Gene transcript accumulation and in situ mRNA hybridization of two putative glutamate dehydrogenase genes in etiolated Glycine max seedlings.
Glutamate dehydrogenase (EC 1.4.1.2) is a multimeric enzyme that catalyzes the reversible amination of α-ketoglutarate to form glutamate. We characterized cDNA clones of two Glycine max sequences, GmGDH1 and GmGDH2, that code for putative α- and β-subunits, respectively, of the NADH dependent enzyme. Temporal and spatial gene transcript accumulation studies using semiquantitative RT-PCR and in situ hybridization have shown an overlapping gene transcript accumulation pattern with differences in relative gene transcript accumulation in the organs examined. Detection of NADH-dependent glutamate dehydrogenase activity in situ using a histochemical method showed concordance with the spatial gene transcript accumulation patterns. Our findings suggest that although the two gene transcripts are co-localized in roots of etiolated soybean seedlings, the ratio of the two subunits of the active holoenzyme may vary among tissues.
Laboratory of General and Agricultural Microbiology, Agricultural University of Athens , Athens , Greece.
M
M
Dimou
G
G
Tsaniklidis
G
G
Aivalakis
P
P
Katinakis
D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D017343:Genes, Plant; D005969:Glutamate Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D006651:Histocytochemistry; D017403:In Situ Hybridization; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D018517:Plant Roots / Q000201:enzymology / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D018749:RNA, Plant / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D036226:Seedlings / Q000201:enzymology / Q000235:genetics; D013025:Glycine max / Q000201:enzymology* / Q000235:genetics*
D018076:DNA, Complementary; D018744:DNA, Plant; D007527:Isoenzymes; D010940:Plant Proteins; D012333:RNA, Messenger; D018749:RNA, Plant; D005969:Glutamate Dehydrogenase
Glycine max; NADH glutamate dehydrogenase; RT-PCR; in situ hybridization; root