated by ammonium availability in the roots of maritime pine trees

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Abstract

Conifers have a preference for ammonium over nitrate as the main inorganic nitrogen source. However, it is unknown how changes in nitrogen nutrition may affect transcription profiles. In this study, microarray analysis and suppressive subtraction hybridization were used to identify differentially expressed genes in the roots of maritime pine exposed to changes in ammonium availability. A total of 225 unigenes that were differentially regulated by changes in ammonium nutrition were identified. Most of the unigenes were classified into seven functional categories by comparison with sequences deposited in the databases. A significant proportion of these genes were encoded for ammonium-regulated proteins of unknown functions. The differential expression of selected candidate genes was further validated in plants subjected to ammonium excess/deficiency. The transcript levels of representative genes were compared in maritime pine roots, 1, 15 and 35 days after nutritional treatments. Gene expression patterns suggest the existence of potential links between ammonium-responsive genes and genes involved in amino acid metabolism, particularly in asparagine biosynthesis and utilization. Functional analyses and exploration of the natural variability in maritime pine populations for a number of relevant genes are underway.

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Acknowledgments

This work was supported by grants from the Spanish Ministry of Science and Innovation (BIO2006-06216 and BIO2009-07940) and by Junta de Andalucía (AGR-663 and funds to BIO-114 research group). JC was supported by a predoctoral fellowship from Junta de Andalucía.

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Authors and Affiliations

  1. Departamento de Biología Molecular y Bioquímica, Instituto Andaluz de Biotecnología, Unidad Asociada UMA-CSIC, Universidad de Málaga, Campus Universitario de Teatinos, 29071, Málaga, Spain

    Javier Canales, Arantxa Flores-Monterrosso, Marina Rueda-López, Concepción Avila & Francisco M. Cánovas

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  1. Javier Canales
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  2. Arantxa Flores-Monterrosso
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  3. Marina Rueda-López
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  4. Concepción Avila
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  5. Francisco M. Cánovas
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Corresponding author

Correspondence to Francisco M. Cánovas.

Electronic Supplementary material

Table 1

Differentially expressed genes identified by microarray analysis microarray (XLS 24 kb)

Table 2

Genes with putative functions identified in the SSH libraries (XLS 151 kb)

Table 3

Sequences of oligonucleotides used for qPCR analysis of gene expression (XLS 23 kb)

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Canales, J., Flores-Monterrosso, A., Rueda-López, M. et al. Identification of genes regulated by ammonium availability in the roots of maritime pine trees. Amino Acids 39, 991–1001 (2010). https://doi.org/10.1007/s00726-010-0483-9

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  • DOI: https://doi.org/10.1007/s00726-010-0483-9

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