Differential expression of the two cytosolic glutamine synthetase genes in various organs of Medicago truncatula

In order to clarify the physiological roles of the cytosolic forms of glutamine synthetase (GS) in Medicago truncatula, we have performed a detailed analysis of the expression of the two functional cytosolic GS genes, MtGSa and MtGSb in several organs of the plant. Transcriptional fusions were made between the 2.6 or 3.1 kbp 5' upstream regions of MtGSa or MtGSb, respectively, and the reporter gene gusA encoding beta -glucuronidase and introduced into the homologous transgenic system. MtGSa and MtGSb were found to be differentially expressed in most of the organs, both temporally and spatially. The presence of GS proteins at the sites where the promoters were active was confirmed by immunocytochemistry, providing the means to correlate gene expression with the protein products. These studies have shown that the putative MtGSa and MtGSb promoter fragments were sufficient to drive GUS expression in all the tissues and cell types where cytosolic GS proteins were located. This result indicates that the cis acting regulatory elements responsible for conferring the contrasting expression patterns are located within the region upstream of the coding sequences. MtGSa was preferentially expressed in the vascular tissues of almost all the organs examined, whereas MtGSb was preferentially expressed in the root cortex and in leaf pulvini. The location and high abundance of GS in the vascular tissues of almost all the organs analysed suggest that the enzyme encoded by MtGSa plays an important role in the production of nitrogen transport compounds. The enzyme synthesised by MtGSb appears to have more ubiquitous functions for ammonium assimilation and detoxification in a variety of organs. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved.