The development of the activity of the chloroplastic and cytosolic isoenzymes of phosphoglycerate kinase during barley leaf ontogenesis

The catalytic activities of the chloroplastic and cytosolic isoenzymes of phosphoglycerate kinase (PGK; EC 2.7.2.3) have been followed during the development of the first leaf of barley (Hordeum vulgare L.) grown for 7 d in darkness followed by transfer to continuous illumination. The investigation has included both the study of a standard leaf section, measured from the leaf tip, over the whole life of the leaf and the study of serial sections of leaf, measured from the leaf base, at a standard sampling time. The results of both approaches were fully compatible. As the catalytic activity of each isoenzyme in the standard assay is directly proportional to the amount of isoenzyme protein present, the catalytic activities may be interpreted wholly in terms of enzyme synthesis and degradation. Both isoenzymes are synthesized in darkness and in etiolated barley are present at a ratio of about 2674 for the cytosolic to chloroplastic isoenzymes. Illumination results in a fivefold or greater increase in chloroplast PGK over a number of days with little change of the cytosolic isoenzyme, resulting in an eventual ratio of cytosolic to chloroplastic isoenzymes approaching the green-leaf value of about 991. Prior to any detectable onset of senescence a 15-fold increase in cytosolic isoenzyme commenced while the amount of chloroplast PGK remained constant. It is suggested that the increased cytosolic PGK may be involved in the export of carbohydrate reserves (starch) prior to leaf senescence. Both isoenzymes subsequently decline in parallel to total protein and chlorophyll in the course of senescence.Abbreviations DHAP reductase dihydroxyacetone-phosphate reductase - GS glutamine synthetase - LHCP light-harvesting chlorophyll-a/b-binding protein - PGK phosphoglycerate kinase - Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenaseThis work was supported by the Science and Engineering Research Council (grant no. GR/E54504).