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 NO_{step step step 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 and Asn treatments reduced BnNrt2 mRNA abundance by 0.5-, 0.9- and 0.8-fold, respectively (Figs 5b & c).

Dialogue

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 density regarding sources amino acids had been quantified for each procedures to help you verify that GABA joined the underlying and also to dictate you are able to correlation between endogenous amino acidic hookup clubs Sheffield membership and you can nitrate influx or Nrt2 mRNA peak (Dining table 1). Once several h away from exogenous supply, endogenous concentration of for every single amino acidic applied enhanced regarding sources. Including, 1 meters yards GABA treatment improved means GABA by the dos.24-flex in contrast to the manage herbs (Table 1). Although not, one hundred µ m GABA and you will 1 yards m Glu treatments failed to boost levels degrees of all the five proteins applied (Desk step 1). Zero relationship is actually located anywhere between alter of influx or BnNrt2 transcript account and you can sources amino acidic concentrations otherwise ratio between them.

• Values shown is the technique of around three separate replicates ± SD of one’s imply.

Relationship anywhere between glutamine within the phloem exudate and you will nitrate use during N-starvation as well as over the organization duration

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.