Isolation and characterization of two different cDNAs of delta1-pyrroline-5-carboxylate synthase in alfalfa, transcriptionally induced upon salt stress

Two different cDNA clones, MsP5CS-1 and MsP5CS-2, encoding delta1 -pyrroline-5-carboxylate synthase (P5CS). the first enzyme of the proline biosynthetic pathway, were isolated from a lambdaZap-cDNA library constructed from salt stressed Medicago sativa roots. MsP5CS-1 (2.6 kb) has an open reading fr...

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Bibliographic Details
Published in:Plant molecular biology 1998-11, Vol.38 (5), p.755-764
Main Authors: Ginzberg, I, Stein, H, Kapulnik, Y, Szabados, L, Strizhov, N, Schell, J, Koncz, C, Zilberstein, A
Format: Article
Language:English
Subjects:
1-Pyrroline-5-Carboxylate Dehydrogenase
Amino Acid Sequence
Base Sequence
DNA, Complementary - chemistry
DNA, Complementary - genetics
DNA, Complementary - isolation & purification
Gene Expression Regulation, Enzymologic
Gene Expression Regulation, Plant
Medicago sativa - chemistry
Medicago sativa - enzymology
Medicago sativa - genetics
Molecular Sequence Data
Oxidoreductases Acting on CH-NH Group Donors - genetics
Plant Roots - chemistry
Plant Roots - drug effects
Plant Roots - metabolism
Proline - drug effects
Proline - metabolism
Sequence Analysis, DNA
Sequence Homology, Amino Acid
Sequence Homology, Nucleic Acid
Sodium Chloride - pharmacology
Space life sciences
Transcription, Genetic - drug effects
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Summary:Two different cDNA clones, MsP5CS-1 and MsP5CS-2, encoding delta1 -pyrroline-5-carboxylate synthase (P5CS). the first enzyme of the proline biosynthetic pathway, were isolated from a lambdaZap-cDNA library constructed from salt stressed Medicago sativa roots. MsP5CS-1 (2.6 kb) has an open reading frame of 717 amino acids, as well as a non-spliced intron at a position corresponding to the evolutionary fusion point of the bacterial proA and proB genes. MsP5CS-2 (1.25 kb) is a partial clone. The clones share 65% identity in nucleotide sequences, 74% homology in deduced amino acid sequences, and both show a high similarity to Vigna aconitifolia and Arabidopsis thaliana P5CS cDNA clones. Southern blot analysis confirmed the presence of two different P5CS genes. The effect of salinity on the transcription of MsP5CS-1 and MsP5CS-2 in roots was studied, using northern blot analysis and a RT-PCR approach. A rapid increase in the steady-state transcript level of both genes in roots was observed by RT-PCR upon exposure of hydroponically grown 6-day old seedlings to 90 mM NaCl, suggesting that both are salt-inducible genes, yet a higher response was observed for MsP5CS-2.
ISSN:0167-4412