Isolation of the ornithine-delta-aminotransferase cDNA and effect of salt stress on its expression in Arabidopsis thaliana.
To evaluate the relative importance of ornithine (Orn) as a precursor in proline (Pro) synthesis, we isolated and sequenced a cDNA encoding the Orn-delta-aminotransferase (delta-OAT) from Arabidopsis thaliana. The deduced amino acid sequence showed high homology with bacterial, yeast, mammalian, and plant sequences, and the N-terminal residues exhibited several common features with a mitochondrial transit peptide. Our results show that under both salt stress and normal conditions, delta-OAT activity and mRNA in young plantlets are slightly higher than in older plants. This appears to be related to the necessity to dispose of an easy recycling product, glutamate. Analysis of the expression of the gene revealed a close association with salt stress and Pro production. In young plantlets, free Pro content, Delta1-pyrroline-5-carboxylate synthase mRNA, delta-OAT activity, and delta-OAT mRNA were all increased by salt-stress treatment. These results suggest that for A. thaliana, the Orn pathway, together with the glutamate pathway, plays an important role in Pro accumulation during osmotic stress. Conversely, in 4-week-old A. thaliana plants, although free Pro level also increased under salt-stress conditions, the delta-OAT activity appeared to be unchanged and delta-OAT mRNA was not detectable. Delta1-pyrroline-5-carboxylate synthase mRNA was still induced at a similar level. Therefore, for the adult plants the free Pro increase seemed to be due to the activity of the enzymes of the glutamate pathway.
Roosens
N H
NH
Laboratorium voor Plantengenetica, Vrije Universiteit Brussel, Paardenstraat 65, Sint-Genesius-Rode, B-1640 Belgium. nroosens@vub.ac.be
Thu
T T
TT
Iskandar
H M
HM
Jacobs
M
M
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis / Q000302:isolation & purification*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D008969:Molecular Sequence Data; D009953:Ornithine-Oxo-Acid Transaminase / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D009994:Osmolar Concentration; D011392:Proline / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology
D018076:DNA, Complementary; D012965:Sodium Chloride; D011392:Proline; D009953:Ornithine-Oxo-Acid Transaminase