Differential influence of herbicide treatments on activity and kinetic parameters of C4 photosynthetic enzymes from Zea mays

The recommended field dose of rimsulfuron, imazethapyr, alachlor, atrazine or fluometuron differentially reduced shoot fresh and dry weight of 10-day-old maize seedlings as well as leaf protein content during the following 12 days. These reductions seemed consistent during the whole period with fluometuron, atrazine and alachlor but appeared to be nullified by the 5th day of treatment with rimsulfuron and imazethapyr. On the other hand, all herbicides mostly provoked significant inhibitions in specific activities of phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31), malate dehydrogenase (MDH, EC 1.1.1.82), pyruvate phosphate dikinase (PPDK, EC 2.7.9.1) and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco, EC 4.1.1.39) in leaves during the first 2 days. Thereafter, the inhibition was recovered in samples treated with rimsulfuron and imazethapyr, leveled off with alachlor but consistently augmented with atrazine and fluometuron. The kinetic characterization showed that rimsulfuron or imazethapyr unchanged V-max of all enzymes in vitro, however, V-max of PEPC, PPDK and Rubisco were decreased in vivo. Nevertheless, atrazine or fluometuron substantially reduced V-max of all enzymes while alachlor showed a reduction in this value of PEPC, MDH and Rubisco. Thus atrazine, fluometuron and, to a lower extent, alachlor reduced concentrations of all enzymes as well as rimsulfuron and imazethapyr for only Rubisco. On the contrary, K-m values of all enzymes were progressively increased by all herbicides indicating that the different herbicides altered the structural integrity of all enzymes. These findings conclude that rimsulfuron or imazethapyr competitively inhibited MDH but revealed mixed inhibition to PEPC, PPDK and Rubisco. Atrazine or fluometuron revealed mixed inhibitions to all enzymes whereas alachlor seemed to be either a competitive inhibitor to PPDK or a mixed inhibitor to PEPC, MDH and Rubisco. (C) 2007 Elsevier Inc. All rights reserved.