An Arabidopsis Mitochondrial Uncoupling Protein Confers Tolerance to Drought and Salt Stress in Transgenic Tobacco Plants

Background Plants are challenged by a large number of environmental stresses that reduce productivity and even cause death. Both chloroplasts and mitochondria produce reactive oxygen species under normal conditions; however, stress causes an imbalance in these species that leads to deviations from normal cellular conditions and a variety of toxic effects. Mitochondria have uncoupling proteins (UCPs) that uncouple electron transport from ATP synthesis. There is evidence that UCPs play a role in alleviating stress caused by reactive oxygen species overproduction. However, direct evidence that UCPs protect plants from abiotic stress is lacking. Methodology/Principal Findings Tolerances to salt and water deficit were analyzed in transgenic tobacco plants that overexpress a UCP (AtUCP1) from Arabidopsis thaliana. Seeds of AtUCP1 transgenic lines germinated faster, and adult plants showed better responses to drought and salt stress than wild-type (WT) plants. These phenotypes correlated with increased water retention and higher gas exchange parameters in transgenic plants that overexpress AtUCP1. WT plants exhibited increased respiration under stress, while transgenic plants were only slightly affected. Furthermore, the transgenic plants showed reduced accumulation of hydrogen peroxide in stressed leaves compared with WT plants. Conclusions/Significance Higher levels of AtUCP1 improved tolerance to multiple abiotic stresses, and this protection was correlated with lower oxidative stress. Our data support previous assumptions that UCPs reduce the imbalance of reactive oxygen species. Our data also suggest that UCPs may play a role in stomatal closure, which agrees with other evidence of a direct relationship between these proteins and photosynthesis. Manipulation of the UCP protein expression in mitochondria is a new avenue for crop improvement and may lead to crops with greater tolerance for challenging environmental conditions.

[1]  E. Ratajczak,et al.  Production and scavenging of reactive oxygen species in Fagus sylvatica seeds during storage at varied temperature and humidity. , 2005, Journal of plant physiology.

[2]  I. Møller PLANT MITOCHONDRIA AND OXIDATIVE STRESS: Electron Transport, NADPH Turnover, and Metabolism of Reactive Oxygen Species. , 2001, Annual review of plant physiology and plant molecular biology.

[3]  F. Loreto,et al.  12CO2 emission from different metabolic pathways measured in illuminated and darkened C3 and C4 leaves at low, atmospheric and elevated CO2 concentration. , 2003, Journal of experimental botany.

[4]  L. Mcintosh,et al.  The alternative oxidase lowers mitochondrial reactive oxygen production in plant cells. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[5]  J. Riesmeier,et al.  A plant cold-induced uncoupling protein , 1997, Nature.

[6]  W. Emmerich,et al.  Effect of polyethylene glycol exclusion on the water potential of solution-saturated filter paper. , 1990, Plant physiology.

[7]  T. Dučić,et al.  Activities of Antioxidant Systems During Germination of Chenopodium rubrum Seeds , 2003, Biologia Plantarum.

[8]  F. Fiorani,et al.  Characterization of Transformed Arabidopsis with Altered Alternative Oxidase Levels and Analysis of Effects on Reactive Oxygen Species in Tissue1[W] , 2005, Plant Physiology.

[9]  A. Zachowski,et al.  The plant uncoupling protein homologues: a new family of energy-dissipating proteins in plant mitochondria. , 2004, Plant physiology and biochemistry : PPB.

[10]  Kikukatsu Ito Isolation of two distinct cold-inducible cDNAs encoding plant uncoupling proteins from the spadix of skunk cabbage (Symplocarpus foetidus) , 1999 .

[11]  A. Rasmusson,et al.  Light Regulation of the Arabidopsis Respiratory Chain. Multiple Discrete Photoreceptor Responses Contribute to Induction of Type II NAD(P)H Dehydrogenase Genes1 , 2004, Plant Physiology.

