In order to test the hypothesis that GABA could up-regulate nitrate uptake, the effect of exogenous supply of different amino acids was compared with control plants. Gln, Glu and Asn are known to act as inhibitors of nitrate uptake at the millimolar range in many species when supplied to the root nutrient solution ( Muller & Touraine 1992 ; Vid ). Therefore, roots were supplied with either 1 m m Gln, 1 m m Glu or 1 m m Asn, whereas two concentrations were tested for the GABA treatment, 100 µ m and 1 m m (Fig. 5). A positive and significant effect on 15 NOstep three – influx was observed for 100 µ m (P < 0.05) and no significant effect for 1 m m GABA (P > 0.05)-treated plants compared with the control plants, whereas 15 NO3 – influx was significantly reduced by the Gln, Glu and Asn root treatments (Fig. 5a). The strongest negative effect on nitrate uptake was due to Glu (38% inhibition) followed by Asn (37% inhibition) and Gln (33% inhibition). In parallel, Northern blot analyses performed with mRNA extracted from the same amino acid treated plants confirmed previous results for all of the amino acids tested. Thus, BnNrt2 mRNA abundance was enhanced by 1.6- and 1.3-fold in response to 100 µ m and 1 m m GABA treatments, respectively. The Gln, Glu how to hookup in Toledo and Asn treatments reduced BnNrt2 mRNA abundance by 0.5-, 0.9- and 0.8-fold, respectively (Figs 5b & c).
Effects of exogenous supply of GABA, Gln, Glu and Asn on nitrate influx (a) and BnNrt2 gene expression (b and c) on 12 h nitrate deprived plants. Plants were supplied with 1 m m nitrate plus either 100 µ m or 1 m m GABA, 1 m m Gln, 1 m m Glu or 1 m m Asn for 12 h before measurement of 15 NO3 – influx and BnNrt2 transcript abundance. Influx and BnNrt2 transcript expression values are the means of six and two pots of 25 plants, respectively. Vertical bars indicate ± SE of the mean for n = 6 when larger than the symbol.
Endogenous levels from means proteins had been quantified per cures in order to find out if GABA entered the underlying and also to dictate you’ll be able to relationship anywhere between endogenous amino acidic profile and you may nitrate increase otherwise Nrt2 mRNA peak (Dining table step 1). Immediately following a dozen h from exogenous also have, endogenous intensity of each amino acidic used increased from the sources. For example, 1 m m GABA cures enhanced sources GABA by the 2.24-bend compared with the new control flowers (Table step 1). Although not, 100 µ m GABA and you may step one meters yards Glu solutions failed to improve levels amounts of all four proteins applied (Dining table step 1). Zero correlation try found ranging from change of influx or BnNrt2 transcript membership and sources amino acid density or proportion between them.
- Thinking shown are definitely the manner of three independent replicates ± SD of suggest.
Relationship ranging from glutamine in phloem exudate and you can nitrate consumption during the N-deprivation as well as over the development cycle
It is well established that N-deprivation leads to a transient ‘de-repression’ of high-affinity nitrate influx and Nrt2 gene expression in plants with a low N status ( Lejay et al. 1999 ; Forde 2002 ). In plants with a high N status, ‘de-repression’ seems to be bypassed, leading to a direct decline of nitrate uptake interpreted as ‘de-induction’ ( Clarkson 1986 ; Siddiqi et al. 1989 ; Faure-Rabasse et al. 2002 ). Our results for 15 NO3 – influx and BnNrt2 gene expression (Fig. 1a & c) are consistent with these previous studies.