Enhancing Heat Tolerance of the Little Dogwood Cornus canadensis L. f. with Introduction of a Superoxide Reductase Gene from the Hyperthermophilic Archaeon Pyrococcus furiosus

Production of reactive oxygen species (ROS) can be accelerated under various biotic and abiotic stresses causing lipid peroxidation, protein degradation, enzyme inactivation, and DNA damage. Superoxide reductase (SOR) is a novel antioxidant enzyme from Pyrococcus furiosus and is employed by this anaerobic hyperthermophilic archaeon for efficient detoxification of ROS. In this study, SOR was introduced into a flowering plant Cornus canadensis to enhance its heat tolerance and reduce heat induced damage. A fusion construct of the SOR gene and Green Fluorescent Protein gene (GFP) was introduced into C. canadensis using Agrobacterium-mediated transformation. Heat tolerance of the GFP-SOR expressing transgenic plants was investigated by observing morphological symptoms of heat injury and by examining changes in photosynthesis, malondialdehyde (MDA), and proline levels in the plants. Our results indicate that the expression of the P. furiosus SOR gene in the transgenic plants alleviated lipid peroxidation of cell membranes and photoinhibition of PS II, and decreased the accumulation of proline at 40°C. After a series of exposures to increasing temperatures, the SOR transgenic plants remained healthy and green whereas most of the non-transgenic plants dried up and were unable to recover. While it had previously been reported that expression of SOR in Arabidopsis enhanced heat tolerance, this is the first report of the successful demonstration of improved heat tolerance in a non-model plant resulting from the introduction of P. furiosus SOR. The study demonstrates the potential of SOR for crop improvement and that inherent limitations of plant heat tolerance can be ameliorated with P. furiosus SOR.

[1]  P. Sears,et al.  superoxide dismutase , 2020, Catalysis from A to Z.

[2]  C. Abdelly,et al.  How reactive oxygen species and proline face stress together. , 2014, Plant physiology and biochemistry : PPB.

[3]  N. Suzuki,et al.  ROS as key players in plant stress signalling. , 2014, Journal of experimental botany.

[4]  O. Matsuda,et al.  Thermotolerant cyclamen with reduced acrolein and methyl vinyl ketone , 2012, Journal of experimental botany.

[5]  N. Suzuki,et al.  ROS and redox signalling in the response of plants to abiotic stress. , 2012, Plant, cell & environment.

[6]  H. Nayyar,et al.  Proline induces heat tolerance in chickpea (Cicer arietinum L.) plants by protecting vital enzymes of carbon and antioxidative metabolism , 2011, Physiology and Molecular Biology of Plants.

[7]  M. Zhang,et al.  Proline Accumulation Is Inhibitory to Arabidopsis Seedlings during Heat Stress1[W] , 2011, Plant Physiology.

[8]  K. Vandepoele,et al.  ROS signaling: the new wave? , 2011, Trends in plant science.

[9]  H. Nayyar,et al.  Heat-stress induced inhibition in growth and chlorosis in mungbean (Phaseolus aureus Roxb.) is partly mitigated by ascorbic acid application and is related to reduction in oxidative stress , 2011, Acta Physiologiae Plantarum.

[10]  M. Farooq,et al.  Role of proline and glycinebetaine pretreatments in improving heat tolerance of sprouting sugarcane (Saccharum sp.) buds , 2011, Plant Growth Regulation.

[11]  D. Yun,et al.  Enhanced tolerance to methyl viologen-induced oxidative stress and high temperature in transgenic potato plants overexpressing the CuZnSOD, APX and NDPK2 genes. , 2010, Physiologia plantarum.

[12]  Md. Anamul Hoque,et al.  Exogenous proline and glycinebetaine increase antioxidant enzyme activities and confer tolerance to cadmium stress in cultured tobacco cells. , 2009, Journal of plant physiology.

[13]  A. Grunden,et al.  Expression of Pyrococcus furiosus Superoxide Reductase in Arabidopsis Enhances Heat Tolerance1[C][W][OA] , 2009, Plant Physiology.

[14]  H. Okubo,et al.  Effects of Cold Storage and Different Pulsing Treatments on Postharvest Quality of Cut OT Lily 'Mantissa' Flowers , 2009 .

[15]  R. Qu,et al.  Shoot regeneration of dwarf dogwood (Cornus canadensis L.) and morphological characterization of the regenerated plants , 2009, Plant Cell, Tissue and Organ Culture (PCTOC).

[16]  Norio Murata,et al.  How do environmental stresses accelerate photoinhibition? , 2008, Trends in plant science.

[17]  M. H. D. Carvalho.  Drought stress and reactive oxygen species , 2008 .

[18]  E. Farmer,et al.  Reactive electrophile species. , 2007, Current opinion in plant biology.

[19]  S. Allakhverdiev,et al.  Photoinhibition of photosystem II under environmental stress. , 2007, Biochimica et biophysica acta.

[20]  Andreas Hansson,et al.  Oxidative modifications to cellular components in plants. , 2007, Annual review of plant biology.

