© 2007 Heron Publishing—Victoria, Canada
Identification of genes differentially expressed during adventitious shoot induction in Pinus pinea cotyledons by subtractive hybridization and quantitative PCR
Pablo Alonso (1), Millán Cortizo (1), Francisco R. Cantón (2), Belén Fernández (1), Ana Rodríguez (1), Maria L. Centeno (3), Francisco M. Cánovas (2) and Ricardo J. Ordás (1, 4)
1. Área de Fisiología Vegetal, Departamento BOS, Instituto Universitario de Biotecnología de Asturias, Universidad de Oviedo,
E-33071, Oviedo, Spain / 2. Departamento de Biología Molecular y Bioquímica, Instituto Andaluz de Biotecnología, Unidad Asociada UMA-CSIC, Universidad
de Málaga, E-29071, Málaga, Spain / 3. Departamento de Biología Vegetal, Facultad de Ciencias Biológicas y Ambientales, Universidad de León, E-27071, León, Spain / 4. Corresponding author (rordas@uniovi.es) / Received February 5, 2007; accepted April 23, 2007; published online September 4, 2007
Summary
As part of a study aimed at understanding the physiological and molecular mechanisms involved in adventitious shoot bud formation
in pine cotyledons, we conducted a transcriptome analysis to identify early-induced genes during the first phases of adventitious
caulogenesis in Pinus pinea L. cotyledons cultured in the presence of benzyladenine. A subtractive cDNA library with more than 700 clones was constructed.
Of these clones, 393 were sequenced, analyzed and grouped according to their putative function. Quantitative real-time PCR
analysis was performed to confirm the differential expression of 30 candidate genes. Results are contrasted with available
data for other species.
Keywords:
caulogenesis, conifers, cytokinins, organogenesis, SSH, stone pine.
