Deficiency in plastidic glutamine synthetase alters proline metabolism and transcriptomic response in Lotus japonicus under drought stress.

The role of plastidic glutamine synthetase (GS2) in proline biosynthesis and drought stress responses in Lotus japonicus was investigated using the GS2 mutant, Ljgln2-2. Wild-type (WT) and mutant plants were submitted to different lengths of time of water and nutrient solution deprivation. Several biochemical markers were measured and the transcriptional response to drought was determined by both quantitative real-time polymerase chain reaction and transcriptomics. The Ljgln2-2 mutant exhibited normal sensitivity to mild water deprivation, but physiological, biochemical and massive transcriptional differences were detected in the mutant, which compromised recovery (rehydration) following re-watering after severe drought stress. Proline accumulation during drought was substantially lower in mutant than in WT plants, and significant differences in the pattern of expression of the genes involved in proline metabolism were observed. Transcriptomic analysis revealed that about three times as many genes were regulated in response to drought in Ljgln2-2 plants compared with WT. The transcriptomic and accompanying biochemical data indicate that the Ljgln2-2 mutant is subject to more intense cellular stress than WT during drought. The results presented here implicate plastidic GS2 in proline production during stress and provide interesting insights into the function of proline in response to drought.

[1]  M. Udvardi,et al.  Mining for robust transcriptional and metabolic responses to long-term salt stress: a case study on the model legume Lotus japonicus. , 2010, Plant, cell & environment.

[2]  A. Savouré,et al.  Proline: a multifunctional amino acid. , 2010, Trends in plant science.

[3]  Michael K. Udvardi,et al.  Dissection of Symbiosis and Organ Development by Integrated Transcriptome Analysis of Lotus japonicus Mutant and Wild-Type Plants , 2009, PloS one.

[4]  M. Delledonne,et al.  Symbiotic competence in Lotus japonicus is affected by plant nitrogen status: transcriptomic identification of genes affected by a new signalling pathway. , 2009, The New phytologist.

[5]  S. Bernard,et al.  The importance of cytosolic glutamine synthetase in nitrogen assimilation and recycling. , 2009, The New phytologist.

[6]  Matthew Hannah,et al.  Genome-wide reprogramming of regulatory networks, transport, cell wall and membrane biogenesis during arbuscular mycorrhizal symbiosis in Lotus japonicus. , 2009, The New phytologist.

[7]  N. Saibo,et al.  Transcription Factors and Regulation of Photosynthetic and Related Metabolism under Environmental Stresses , 2022 .

[8]  Aihua Liu,et al.  Proline accumulation and transcriptional regulation of proline biosynthesis and degradation in Brassica napus. , 2009, BMB reports.

[9]  S. Binder,et al.  A nuclear gene encoding mitochondrial Delta-pyrroline-5-carboxylate dehydrogenase and its potential role in protection from proline toxicity. , 2008, The Plant journal : for cell and molecular biology.

[10]  D. Funck,et al.  Ornithine-δ-aminotransferase is essential for Arginine Catabolism but not for Proline Biosynthesis , 2008, BMC Plant Biology.

[11]  Christian Hermans,et al.  Proline accumulation in plants: a review , 2008, Amino Acids.

[12]  Henning Redestig,et al.  Integrative functional genomics of salt acclimatization in the model legume Lotus japonicus. , 2007, The Plant journal : for cell and molecular biology.

[13]  Yongze Yuan,et al.  Glutamine synthetase and glutamate dehydrogenase contribute differentially to proline accumulation in leaves of wheat (Triticum aestivum) seedlings exposed to different salinity. , 2007, Journal of plant physiology.

[14]  T. Arcondéguy,et al.  Molecular analysis of two mutants from Lotus japonicus deficient in plastidic glutamine synthetase: functional properties of purified GLN2 enzymes , 2006, Planta.

[15]  J. Monza,et al.  Nitrate assimilation in Lotus japonicus. , 2005, Journal of experimental botany.