[12]  J. Zhu,et al.  Cell signaling under salt, water and cold stresses. , 2001, Current opinion in plant biology.

[13]  J. Flexas,et al.  Keeping a positive carbon balance under adverse conditions: responses of photosynthesis and respiration to water stress , 2006 .

[14]  A. Vercesi,et al.  Overexpression of Plant Uncoupling Mitochondrial Protein in Transgenic Tobacco Increases Tolerance to Oxidative Stress , 2003, Journal of bioenergetics and biomembranes.

[15]  M. Tester,et al.  Mechanisms of salinity tolerance. , 2008, Annual review of plant biology.

[16]  H. Hirt,et al.  Reactive oxygen species: metabolism, oxidative stress, and signal transduction. , 2004, Annual review of plant biology.

[17]  A. Altman,et al.  Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance , 2003, Planta.

[18]  K. Thompson,et al.  Seeds: Physiology of Development and Germination , 1986 .

[19]  O. Atkin,et al.  Transient shade and drought have divergent impacts on the temperature sensitivity of dark respiration in leaves of Geum urbanum. , 2008, Functional plant biology : FPB.

[20]  Alain Vavasseur,et al.  Arabidopsis OST1 Protein Kinase Mediates the Regulation of Stomatal Aperture by Abscisic Acid and Acts Upstream of Reactive Oxygen Species Production Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.007906. , 2002, The Plant Cell Online.

[21]  C. Sluse-Goffart,et al.  Mitochondrial UCPs: new insights into regulation and impact. , 2006, Biochimica et biophysica acta.

[22]  P. Ježek,et al.  On the Mechanism of Fatty Acid-induced Proton Transport by Mitochondrial Uncoupling Protein (*) , 1996, The Journal of Biological Chemistry.

[23]  J. Gebicki,et al.  Hydroperoxide assay with the ferric-xylenol orange complex. , 1999, Analytical biochemistry.

[24]  C. Bailly Active oxygen species and antioxidants in seed biology , 2004, Seed Science Research.

[25]  J. Flexas,et al.  Photosynthesis under drought and salt stress: regulation mechanisms from whole plant to cell. , 2009, Annals of botany.

[26]  M. Loureiro,et al.  Drought tolerance is associated with rooting depth and stomatal control of water use in clones of Coffea canephora. , 2005, Annals of botany.

[27]  J. Hancock,et al.  Regulation of the Arabidopsis transcriptome by oxidative stress. , 2001, Plant physiology.

[28]  Hur-Song Chang,et al.  Transcriptome Changes for Arabidopsis in Response to Salt, Osmotic, and Cold Stress1,212 , 2002, Plant Physiology.

[29]  A. Vercesi,et al.  PUMPing plants , 1995, Nature.

[30]  J. Guiamet,et al.  Up-regulation of the mitochondrial alternative oxidase pathway enhances photosynthetic electron transport under drought conditions. , 2005, Journal of experimental botany.

[31]  L. Sweetlove,et al.  Activation and Function of Mitochondrial Uncoupling Protein in Plants* , 2004, Journal of Biological Chemistry.

[32]  N. Di Fonzo,et al.  Possible Plant Mitochondria Involvement in Cell Adaptation to Drought Stress a Case Study: Durum Wheat Mitochondria , 2006 .

[33]  Alisdair R Fernie,et al.  Mitochondrial uncoupling protein is required for efficient photosynthesis , 2006, Proceedings of the National Academy of Sciences.

[34]  P. Mullineaux,et al.  Tolerance of pea (Pisum sativum L.) to long‐term salt stress is associated with induction of antioxidant defences , 2000 .

[35]  Tracey Ann Cuin,et al.  Plant Salt Tolerance , 2012, Methods in Molecular Biology.

[36]  D. Nicholls,et al.  Brown-adipose-tissue mitochondria: photoaffinity labelling of the regulatory site of energy dissipation. , 1978, European journal of biochemistry.

