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 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 density out-of sources proteins was in fact quantified per therapy so you’re able to verify that GABA inserted the underlying and to determine you are able to relationship ranging from endogenous amino acidic levels and you may nitrate increase otherwise Nrt2 mRNA height (Desk 1). After twelve h regarding exogenous have, endogenous concentration of for each amino acidic applied improved from the options. Such, 1 yards meters GABA medication improved means GABA by dos.24-bend compared to the fresh manage vegetation (Dining table step 1). not, one hundred µ yards GABA and step 1 m meters Glu solutions failed to boost density levels of most of the five amino acids applied (Table step one). Zero correlation is actually discovered anywhere between transform out-of increase or BnNrt2 transcript membership and you can root amino acid concentrations otherwise ratio between them.
- Thinking shown could be the manner of three separate replicates ± SD of one’s indicate.
Matchmaking between glutamine in phloem exudate and you may nitrate uptake throughout N-starvation as well as the organization years
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 https://datingranking.net/local-hookup/waco/ 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.
