In the yeast Saccharomyces cerevisiae, the paralogous genes ALT1 and ALT2 have been proposed to encode alanine aminotransferase isozymes. Although in other microorganisms this enzyme constitutes the main pathway for alanine biosynthesis, its role in S. cerevisiae had remained unclear. Results presented in this paper show that under respiratory conditions, Alt1p constitutes the sole pathway for alanine biosynthesis and catabolism, constituting the first example of an alanine aminotransferase that simultaneously carries out both functions. Conversely, under fermentative conditions, it plays a catabolic role and alanine is mainly synthesized through an alternative pathway. It can thus be concluded that ALT1 has functions in alanine biosynthesis and utilization or only alanine utilization under respiratory and fermentative conditions, respectively. ALT2 expression was repressed under all tested conditions, suggesting that Alt2p biosynthesis is strictly controlled and only allowed to express under peculiar physiological conditions.
Little is known about the interactive effects of exogenous nitric oxide (NO) and abscisic acid (ABA) on nitrogen (N) metabolism and related changes at molecular and biochemical levels under drought stress. The present study highlights the independent and combined effect of NO and ABA (grouped as “nitrate agonists”) on expression profiles of representative key genes known to be involved in N-uptake and assimilation, together with proline metabolism, N–NO metabolism enzyme’s activity and nutrient content in polyethylene glycol (PEG) treated roots of Indian mustard (B. juncea cv. Varuna). Here we report that PEG mediated drought stress negatively inhibited growth performance, as manifested by reduced biomass (fresh and dry weight) production. Total N content and other nitrogenous compounds (NO3−, NO2−) were decreased; however, NH4+, NH4+/ NO3− ratio and total free amino acids content were increased. These results were positively correlated with the PEG induced changes in expression of genes and enzymes involved in N-uptake and assimilation. Also, PEG supply lowered the content of macro- and micro-nutrients but proline level and the activity of ∆1-pyrroline-5-carboxylate synthetase increased indicating increased oxidative stress. However, all these responses were reversed upon the exogenous application of nitrate agonists (PEG + NO, PEG + NO + ABA, and PEG + ABA) where NO containing nitrate agonist treatment i.e. PEG + NO was significantly more effective than PEG + ABA in alleviating drought stress. Further, increases in activities of L-arginine dependent NOS-like enzyme and S-nitrosoglutathione reductase were observed under nitrate agonist treatments. This indicates that the balanced endogenous change in NO and ABA levels together during synthesis and degradation of NO mitigated the oxidative stress in Indian mustard seedlings. Overall, our results reveal that NO independently or together with ABA may contribute to improved crop growth and productivity under drought stress.
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