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J. Biol. Chem., Vol. 262, Issue 11, 5000-5007, Apr, 1987
C Roby, JB Martin, R Bligny and R Douce
An experimental arrangement was described that enables nuclear magnetic
resonance spectra of compressed plant cells to be recorded while
circulating a medium through the sample. The system provided a convenient
arrangement for monitoring by 31P NMR the behavior of plant cells over a
long period of time under different conditions such as sucrose starvation.
Perfusion of compressed sycamore cells with sucrose- free culture medium
triggered a progressive decrease in the glucose 6-P and
uridine-5'-diphosphate-alpha-D-glucose resonances over 30 h. When almost
all the intracellular carbohydrate pool had disappeared the nucleotide
triphosphate resonances decline progressively. These changes were
accompanied by a Pi accumulation in the vacuole and a phosphorylcholine
(P-choline) accumulation in the cytoplasm. The very long lag phase observed
for ATP and P-choline evolution was comparable with that observed for the
progressive intracellular digestion of cytoplasmic constituents (Journet,
E., Bligny, R. and Douce, R. (1986) J. Biol. Chem. 261, 3193-3199).
Addition of sucrose in the circulating system after a long period of
sucrose starvation led to a disappearance of the cytoplasmic Pi resonance
and a marked increase in that of glucose 6-P. Under these conditions the
vacuolar Pi pool did not fluctuate to buffer the Pi in the cytoplasm. The
results suggest that Pi which has been sequestered in the vacuole during
the course of sucrose starvation is not restored to the cytoplasm for rapid
metabolic processes. Furthermore, the presence of P-choline in plant cells
in large excess should be considered as a good marker of membrane
utilization after a long period of sucrose starvation and is very likely
related to stress.
Biochemical changes during sucrose deprivation in higher plant cells. Phosphorus-31 nuclear magnetic resonance studies
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