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Metabolism and Enzymology

Substrate Utilization by Suspension Cultures and Somatic Embryos of Daucus carota L. Measured by ¹³C NMR

Department of Molecular Physics, Agricultural University Wageningen, DreyenLaan 3, NL 6703 HA Wageningen, The Netherlands, Department of Molecular Biology, Agricultural University Wageningen, DreyenLaan 3, NL 6703 HA Wageningen, The Netherlands

The uptake and utilization of sucrose by embryogenic suspension cultures of carrot (Daucus carota L.) growing in the presence of 2,4-D and by somatic embryos derived from these cultures was monitored using ¹³C nuclear magnetic resonance. The exogeneously supplied sucrose was completely hydrolyzed before cell entry; glucose was taken up preferentially when the cells were cultured in the presence of 2,4-D, while glucose and fructose were utilized at similar rates by somatic embryos in the absence of 2,4-D. Both suspension cells and somatic embryos accumulated high intracellular levels predominantly of glucose and sucrose, the latter being resynthesized intracellularly from the constitutive hexoses. Initially, fructose was converted mainly into glucose and sucrose rather than being catabolized directly through glycolysis or the pentose phosphate pathway. Carbohydrate supply that exceeded cellular demand resulted in intracellular accumulation of mono- or disaccharides. The capacity of cultured carrot cells to produce somatic embryos appeared to be positively correlated with high intracellular levels of glucose.

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