Distribution of Nitrate-assimilating Enzymes between Mesophyll Protoplasts and Bundle Sheath Cells in Leaves of Three Groups of C4 Plants

Abstract

Intercellular distribution of enzymes involved in amino N synthesis was studied in leaves of species representing 3 C4 groups, i.e., Sorghum bicolor, Zea mays, Digitaria sanguinalis (NADP Malic enzyme type); Panicum miliaceum (NAD malic enzyme type); and P. maximum (phosphoenolpyruvate carboxykinase type). Nitrate reductase, nitrite reductase, glutamine synthetase and glutamate synthase were predominantly localized in mesophyll cells of all the species, except in P. maximum where nitrite reductase had similar activity on a chlorophyll basis, in both mesophyll and bundle sheath cells. NADH-glutamate dehydrogenase was concentrated in the bundle sheath cells, while NADPH-glutamate dehydrogenase was localized in both mesophyll and bundle sheath cells. The activities of nitrate-assimilating enzymes, except for nitrate reductase, were high enough to account for the proposed in vivo rates of nitrate assimilation. Based on the differential centrifugation of cell homogenates of P. miliaceum, mesophyll chloroplasts appear to be the major site of nitrate assimilation since nitrite reductase, glutamine synthetase, glutamate synthase, and NADPH-glutamate dehydrogenase were primarily localized in the chloroplast fraction. Both the glutamine synthetase-glutamate synthase and glutamate dehydrogenase pathways were considered as alternative routes of amino N synthesis.