Two maize genotypes differently responsive to nitrogen availability were characterized for their efficiency in nitrate accumulation via both the LATS (Low-Affinity Transport System) and HATS (High-Affinity Transport System) nitrate uptake systems. In addition, a full-length cDNA encoding a putative high-affinity nitrate transporter (ZmNrt2.1) was isolated and its expression evaluated in both the roots and leaves of the two maize genotypes, together with the expression of a maize H(+)-ATPase isoform (Mha1). The data showed the importance of the iHATS (Inducible High-Affinity System) system efficiency as a physiological marker of adaptation to low input and suggested that the transcript accumulation of ZmNrt2.1 might be a key step for the regulation of iHATS. However, ZmNrt2.1 transcription cannot account for the differences found between the two hybrids in terms of the activity of their respective iHATS and, as a consequence, of their adaptation to low input. Therefore, the involvement of some other transporter(s) or of some post-transcriptional/post-translational mechanism of regulation affecting the efficiency of iHATS may be hypothesized. In addition, the data suggest that the transcription of the Mha1 gene may also be involved in the global efficiency of the iHATS system.

Summary• The molecular properties and subcellular location of bound gamma-glutamyl transferase (GGT) were studied, and an experimental setup devised to assess its functions in barley roots.• Enzyme histochemistry was used to detect GGT activity at tissue level; immunocytochemistry to localize the protein at subcellular level; and modelling studies to investigate its surface charge properties. GGT activity in vivo was measured for the first time. Functions were explored by applying chemical treatments with inhibitors and the thiol-oxidizing drug diamide, performing time-course chromatographic and spectrophotometric analyses on low-molecular-weight thiols.• Gamma-glutamyl transferase activity was found to be high in the root apical region and the protein was anchored to root cell wall components, probably by basic amino acid residues. The results show that GGT is essential to the recovery of apoplastic glutathione provided exogenously or extruded by oxidative treatment.• It is demonstrated that GGT activity helps to salvage extracellular glutathione and may contribute to redox control of the extracellular environment, thus providing evidence of a functional role for gamma-glutamyl cycle in roots.

γ-Glutamyl transferases (GGT; EC 2.3.2.2) are glutathione-degrading enzymes that are represented in Arabidopsis thaliana by a small gene family of four members. Two isoforms, GGT1 and GGT2, are apoplastic, sharing broad similarities in their amino acid sequences, but they are differently expressed in the tissues: GGT1 is expressed in roots, leaves, and siliques, while GGT2 was thought to be expressed only in siliques. It is demonstrated here that GGT2 is also expressed in wild-type roots, albeit in very small amounts. GGT2 expression is enhanced in ggt1 knockout mutants, suggesting a compensatory effect to restore GGT activity in the root apoplast. Supplementation with 100 μM glutathione (GSH) resulted in the up-regulation of GGT2 gene expression in wild-type and ggt1 knockout roots, and of GGT1 gene expression in wild-type roots. Glutathione recovery was hampered by the GGT inhibitor serine/borate, suggesting a major role for apoplastic GGTs in this process. These findings can explain the ability of ggt1 knockout mutants to retrieve exogenously added glutathione from the growth medium.

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