Observations on the phosphate status and intracellular pH of intact cells, protoplasts and chloroplasts from photosynthetic tissue using phosphorus-31 nuclear magnetic resonance

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Abstract
Individual pools of intracellular inorganic phosphate (Pi) can be observed in the dark in intact cells, protoplasts and chloroplasts from photosynthetic tissue by using 31P nuclear magnetic resonance (n.m.r.). Estimates for the pH of vacuolar and extravacuolar compartments are reported although it is shown that intracellular pH is determined by the pH of the suspending medium. Mannose treatment of asparagus (Asparagus officinalis) cells and spinach (Spinacia oleracea) protoplasts results in the inhibition of photosynthesis. The mechanism of mannose phosphate sequestration of free Pi is supported by the 31P n.m.r. spectra of mannose-treated tissue. There is a fundamental difference in 31 P n.m.r. spectra of mannose-treated spinach protoplasts and asparagus cells, reflecting a difference in the availability of vacuolar Pi for cellular metabolism in these species. The 31P n.m.r. spectrum of intact spinach chloroplasts is reported.