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In Vivo31P and Multilabel13C NMR Measurements for Evaluation of Plant Metabolic Pathways

✍ Scribed by Sushil K. Rijhwani; Chih-Huang Ho; Jacqueline V. Shanks

Publisher
Elsevier Science
Year
1999
Tongue
English
Weight
796 KB
Volume
1
Category
Article
ISSN
1096-7176

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✦ Synopsis

Reliable measurements of intracellular metabolites are useful for effective plant metabolic engineering. This study explored the application of in situ 31 P and 13 C NMR spectroscopy for long-term measurements of intracellular pH and concentrations of several metabolites in glycolysis, glucan synthesis, and central carbon metabolic pathways in plant tissues. An NMR perfusion reactor system was designed to allow Catharanthus roseus hairy root cultures to grow for 3 6 weeks, during which time NMR spectroscopy was performed. Constant cytoplasmic pH (7.40\0.06), observed during the entire experiment, indicated adequate oxygenation. 13 C NMR spectroscopy was performed on hairy root cultures grown in solutions containing 1-13 C-, 2-13 C-, and 3-13 C-labeled glucose in separate experiments and the flow of label was monitored. Activities of pentose phosphate pathways, nonphotosynthetic CO 2 fixation, and glucan synthesis pathways were evident from the experimental results. Scrambling of label in glucans also indicated recycling of triose phosphate and their subsequent conversion to hexose phosphates.

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