[16]  O. Borsani,et al.  Osmotically Induced Proline Accumulation in Lotus Corniculatus Leaves is Affected by Light and Nitrogen Source , 2005, Plant Growth Regulation.

[17]  Joachim Selbig,et al.  Extension of the Visualization Tool MapMan to Allow Statistical Analysis of Arrays, Display of Coresponding Genes, and Comparison with Known Responses1 , 2005, Plant Physiology.

[18]  S. Tabata,et al.  Lotus japonicus: legume research in the fast lane. , 2005, Trends in plant science.

[19]  J. Teixeira,et al.  Regulation of glutamine synthetase expression in sunflower cells exposed to salt and osmotic stress , 2004 .

[20]  E. Pajuelo,et al.  Isolation of photorespiratory mutants from Lotus japonicus deficient in glutamine synthetase. , 2002, Physiologia plantarum.

[21]  E. Pajuelo,et al.  Characterisation and expression studies of a root cDNA encoding for ferredoxin-nitrite reductase from Lotus japonicus. , 2001, Physiologia plantarum.

[22]  O. Borsani,et al.  Water stress generates an oxidative stress through the induction of a specific Cu/Zn superoxide dismutase in Lotus corniculatus leaves , 2001 .

[23]  H. Hoshida,et al.  Enhanced tolerance to salt stress in transgenic rice that overexpresses chloroplast glutamine synthetase , 2000, Plant Molecular Biology.

[24]  Y. Roux,et al.  Glutamine Synthetase in the Phloem Plays a Major Role in Controlling Proline Production , 1999, Plant Cell.

[25]  O. Borsani,et al.  Proline is involved in water stress responses of Lotus corniculatus nitrogen fixing and nitrate fed plants , 1999 .

[26]  S. Mehta,et al.  Heavy‐metal‐induced proline accumulation and its role in ameliorating metal toxicity in Chlorella vulgaris , 1999 .

[27]  S. Lutts,et al.  NaCl effects on proline metabolism in rice (Oryza sativa) seedlings , 1999 .

[28]  N. Roosens,et al.  Isolation of the ornithine-delta-aminotransferase cDNA and effect of salt stress on its expression in Arabidopsis thaliana. , 1998, Plant physiology.

[29]  B. Forde,et al.  Regulation of the expression of ferredoxin-glutamate synthase in barley , 1997, Planta.

[30]  P. D. Hare,et al.  Metabolic implications of stress-induced proline accumulation in plants , 1997, Plant Growth Regulation.

[31]  A. Migge,et al.  A role for cytosolic glutamine synthetase in the remobilization of leaf nitrogen during water stress in tomato , 1997 .

[32]  A. Kozaki,et al.  Photorespiration protects C3 plants from photooxidation , 1996, Nature.

[33]  K. Yamaguchi-Shinozaki,et al.  A nuclear gene encoding mitochondrial proline dehydrogenase, an enzyme involved in proline metabolism, is upregulated by proline but downregulated by dehydration in Arabidopsis. , 1996, The Plant cell.

[34]  V. Valpuesta,et al.  Salt stress increases ferredoxin-dependent glutamate synthase activity and protein level in the leaves of tomato , 1995 .

[35]  E. Bray Molecular Responses to Water Deficit , 1993, Plant physiology.

[36]  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.

[37]  J. Stougaard,et al.  Lotus japonicus, an autogamous, diploid legume species for classical and molecular genetics , 1992 .

[38]  E. Ábrahám,et al.  Duplicated P5CS genes of Arabidopsis play distinct roles in stress regulation and developmental control of proline biosynthesis. , 2008, The Plant journal : for cell and molecular biology.

[39]  A. J. Márquez Lotus japonicus handbook , 2005 .

[40]  Suresh Iyer,et al.  Products of Proline Catabolism Can Induce Osmotically Regulated Genes in Rice , 1998 .