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 lesbian hookup bars Savannah were tested for the GABA treatment, 100 µ m and 1 m m (Fig. 5). A positive and significant effect on 15 NO3 – 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 concentrations out of supply proteins was basically quantified for every single therapy so you can find out if GABA joined the underlying and dictate possible relationship between endogenous amino acid accounts and you may nitrate increase or Nrt2 mRNA top (Dining table step one). Immediately following a dozen h regarding exogenous supply, endogenous concentration of for every single amino acidic applied improved about resources. Particularly, step 1 meters meters GABA therapy enhanced root GABA because of the dos.24-fold weighed against the new handle herbs (Desk step one). However, a hundred µ meters GABA and you will step one meters meters Glu treatments were not successful to boost concentrations amounts of every four proteins used (Table step one). No correlation try found anywhere between transform regarding influx otherwise BnNrt2 transcript account and you will sources amino acid concentrations or ratio among them.
- Thinking found certainly are the means of around three separate replicates ± SD of your suggest.
Relationship ranging from glutamine within the phloem latex and you can nitrate consumption during Letter-deprivation as well as over the development period
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.
