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Plant Molecular Biology

, Volume 38, Issue 5, pp 755–764 | Cite as

Isolation and characterization of two different cDNAs of Δ1-pyrroline-5-carboxylate synthase in alfalfa, transcriptionally induced upon salt stress

  • Idit Ginzberg
  • Hanan Stein
  • Yoram Kapulnik
  • Laszlo Szabados
  • Nicolai Strizhov
  • Jeff Schell
  • Csaba Koncz
  • Aviah Zilberstein
Article

Abstract

Two different cDNA clones, MsP5CS-1 and MsP5CS-2, encoding Δ1-pyrroline-5-carboxylate synthase (P5CS), the first enzyme of the proline biosynthetic pathway, were isolated from a λZap-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.

Medicago sativa proline pyrroline-5-carboxylate synthase salt stress 

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Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • Idit Ginzberg
    • 1
  • Hanan Stein
    • 1
  • Yoram Kapulnik
    • 2
  • Laszlo Szabados
    • 3
  • Nicolai Strizhov
    • 4
  • Jeff Schell
    • 4
  • Csaba Koncz
    • 3
  • Aviah Zilberstein
    • 1
  1. 1.Department of BotanyTel-Aviv UniversityTel-AvivIsrael
  2. 2.Institute of Field and Garden Crops, The Volcani CenterBet DaganIsrael
  3. 3.Institute of Plant Biology, Biological Research CenterHungarian Academy of SciencesSzegedHungary
  4. 4.Max-Planck InstituteKölnGermany

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