Abstract
Glutamate decarboxylase (GAD, EC 4.1.1.15) is a key enzyme in the synthesis of γ-aminobutyric acid (GABA) in higher plants. A complete cDNA encoding glutamate decarboxylase (GAD, EC 4.1.1.15) was characterized from Pinus pinaster Ait, and its expression pattern was studied to gain insight into the role of GAD in the differentiation of the vascular system. Pine GAD contained a C-terminal region with conserved residues and a predicted secondary structure similar to the calmodulin (CaM)-binding domains of angiosperm GADs. The enzyme was able to bind to a bovine CaM-agarose column and GAD activity was higher at acidic pH, suggesting that the pine GAD can be regulated in vivo by Ca2+/CaM and pH. A polyclonal antiserum was prepared against the pine protein. GAD expression was studied at activity, protein, and mRNA level and was compared with the expression of other genes during the differentiation of the hypocotyl and induction of reaction wood. In seedling organs, GABA levels closely matched GAD expression, with high levels in the root and during lignification of the hypocotyl. GAD expression was also induced in response to the production of compression wood and its expression matched the pattern of other genes involved in ethylene and 2-oxoglutarate synthesis. The results suggest of a role of GAD in hypocotyl and stem development in pine.
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Abbreviations
- ACO:
-
1-Aminocyclopropane-1-carboxylic acid oxidase
- ACS:
-
1-Aminocyclopropane-1-carboxylic acid synthase
- APX:
-
Ascorbate peroxidase
- AOX:
-
Ascorbate oxidase
- CaM:
-
Calmodulin
- GAD:
-
Glutamate decarboxylase
- LAC:
-
Laccase
- NADP+-IDH:
-
NADP+-isocitrate dehydrogenase
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Acknowledgments
We want to thank Fernando Nicolás de la Torre Fazio for his help in the production of polyclonal antibodies. This work was supported by the grants BIO2003-04590, and AGL2009-11404 to F.G, and BIO2006-06216 to F.M.C. from the Spanish Ministry of Science and Innovation, which were partially financed by FEDER funds from the European Union. J.J.M.-R. and M.B.P. were recipients of a FPU and a UAs-CSIC fellowship from the Spanish Ministry of Education and Science, respectively.
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Molina-Rueda, J.J., Pascual, M.B., Cánovas, F.M. et al. Characterization and developmental expression of a glutamate decarboxylase from maritime pine. Planta 232, 1471–1483 (2010). https://doi.org/10.1007/s00425-010-1268-9
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DOI: https://doi.org/10.1007/s00425-010-1268-9