[21]  A. Wahid,et al.  Expression of dehydrins under heat stress and their relationship with water relations of sugarcane leaves , 2007, Biologia Plantarum.

[22]  E. Stephanou,et al.  Abiotic Stress Generates ROS That Signal Expression of Anionic Glutamate Dehydrogenases to Form Glutamate for Proline Synthesis in Tobacco and Grapevine[W] , 2006, The Plant Cell Online.

[23]  R. Sairam,et al.  Protective role of antioxidant enzymes under high temperature stress. , 2006, Plant science : an international journal of experimental plant biology.

[24]  Yanping Guo,et al.  Photosynthetic characteristics and protective mechanisms against photooxidation during high temperature stress in two citrus species , 2006 .

[25]  Juan José Alarcón,et al.  Changes in photosynthetic parameters and antioxidant activities following heat-shock treatment in tomato plants. , 2006, Functional plant biology : FPB.

[26]  A. Grunden,et al.  Production of a thermostable archaeal superoxide reductase in plant cells , 2005, FEBS letters.

[27]  M. Adams,et al.  In Vitro Reconstitution of an NADPH-Dependent Superoxide Reduction Pathway from Pyrococcus furiosus , 2005, Applied and Environmental Microbiology.

[28]  R. Mittler,et al.  Reactive oxygen gene network of plants. , 2004, Trends in plant science.

[29]  Stephen M. Schrader,et al.  Electron transport is the functional limitation of photosynthesis in field-grown Pima cotton plants at high temperature , 2004 .

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

[31]  E. Marra,et al.  Production of Reactive Oxygen Species, Alteration of Cytosolic Ascorbate Peroxidase, and Impairment of Mitochondrial Metabolism Are Early Events in Heat Shock-Induced Programmed Cell Death in Tobacco Bright-Yellow 2 Cells1 , 2004, Plant Physiology.

[32]  R. Volkov,et al.  Heat Stress- and Heat Shock Transcription Factor-Dependent Expression and Activity of Ascorbate Peroxidase in Arabidopsis1 , 2002, Plant Physiology.

[33]  M. Knight,et al.  Protection against Heat Stress-Induced Oxidative Damage in Arabidopsis Involves Calcium, Abscisic Acid, Ethylene, and Salicylic Acid , 2002, Plant Physiology.

[34]  K. Jones,et al.  Iron-superoxide dismutase expression in transgenic alfalfa increases winter survival without a detectable increase in photosynthetic oxidative stress tolerance. , 2000, Plant physiology.

[35]  Bingru Huang,et al.  Heat Stress Injury in Relation to Membrane Lipid Peroxidation in Creeping Bentgrass , 2000 .

[36]  M. Adams,et al.  Anaerobic microbes: oxygen detoxification without superoxide dismutase. , 1999, Science.

[37]  W. Van Camp,et al.  Enhancement of Oxidative Stress Tolerance in Transgenic Tobacco Plants Overproducing Fe-Superoxide Dismutase in Chloroplasts , 1996, Plant physiology.

[38]  M. Yamada,et al.  Heat tolerance in leaves of tropical fruit crops as measured by chlorophyll fluorescence , 1996 .

[39]  K Botzenhart,et al.  Reactive Oxygen Species , 2014 .

[40]  R. Allen,et al.  Dissection of Oxidative Stress Tolerance Using Transgenic Plants , 1995, Plant physiology.

[41]  J. Sherwood,et al.  Enhanced recovery of transformants of Agrobacterium tumefaciens after freeze-thaw transformation and drug selection. , 1994, BioTechniques.

[42]  J. Cairney,et al.  A simple and efficient method for isolating RNA from pine trees , 1993, Plant Molecular Biology Reporter.

[43]  J. Berry,et al.  Photosynthetic Response and Adaptation to Temperature in Higher Plants , 1980 .

[44]  R. Qu,et al.  Plant regeneration and genetic transformation of C. canadensis: a non-model plant appropriate for investigation of flower development in Cornus (Cornaceae) , 2012, Plant Cell Reports.

[45]  N. Babu.  High temperature and salt stress response in French bean ( Phaseolus vulgaris ) , 2008 .

[46]  M. H. Cruz de Carvalho.  Drought stress and reactive oxygen species: Production, scavenging and signaling. , 2008, Plant signaling & behavior.

[47]  R. Nageshbabu,et al.  High temperature and salt stress response in French bean ( Phaseolus vulgaris ) , 2008 .

[48]  P. B. Allen,et al.  Electron transport in , 2006 .

[49]  N. Sreenivasulu,et al.  Regulation of proline biosynthesis, degradation, uptake and transport in higher plants: Its implications in plant growth and abiotic stress tolerance , 2005 .

[50]  Zhong-Guang Li,et al.  Heat-shock-induced cross adaptation to heat, chilling, drought and salt stress in maize seedlings and involvement of H2O2 , 2001 .

[51]  Dirk Inzé,et al.  SUPEROXIDE DISMUTASE AND STRESS TOLERANCE , 1992 .