title: Involvement of peroxidase activity in developing somatic embryos of Medicago arborea L. Identification of an isozyme peroxidase as biochemical marker of somatic embryogenesis.
abstract: The legume Medicago arborea L. is very interesting as regards the regeneration of marginal arid soils. The problem is that it does not have a good germinative yield. It was therefore decided to regenerate via somatic embryogenesis and find a marker of embryogenic potential. In this study, peroxidase activity was evaluated in non-embryogenic and embryogenic calli from M. arborea L. A decrease in soluble peroxidase activity is observed in its embryonic calli at the time at which the somatic embryos begin to appear. This activity is always lower in embryonic calli than in non-embryonic ones (unlike what happens in the case of wall-bound peroxidases). These results suggest that peroxidases can be considered to be enzymes involved in somatic embryogenesis in M. arborea. In addition, isozyme analyses were carried out on protein extracts using polyacrylamide gel electrophoresis. The band called P5 was detected only in embryogenic cultures at very early stages of development. This band was digested with trypsin and analyzed using linear ion trap (LTQ) mass spectrometer. In P5 isoform a peroxidase-L-ascorbate peroxidase was identified. It can be used as a marker that allows the identification of embryological potential.
authors: Departamento de Fisiología Vegetal, Facultad de Biología, Universidad de Salamanca, Plaza de los Doctores de la Reina s/n, E-37007 Salamanca, Spain.
authors: Piedad
authors: P
authors: Gallego
authors: Departamento de Fisiología Vegetal, Facultades de Biología, Universidad Complutense de Madrid, Madrid, Spain.
authors: Luisa
authors: L
authors: Martin
authors: Departamento de Estadística, Facultad de Economía y Empresa, Universidad de Salamanca, Plaza de los Doctores de la Reina s/n, E-37007 Salamanca, Spain.
authors: Antonio
authors: A
authors: Blazquez
authors: Departamento de Fisiología Vegetal, Facultad de Biología, Universidad de Salamanca, Plaza de los Doctores de la Reina s/n, E-37007 Salamanca, Spain.
authors: Hilario
authors: H
authors: Guerra
authors: Departamento de Fisiología Vegetal, Facultad de Biología, Universidad de Salamanca, Plaza de los Doctores de la Reina s/n, E-37007 Salamanca, Spain. Electronic address: crta@usal.es.
authors: Nieves
authors: N
authors: Villalobos
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D029902:Medicago / Q000196:embryology* / Q000201:enzymology; D009195:Peroxidase / Q000378:metabolism*; D058439:Plant Somatic Embryogenesis Techniques
chemical_list: D015415:Biomarkers; D007527:Isoenzymes; D009195:Peroxidase
keywords: 4-MN; 4-metoxy-α-naphtol; BA; Embryogenic markers; IEF; Isoelectric focusing; Isoenzymes; KIN; LTQ; MALDI-TOF; MS medium supplemented with 2.25mM 2,4-D without cytokinins; MS medium supplemented with 2mgl(−1) of 2,4-D; MS medium supplemented with 9μM 2,4-D and 1μM thidiazuron; MS medium supplemented with 9μM 2,4-D and 9μM benzyladenine; MS medium supplemented with 9μM 2,4-D and 9μM kinetin; MS/MS; MSI; MSI+BA; MSI+KIN; MSI+TDZ; MSII; MSMurashige and Skoog (1962); Medicago arborea L.; Peroxidase activity; Somatic embryogenesis; TDZ; benzyladenine; kinetin; linear ion trap; matrix-assisted laser desorption/ionization time-of-flight; médium; tandem mass spectrometry; thidiazuron