Abstract
Determination of soluble sugars is basic for the study of carbon metabolism in plants. Soluble sugar quantitation can be achieved by enzymatic methods implying different coupled reactions. Here we describe a simple method that allows rapid determination of the most abundant soluble sugars (glucose, fructose and sucrose) in Arabidopsis leaves by anion exchange chromatography. We have applied this method to study the levels of soluble sugars during the photoperiodic transition to flowering (Ortiz-Marchena et al., 2014).
Keywords: Sugar determination, Soluble sugars, Arabidopsis, HPLC
Materials and Reagents
Equipment
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Procedure
Representative data
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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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Hi, HayatAs said in the protocol, first you inject into the column a mixture of sugars (threalose, galactose, glucose, fructose and sucrose) at different known concentrations (1, 0.5, 0.1, 0.05 and 0.01 mM; you can see the chromatogram of 0.5 mM mixture in figure 1A). For each sugar, you record the peak area at each concentration and with these data you can generate a calibration curve, that is, the mathematical function that relates both variables (peak area (x) and concentration (y). Then, once you inject your sample and record the area of the peak of your sugar(s) of interest, all you have to do is to use that value as “x” in the calibration curve you have obtained.I hope I understood your question properly. If not, please feel free to contact me.