Overexpression of Δ1-pyrroline-5-carboxylate synthetase increases proline production and confers salt tolerance in transgenic potato plants

Abstract Proline (Pro) is known as compatible osmolyte accumulated by plant cells in response to salt and drought stresses. It is supposed to be an osmoprotectant involved in the protection of cellular structures under osmotic stress. Therefore, in an attempt to increase salt tolerance in potato, a pyrroline-5-carboxylate synthetase (P5CS) cDNA from Arabidopsis thaliana was transferred to potato plants via Agrobacterium-mediated transformation. This enzyme is responsible for conversion of glutamate to Δ1-pyrroline-5-carboxylate that is reduced to Pro. The resulting transgenic potato plants showed an important increase in Pro production levels compared to non-transgenics. This Pro accumulation was particularly enhanced in the presence of salt up to 100 mM NaCl. The transgenic potato plants also showed an improved tolerance to salinity through a much less altered tuber yield and weight compared to the non-transgenic ones.

[1]  F. Neidhardt,et al.  Escherichia Coli and Salmonella: Typhimurium Cellular and Molecular Biology , 1987 .

[2]  P. Saradhi,et al.  Proline accumulation under heavy metal stress , 1991 .

[3]  W. Sawahel,et al.  Generation of transgenic wheat plants producing high levels of the osmoprotectant proline , 2002, Biotechnology Letters.

[4]  A. Hanson,et al.  Metabolic Responses of Mesophytes to Plant Water Deficits , 1982 .

[5]  R. Munns,et al.  Mechanisms of salt tolerance in nonhalophytes. , 1980 .

[6]  D. Verma,et al.  Subcellular location of delta-pyrroline-5-carboxylate reductase in root/nodule and leaf of soybean. , 1992, Plant physiology.

[7]  A. Moftah,et al.  The effect of sodium chloride on solute potential and proline accumulation in soybean leaves. , 1987, Plant physiology.

[8]  K. Shinozaki,et al.  Correlation between the induction of a gene for delta 1-pyrroline-5-carboxylate synthetase and the accumulation of proline in Arabidopsis thaliana under osmotic stress. , 1995, The Plant journal : for cell and molecular biology.

[9]  C. Hagedorn,et al.  Proline metabolism in N2-fixing root nodules: energy transfer and regulation of purine synthesis. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[10]  R. Munns,et al.  SOLUTE ACCUMULATION IN THE APEX AND LEAVES OF WHEAT DURING WATER-STRESS , 1979 .

[11]  G. Fabro,et al.  Proline accumulation and AtP5CS2 gene activation are induced by plant-pathogen incompatible interactions in Arabidopsis. , 2004, Molecular plant-microbe interactions : MPMI.

[12]  L N Csonka,et al.  Physiological and genetic responses of bacteria to osmotic stress. , 1989, Microbiological reviews.

[13]  D. Verma,et al.  Characterization of Δ1-pyrroline-5-carboxylate synthetase gene promoter in transgenic Arabidopsis thaliana subjected to water stress , 1997 .

[14]  G. Ducreux,et al.  Production and characterization of intergeneric somatic hybrids through protoplast electrofusion between potato (Solanum tuberosum) and Lycopersicon pennellii , 1994, Plant Cell, Tissue and Organ Culture.

[15]  James B. Hicks,et al.  A plant DNA minipreparation: Version II , 1983, Plant Molecular Biology Reporter.

[16]  Marc Van Montagu,et al.  Efficient octopine Ti plasmid-derived vectors for Agrobacterium- mediated gene transfer to plants , 1985, Nucleic Acids Res..

[17]  P. Verslues,et al.  Proline accumulation in maize (Zea mays L.) primary roots at low water potentials. II. Metabolic source of increased proline deposition in the elongation zone , 1999, Plant physiology.

[18]  S. -. Park,et al.  Improvement of salt tolerance in transgenic potato plants by glyceraldehyde-3 phosphate dehydrogenase gene transfer. , 2001, Molecules and cells.

[19]  D. Verma,et al.  A bifunctional enzyme (delta 1-pyrroline-5-carboxylate synthetase) catalyzes the first two steps in proline biosynthesis in plants. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[20]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[21]  C. Hwang,et al.  Research Articles : Molecular Biology/Gene Transformation ; Salt Tolerance Enhanced by Transformation of a P5CS Gene in Carrot , 2003 .

[22]  E. Ábrahám,et al.  Differential expression of two P5CS genes controlling proline accumulation during salt-stress requires ABA and is regulated by ABA1, ABI1 and AXR2 in Arabidopsis. , 1997, The Plant journal : for cell and molecular biology.

[23]  J. Christian The Influence of Nutrition on the Water Relations of Salmonella oranienburg. , 1955 .

