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 https://datingranking.net/local-hookup/tucson/ 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).
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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 away from supply proteins was in fact quantified for every medication so you can find out if GABA entered the underlying and also to determine you are able to correlation anywhere between endogenous amino acid account and you may nitrate increase or Nrt2 mRNA top (Desk 1). After a dozen h away from exogenous also have, endogenous intensity of for each and every amino acidic applied increased in the root. Including, step 1 m meters GABA medication increased supply GABA by the dos.24-flex compared with the latest control vegetation (Dining table step 1). not, one hundred µ yards GABA and 1 yards yards Glu solutions were not successful to boost density degrees of all of the four amino acids used (Table 1). No correlation try located anywhere between change from increase or BnNrt2 transcript levels and options amino acid levels otherwise ratio among them.
- Opinions found are the a style of around three independent replicates ± SD of your own imply.
Relationships ranging from glutamine inside the phloem exudate and you will nitrate consumption through the N-starvation and over the growth 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.
