Cytosolic glutamine synthetase in soybean is encoded by a multigene family, and the members are regulated in an organ-specific and developmental manner

  • Morey K
  • Ortega J
  • Sengupta-Gopalan C
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Abstract

Gln synthetase (GS) is the key enzyme in N metabolism and it catalyzes the synthesis of Gln from glutamic acid, ATP, and NH4+. There are two major isoforms of GS in plants, a cytosolic form (GS1) and a chloroplastic form (GS2). In leaves, GS2 functions to assimilate ammonia produced by nitrate reduction and photorespiration, and GS1 is the major isoform assimilating NH3 produced by all other metabolic processes, including symbiotic N2 fixation in the nodules. GS1 is encoded by a small multigene family in soybean (Glycine max), and cDNA clones for the different members have been isolated. Based on sequence divergence in the 3'-untranslated region, three distinct classes of GS1 genes have been identified (alpha, beta, and gamma). Genomic Southern analysis and analysis of hybrid-select translation products suggest that each class has two distinct members. The alpha forms are the major isoforms in the cotyledons and young roots. The beta forms, although constitutive in their expression pattern, are ammonia inducible and show high expression in N2-fixing nodules. The gamma1 gene appears to be more nodule specific, whereas the gamma2 gene member, although nodule enhanced, is also expressed in the cotyledons and flowers. The two members of the alpha and beta class of GS1 genes show subtle differences in the expression pattern. Analysis of the promoter regions of the gamma1 and gamma2 genes show sequence conservation around the TATA box but complete divergence in the rest of the promoter region. We postulate that each member of the three GS1 gene classes may be derived from the two ancestral genomes from which the allotetraploid soybean was derived.

Author-supplied keywords

  • *Multigene Family
  • Adenosine Triphosphate/metabolism
  • Ammonia/metabolism
  • Blotting, Southern
  • Cotyledon/enzymology/genetics
  • Cytosol/enzymology
  • DNA, Complementary/chemistry/genetics/isolation &
  • Gene Expression Regulation, Enzymologic
  • Gene Expression Regulation, Plant
  • Glutamate-Ammonia Ligase/*genetics/metabolism
  • Glutamic Acid/metabolism
  • Glutamine/biosynthesis
  • Isoenzymes/genetics/metabolism
  • Molecular Sequence Data
  • Nitrates/metabolism
  • Nitrogen/metabolism
  • Phylogeny
  • Plant Components/enzymology/*genetics
  • Plant Roots/enzymology/genetics
  • Promoter Regions (Genetics)/genetics
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Seeds/enzymology/genetics
  • Sequence Analysis, DNA
  • Soybeans/enzymology/*genetics
  • TATA Box/genetics

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Authors

  • K J Morey

  • J L Ortega

  • C Sengupta-Gopalan

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