[37]  K. Shinozaki,et al.  Molecular responses to dehydration and low temperature: differences and cross-talk between two stress signaling pathways. , 2000, Current opinion in plant biology.

[38]  J. W. Outlaw Integration of Cellular and Physiological Functions of Guard Cells , 2003 .

[39]  Nobuhiro Suzuki,et al.  Reactive oxygen species homeostasis and signalling during drought and salinity stresses. , 2010, Plant, cell & environment.

[40]  Yves Gibon,et al.  Deficiency of mitochondrial fumarase activity in tomato plants impairs photosynthesis via an effect on stomatal function. , 2007, The Plant journal : for cell and molecular biology.

[41]  K. Akiyama,et al.  Monitoring the expression profiles of 7000 Arabidopsis genes under drought, cold and high-salinity stresses using a full-length cDNA microarray. , 2002, The Plant journal : for cell and molecular biology.

[42]  Q. Gao,et al.  Drought tolerance through overexpression of monoubiquitin in transgenic tobacco. , 2008, Journal of plant physiology.

[43]  N. Di Fonzo,et al.  Reactive oxygen species inhibit the succinate oxidation‐supported generation of membrane potential in wheat mitochondria , 2002, FEBS letters.

[44]  W. Sutherland,et al.  Reaping the Benefits: Science and the sustainable intensification of global agriculture , 2009 .

[45]  A. Vercesi,et al.  Plant uncoupling mitochondrial proteins. , 2006, Annual review of plant biology.

[46]  Jia Chen,et al.  Transgenic Arabidopsis overexpressing Mn-SOD enhanced salt-tolerance , 2004 .

[47]  T. Takano,et al.  Two cysteine proteinase inhibitors from Arabidopsis thaliana, AtCYSa and AtCYSb, increasing the salt, drought, oxidation and cold tolerance , 2008, Plant Molecular Biology.

[48]  V. Mittova,et al.  Salinity up-regulates the antioxidative system in root mitochondria and peroxisomes of the wild salt-tolerant tomato species Lycopersicon pennellii. , 2004, Journal of experimental botany.

[49]  J. Schroeder,et al.  GUARD CELL SIGNAL TRANSDUCTION. , 2003, Annual review of plant physiology and plant molecular biology.

[50]  D. Macherel,et al.  The crucial role of plant mitochondria in orchestrating drought tolerance. , 2009, Annals of botany.

[51]  L. Sweetlove,et al.  Superoxide Stimulates a Proton Leak in Potato Mitochondria That Is Related to the Activity of Uncoupling Protein* , 2003, Journal of Biological Chemistry.

[52]  A. Vercesi,et al.  ZmPUMP encodes a maize mitochondrial uncoupling protein that is induced by oxidative stress , 2003 .

[53]  A. Vercesi,et al.  The plant energy-dissipating mitochondrial systems: depicting the genomic structure and the expression profiles of the gene families of uncoupling protein and alternative oxidase in monocots and dicots. , 2006, Journal of experimental botany.

[54]  D. Nicholls,et al.  A History of the First Uncoupling Protein, UCP1 , 1999, Journal of bioenergetics and biomembranes.

[55]  M. M. Chaves,et al.  Photosynthesis and drought: can we make metabolic connections from available data? , 2011, Journal of experimental botany.

[56]  Jian-Kang Zhu,et al.  Salt and drought stress signal transduction in plants. , 2002, Annual review of plant biology.

[57]  A. S. Raghavendra,et al.  Importance of ROS and antioxidant system during the beneficial interactions of mitochondrial metabolism with photosynthetic carbon assimilation , 2009, Planta.

[58]  J. Borecký,et al.  Mitochondrial Uncoupling Proteins in Mammals and Plants , 2001, Bioscience reports.

[59]  Stan D. Wullschleger,et al.  A review of whole-plant water use studies in tree. , 1998, Tree physiology.