Differential effects of nitrate and ammonium supply on nodule initiation, development, and distribution on roots of pea (Pisum sativum)

Abstract

Low, static concentrations of ammonium have less negative effects on nodulation of pea ( Pisum sativum L.) than nitrate and in some cases may actually stimulate nodulation. Two experiments were carried out to assess the effects of supplying both forms of mineral N, separately and in combination, on nodule initiation, nodule development, nodule distribution between primary and lateral (secondary) roots, tertiary root development, and N₂ fixation in pea. Pea plants were grown for up to 24 d after inoculation in hydroponic culture with no mineral N (zero N), NO₃^– (0.5 mmol·L^–1), NH₄⁺ (0.5 mmol·L^–1), or NO₃^– (0.25 mmol·L^–1) plus NH₄⁺ (0.25 mmol·L^–1). Concentrations of nitrate and ammonium were monitored on a daily basis and held relatively constant by continuous, automatic additions of stock solutions. Pea plants accumulated the most total dry mass (DM) and total N when supplied with the combination of nitrate plus ammonium but had the lowest nodule DM and percentage of nitrogen derived from the atmosphere. Whole-plant nodulation (nodules per plant) and DM-specific nodulation (nodules·g^–1 root DM) were 2.3- and 2.4-fold greater, respectively, in pea plants receiving NH₄⁺ at 0.5 mmol·L^–1 than in those supplied with NO₃^– at 0.5 mmol·L^–1. The nodulation responses of plants receiving NO₃^– at 0.25 mmol·L^–1 plus NH₄⁺ at 0.25 mmol·L^–1 were more similar to those of plants receiving only nitrate than only ammonium, indicating that when both forms of mineral N are available to plants, nitrate has a predominant effect on the nodulation response. Assessment of the stage of development of nodule primordia and nodules during the time course of the experiments indicated that nitrate not only decreased the degree of nodule initiation but also the rate at which those nodules developed. Microscopic observations indicated that the more negative effects of the nitrate treatment on DM-specific nodulation as compared with the ammonium treatment were consistent on both the primary and lateral roots. Quantification of nodulation and tertiary root development on lateral roots indicated that the stimulating effects of ammonium were specific to nodulation; the effects on tertiary root development were different. The study demonstrates for the first time that when both forms of mineral N are available at equal concentrations, the nodulation response in pea is influenced more by nitrate than by ammonium and that the effects of nitrate and ammonium on tertiary root initiation and development are unlike those on nodulation.