Reevaluation of the “in vitro” and “in vivo” methods of nitrate reductase determination in pea roots

Abstract

Nitrate consumption, nitrite production and nitrite loss were followed in the“in vitro” nitrate reductase (NR) assay mixtures with pea root extracts, and nitrite production in the “in vitro” NR assay was compared with anaerobic nitrite production in the “in vivo” assay with pea root segments. NR activity could not be reliably determined on the basis of nitrate consumption using three methods of NO3-determination (salicylate, cadmium, enzymatic). NR activity based on nitrite determination was slightly underestimated due to nitrite loss (less than 10% with pea root extracts) in the assay mixture. Nitrite production in the “in vivo” NR assay under optimum conditions closely resembled nitrite production in the “in vitro” NR assay and thus reflected NR level in the tissue.