[24]  A. Huang,et al.  Proline Oxidase and Water Stress-induced Proline Accumulation in Spinach Leaves. , 1979, Plant physiology.

[25]  K. Shinozaki,et al.  Stress-responsive and developmental regulation of Delta(1)-pyrroline-5-carboxylate synthetase 1 (P5CS1) gene expression in Arabidopsis thaliana. , 1999, Biochemical and biophysical research communications.

[26]  H. Bohnert,et al.  Increased Resistance to Oxidative Stress in Transgenic Plants by Targeting Mannitol Biosynthesis to Chloroplasts , 1997, Plant physiology.

[27]  C. Stewart Inhibition of proline oxidation by water stress. , 1977, Plant physiology.

[28]  R. Bressan,et al.  Metabolic changes associated with adaptation of plant cells to water stress. , 1986, Plant physiology.

[29]  N. Smirnoff,et al.  Hydroxyl radical scavenging activity of compatible solutes , 1989 .

[30]  M. Van Montagu,et al.  Isolation, characterization, and chromosomal location of a gene encoding the Δ 1‐pyrroline‐5‐carboxylate synthetase in Arabidopsis thaliana , 1995, FEBS letters.

[31]  J. Venekamp,et al.  The sources of free proline and asparagine in field bean plants, Vicia faba L., during and after a short period of water withholding , 1988 .

[32]  L. Brown,et al.  Sorbitol and proline as intracellular osmotic solutes in the green alga Stichococcus bacillaris , 1978 .

[33]  A. Hanson,et al.  Prokaryotic osmoregulation: genetics and physiology. , 1991, Annual review of microbiology.

[34]  D. Verma,et al.  Cloning of ornithine delta-aminotransferase cDNA from Vigna aconitifolia by trans-complementation in Escherichia coli and regulation of proline biosynthesis. , 1993, The Journal of biological chemistry.

[35]  D. Verma,et al.  PROLINE BIOSYNTHESIS AND OSMOREGULATION IN PLANTS , 1993 .

[36]  Z. Peng,et al.  Reciprocal regulation of Δ1-pyrroline-5-carboxylate synthetase and proline dehydrogenase genes controls proline levels during and after osmotic stress in plants , 1996, Molecular and General Genetics MGG.

[37]  R. E. Sharp,et al.  Growth of the Maize Primary Root at Low Water Potentials : III. Role of Increased Proline Deposition in Osmotic Adjustment. , 1991, Plant physiology.

[38]  I. D. Teare,et al.  Rapid determination of free proline for water-stress studies , 1973, Plant and Soil.

[39]  A. Hoekema,et al.  A small-scale procedure for the rapid isolation of plant RNAs. , 1989, Nucleic acids research.

[40]  Z. Hong,et al.  Overexpression of [delta]-Pyrroline-5-Carboxylate Synthetase Increases Proline Production and Confers Osmotolerance in Transgenic Plants , 1995, Plant physiology.

[41]  E. T. Palva,et al.  Drought tolerance in tobacco , 1996, Nature.

[42]  Jian-Kang Zhu,et al.  Genetic Analysis of Salt Tolerance in Arabidopsis: Evidence for a Critical Role of Potassium Nutrition , 1998, Plant Cell.

[43]  M. Ishitani,et al.  Synechococcus sp. PCC7942 Transformed with Escherichia coli bet Genes Produces Glycine Betaine from Choline and Acquires Resistance to Salt Stress , 1995, Plant physiology.

[44]  D. Rudulier,et al.  Effects of Salt Stress on Amino Acid, Organic Acid, and Carbohydrate Composition of Roots, Bacteroids, and Cytosol of Alfalfa (Medicago sativa L.). , 1991, Plant physiology.

[45]  J. A. Hellebust,et al.  The regulation of amino acid metabolism during hyperosmotic stress in Acanthamoeba castellanii , 1987 .

[46]  F. Skoog,et al.  A revised medium for rapid growth and bio assays with tobacco tissue cultures , 1962 .

[47]  C. Kao,et al.  Characteristics of the Induction of the Accumulation of Proline by Abscisic Acid and Isobutyric Acid in Detached Rice Leaves , 1991 .

[48]  C. Koncz,et al.  Isolation and characterization of two different cDNAs of Δ1-pyrroline-5-carboxylate synthase in alfalfa, transcriptionally induced upon salt stress , 1998, Plant Molecular Biology.

[49]  B. V. Reddy,et al.  Proline-protein interactions: protection of structural and functional integrity of M4 lactate dehydrogenase. , 1994, Biochemical and biophysical research communications.

[50]  N. Buhot,et al.  Transcriptional regulation of proline biosynthesis in Medicago truncatula reveals developmental and environmental specific features. , 2004, Physiologia plantarum.