Abstract
Starch constitutes the most important carbon reserve in plants and is composed of branched amylopectin and linear amylose. The latter is synthesized exclusively by the Granule-Bound Starch Synthase (GBSS, EC 2.4.1.21). Here we report a readily reproducible, specific and highly sensitive protocol, which includes the isolation of intact starch granules from Arabidopsis thaliana leaves and the subsequent determination of GBSS activity. We have applied this method to study GBSS activity in diurnal cycles in vegetative growth and during the photoperiodic transition to flowering in Arabidopsis (Tenorio et al., 2003; Ortiz-Marchena et al., 2014).
Keywords: Starch granule, Starch purification, GBSS activity, Granule-bound starch synthase
Materials and Reagents
Equipment
Procedure
Representative data
Notes
Recipes
Note: The given volumes are sufficient for 50 reactions.
Acknowledgments
This work was performed with funding from projects CSD2007-00057, BIO2008-02292, and BIO2011-28847-C02-00 (Spanish Ministry of Economy and Competitiveness, MINECO) and Excellence projects P06-CVI-01450 and P08-AGR-03582 (Junta de Andalucía) partially supported by FEDER funding to F. V. and J. M. R. We also acknowledge the TRANSPLANTA consortium Project CONSOLIDER 28317 (MINECO).
References
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