Factors Involved in in Vitro Stabilization of Nitrate Reductase from Cotton (Gossypium hirsutum L.) Cotyledons

Experiments were conducted to determine if pretreatment of cotton (Gossypium hirsutum L.) plants resulted in differential in vitro stabilities of nitrate reductase (NR) activity. Although NR activity declines markedly during the second half of the daily light period, in vitro NR stability is not modified by time of harvest. Phenylmethylsulfonylfluoride, iodoacetamide, and N-ethylmaleimide do not influence in vitro NR stability, suggesting that serine or sulfhydryl proteases are not responsible for in vitro lability of NR from cotton cotyledons.Imposition of water stress or artificial extension of the dark period lead to significant reductions in NR activity, but do not change in vitro NR stability.Dilution of a crude extract leads to increasing lability of NR; hence the marked instability of NR cannot be attributed to an inactivator which follows simple enzyme kinetics. Since in vitro NR activity is much more stable in presence of both NADH and NO(3) (-), substrate availability must be considered as a possible factor influencing in vivo NR stability.