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Measurement of carbon flux through the MEP pathway for isoprenoid synthesis by 31 P‐NMR spectroscopy after specific inhibition of 2‐ C ‐methyl‐ d ‐erythritol 2,4‐cyclodiphosphate reductase. Effect of light and temperature
Author(s) -
MONGÉLARD GAËLLE,
SEEMANN MYRIAM,
BOISSON ANNEMARIE,
ROHMER MICHEL,
BLIGNY RICHARD,
RIVASSEAU CORINNE
Publication year - 2011
Publication title -
plant, cell and environment
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.646
H-Index - 200
eISSN - 1365-3040
pISSN - 0140-7791
DOI - 10.1111/j.1365-3040.2011.02322.x
Subject(s) - spinacia , terpenoid , pisum , chemistry , flux (metallurgy) , stereochemistry , mevalonate pathway , substrate (aquarium) , botany , reductase , biochemistry , enzyme , biology , organic chemistry , ecology , chloroplast , gene
The methylerythritol 4‐phosphate (MEP) and the mevalonate pathways are the unique synthesis routes for the precursors of all isoprenoids. An original mean to measure the carbon flux through the MEP pathway in plants is proposed by using cadmium as a total short‐term inhibitor of 2‐ C ‐methyl‐ d ‐erythritol 2,4‐cyclodiphosphate (MEcDP) reductase (GcpE) and measuring the accumulation rate of its substrate MEcDP by 31 P‐NMR spectroscopy. The MEP pathway metabolic flux was determined in spinach ( Spinacia oleracea ), pea ( Pisum sativum ), Oregon grape ( Mahonia aquifolium ) and boxwood ( Buxus sempervirens ) leaves. In spinach, flux values were compared with the synthesis rate of major isoprenoids. The flux increases with light intensity (fourfold in the 200–1200 µ mol m −2 s −1 PPFR range) and temperature (sevenfold in the 25–37 °C range). The relationship with the light and the temperature dependency of isoprenoid production downstream of the MEP pathway is discussed.