ABSTRACT
A de-repression mechanism based on the disappearance of 'signals' down-regulating N transporter activity has been proposed in the literature to explain the transient increase of NO(3)(-) uptake by the roots following N deprivation in higher plants. This hypothesis was investigated at the physiological and molecular levels by measuring NO(3)(-) influx into roots of Brassica napus L. grown under low or high external concentrations of KNO(3) following N deprivation. Parallel measurements were made of endogenous NO(3)(-), amino acid concentrations and abundance of mRNA for BnNRT1 and BnNRT2, genes encoding nitrate-inducible transport proteins. The effect of NO(3)(-) pulsing on NO(3)(-) transport components in N-deprived plants was also investigated by measuring influx of high- and low-affinity transport system (HATS and LATS) and assaying mRNA levels. Influx of NO(3)(-) via HATS and LATS, and transcript levels of BnNRT2 and BnNRT1 decreased with the duration of N deprivation. The results suggested that the absence of de-repression of NO(3)(-) influx and BnNRT2 gene expression following N starvation was related to a high amino acid status. Pulsing with NO(3)(-) induced a large increase in BnNRT2 mRNA level, but a comparatively small increase in NO(3)(-) influx via HATS. The level of BnNRT1 mRNA also increased, but there was no effect on LATS uptake activity. The absence of a strict correlation between the NO(3)(-) transport activity and the mRNA BnNRT1 and BnNRT2 levels is discussed in terms of possible post-transcriptional regulation by the amino acids.
Subject(s)
Anion Transport Proteins/metabolism , Arabidopsis Proteins/metabolism , Brassica napus/genetics , Nitrates/pharmacology , Nitrogen/deficiency , Plant Proteins , Amino Acids/metabolism , Anion Transport Proteins/drug effects , Anion Transport Proteins/genetics , Arabidopsis Proteins/drug effects , Arabidopsis Proteins/genetics , Biological Transport, Active/drug effects , Blotting, Northern , Brassica napus/drug effects , Brassica napus/metabolism , Cloning, Molecular , Gene Expression Regulation, Plant/drug effects , Glutamic Acid/metabolism , Glutamine/metabolism , Kinetics , Nitrate Transporters , Nitrates/metabolism , Nitrogen/pharmacology , Nitrogen Isotopes , Plant Roots/drug effects , Plant Roots/metabolism , Plant Shoots/drug effects , Plant Shoots/metabolism , RNA, Messenger/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism , Signal Transduction/drug effectsABSTRACT
A simple three equation model is proposed for the feedback regulation of nitrate uptake and N2 fixation, based on the concentration of the organic N substrate pool within the plant and two parameters denoting the N substrate concentrations at which half-maximal inhibition occurs. This model simulated three contrasting phenotypes of white clover (Trifolium repens L.) inbred lines with (1) normal rates of nitrate uptake and N2 fixation (NNU); (2) low rates of nitrate uptake (LNU); and (3) very low rates of N2 fixation (VLF). The LNU phenotype was simulated by a decrease in the value of the inhibition parameter for nitrate uptake and the VLF phenotype was simulated by a decrease in the value of the N2 fixation inhibition parameter. The model was tested against nitrate uptake data obtained from white clover plants growing in flowing nutrient culture. There was an accurate prediction of the increase in nitrate uptake caused by N2 fixation activity of the NNU and LNU inbred lines being interrupted by a switch in gas phase from air to Ar : O2. The model was also tested against data for nitrate uptake, N2 fixation and %N from fixation for the three inbred clover lines grown in flowing nutrient culture at 0, 5 or 20 mmol m(-3) N(3-). Again there was accurate prediction of nitrate uptake, although simulated values for N2 fixation were more variable. The simple model has potential use as a sub-routine in larger models of legume growth under field conditions.