Abstract
BackgroundNitrogen uptake and the efficient absorption and metabolism of nitrogen are essential elements in attempts to breed improved cereal cultivars for grain or silage production. One of the enzymes related to nitrogen metabolism is glutamine-2-oxoglutarate amidotransferase (GOGAT). Together with glutamine synthetase (GS), GOGAT maintains the flow of nitrogen from NH4 + into glutamine and glutamate, which are then used for several aminotransferase reactions during amino acid synthesis.ResultsThe aim of the present work was to identify and analyse the structure of wheat NADH-GOGAT genomic sequences, and study the expression in two durum wheat cultivars characterized by low and high kernel protein content. The genomic sequences of the three homoeologous A, B and D NADH-GOGAT genes were obtained for hexaploid Triticum aestivum and the tetraploid A and B genes of Triticum turgidum ssp. durum. Analysis of the gene sequences indicates that all wheat NADH-GOGAT genes are composed of 22 exons and 21 introns. The three hexaploid wheat homoeologous genes have high conservation of sequence except intron 13 which shows differences in both length and sequence. A comparative analysis of sequences among di- and mono-cotyledonous plants shows both regions of high conservation and of divergence. qRT-PCR performed with the two durum wheat cvs Svevo and Ciccio (characterized by high and low protein content, respectively) indicates different expression levels of the two NADH-GOGAT-3A and NADH-GOGAT-3B genes.ConclusionThe three hexaploid wheat homoeologous NADH-GOGAT gene sequences are highly conserved – consistent with the key metabolic role of this gene. However, the dicot and monocot amino acid sequences show distinctive patterns, particularly in the transit peptide, the exon 16–17 junction, and the C-terminus. The lack of conservation in the transit peptide may indicate subcellular differences between the two plant divisions - while the sequence conservation within enzyme functional domains remains high. Higher expression levels of NADH-GOGAT are associated with higher grain protein content in two durum wheats.
Highlights
Nitrogen uptake is an essential element in crop improvement, either directly for grain protein content or indirectly for photosynthetic production
By comparing this contig with the reported chromosome 3B genomic sequence containing a NADHGOGAT gene (FN564429) and the sequence obtained by assembling D-genome 454-sequences, we were able to separate the 487 Chinese Spring 454-reads into three sequences assignable to each of the three genomes: 178 reads to the A-genome, 162 to the B-genome, and 147 to the D-genome
The three hexaploid wheat homoeologous NADH-glutamine-2-oxoglutarate amidotransferase (GOGAT) gene sequences are given in File S1
Summary
Nitrogen uptake is an essential element in crop improvement, either directly for grain protein content or indirectly for photosynthetic production. Despite that GY and GPC are genetically negatively correlated, it has been shown in various cereals (including wheat) that this correlation can be broken down by adequate nitrogen supply in late plant development [1,2]. This requires that growers must optimize the use of nitrogen fertilizers to avoid pollution, while maintaining reasonable profit margins. Together with glutamine synthetase (GS), GOGAT maintains the flow of nitrogen from NH4+ into glutamine and glutamate, which are used for several aminotransferase reactions during amino acid synthesis
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