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Plant Physiology 75:1001-1008 (1984)
© 1984 American Society of Plant Biologists

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Articles

Factors Permitting Prolonged Translation by Isolated Pea Chloroplasts 1

Helen T. Nivison2 and Andre T. Jagendorf

Plant Biology Section, Cornell University, Ithaca, New York 14853

The following parameters were found to prolong the time-course of translation by isolated pea (Pisum sativum, cv Progress No. 9) chloroplasts: addition of other amino acids (an effect synergistic with sufficient free Mg2+), use of lower light intensities, and additions of inorganic phosphate and ATP. In a chloroplast system which includes these parameters, active translation usually extends to almost an hour. The total amount of leucine incorporated is routinely 60 to 100 nanomoles/milligram chlorophyll and often 200 nanomoles/milligram chlorophyll. Accurate estimation of the amount of amino acid incorporated depends on supplying the labeled amino acid at a concentration sufficient to overcome isotope dilution effects from endogenous pools. Approximately 39 thylakoid and 60 stroma polypeptides were visible on autoradiographs after labeling with [35S]methionine. Label in a few of the polypeptide bands was increased or decreased by specific changes in the reaction conditions. Due to the long period of activity and the large number of labeled products, this chloroplast system should be useful for future studies of chloroplast translation.

2 Current address: Department of Biology, University of Virginia, Charlottesville, VA 22901.

1 Supported in part by Grant 79-59-2361-1-1-327-1 from the United States Department of Agriculture Science and Education Administration Competitive Research Grants Office Program in Photosynthesis.






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