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Two rice cytosolic ascorbate peroxidases differentially improve salt tolerance in transgenic Arabidopsis

  • Biotic and Abiotic Stress
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Abstract

In order to determine the different roles of rice (Oryza sativa L.) cytosolic ascorbate peroxidases (OsAPXa and OsAPXb, GenBank accession nos. D45423 and AB053297, respectively) under salt stress, transgenic Arabidopsis plants over-expressing OsAPXa or OsAPXb were generated, and they all exhibited increased tolerance to salt stress compared to wild-type plants. Moreover, transgenic lines over-expressing OsAPXb showed higher salt tolerance than OsAPXa transgenic lines as indicated by root length and total chlorophyll content. In addition to ascorbate peroxidase (APX) activity, antioxidant enzyme activities of catalase (CAT), superoxide dismutase (SOD) and glutathione reductase (GR), which are also involved in the salt tolerance process, and the content of H2O2 were also assayed in both transgenic and wild-type plants. The results showed that the overproduction of OsAPXb enhanced and maintained APX activity to a much higher degree than OsAPXa in transgenic Arabidopsis during treatment with different concentrations of NaCl, enhanced the active oxygen scavenging system, and protected plants from salt stress by equilibrating H2O2 metabolism. Our findings suggest that the rice cytosolic OsAPXb gene has a more functional role than OsAPXa in the improvement of salt tolerance in transgenic plants.

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Abbreviations

APX:

Ascorbate peroxidase

cAPX:

Cytosolic ascorbate peroxidase

CAT:

Catalase

GR:

Glutathione reductase

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

References

  • Abei H (1974) Catalases. In: Bergmeyer HU (eds) Methods of enzymatic analysis. vol. 2, Academic, New York, pp 673–684

    Google Scholar 

  • Arnon DI (1949) Copper enzymes in isolated chloroplasts in Beta vulgaris. Plant Physiol 24:1–15

    PubMed  CAS  Google Scholar 

  • Badawi GH, Kawano N, Yamauchi Y, Shimada E, Sasaki R, Kubo A, Tanaka K (2004) Over-expression of ascorbate peroxidase in tobacco chloroplasts enhances the tolerance to salt stress and water deficit. Physiol Plant 121:231–238

    Article  PubMed  CAS  Google Scholar 

  • Bernt E, Bergmeyer HU (1974) Inorganic peroxides. In: Bergmeyer HU (ed) Methods of enzymatic analysis, vol. 4, Academic, New York, pp 2246–2248

    Google Scholar 

  • Bradford M (1976) A rapid and sensitive method for the quantification of microgram quantities of proteins utilizing the principal of protein–dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  • Chen GX, Asada K (1989) Ascorbate peroxidase in tea leaves: occurrence of two isozymes and the differences in their enzymatic and molecular properties. Plant Cell Physiol 30:987–998

    CAS  Google Scholar 

  • Davletova S, Rizhsky L, Liang H, Shengqiang Z, Oliver DJ, Coutu J, Shulaev V, Schlauch K, Mittler R (2005) Cytosolic ascorbate peroxidase 1 is a central component of the reactive oxygen gene network of Arabidopsis. Plant Cell 17:268–281

    Article  PubMed  CAS  Google Scholar 

  • Feierabend J, Dehne S (1996) Fate of the porphyrin cofactors during the light-dependent turnover of catalase and of the photosystem II reaction-center protein D1 in mature rye leaves. Planta 198:413–422

    Article  CAS  Google Scholar 

  • Fourcroy P, Vansuyt G, Kushnir S, Inzé D, Briat J (2004) Iron-regulated expression of a cytosolic ascorbate peroxidase encoded by the APX1 gene in Arabidopsis seedlings. Plant Physiol 134:605–613

    Article  PubMed  CAS  Google Scholar 

  • Foyer CH, Lelandais M, Kenert KJ (1994) Photooxidative stress in plants. Physiol Plant 92:696–717

    Article  CAS  Google Scholar 

  • Giannopolitis CN, Ries SK (1977) Superoxide dismutases. I. Occurrence in higher plants. Plant Physiol 59:309–314

    Article  PubMed  CAS  Google Scholar 

  • Harlow E, Lane D (1988) Antibodies: a laboratory manual. Cold Spring Harbor Lab Press, Plainview

    Google Scholar 

  • Ishikawa T, Madhusudhan R, Shigeoka S (2003) Effects of iron on the expression of ascorbate peroxidase in Euglena gracilis. Plant Sci 165:1363–1367

    Article  CAS  Google Scholar 

  • Lee DH, Lee CB (2000) Chilling stress-induced changes of antioxidant enzymes in the leaves of cucumber: in gel enzyme activity assay. Plant Sci 159:75–85

    Article  PubMed  CAS  Google Scholar 

  • Lee DH, Kim YS, Lee CB (2001) The inductive responses of the antioxidant enzymes by salt stress in the rice (Oryza sativa L.). J Plant Physiol 158:737–745

    Article  CAS  Google Scholar 

  • Lu ZQ, Takano T, Liu SK (2005) Purification and characterization of two ascorbate peroxidases of rice (Oryza sativa L.) expressed in Escherichia coli. Biotechnol Lett 27:63–67

    Article  PubMed  CAS  Google Scholar 

  • Menezes-Benavente L, Teixeira FK, Kamei CLA, Margis-Pinheiro M (2004) Salt stress induces altered expression of genes encoding antioxidant enzymes in seedlings of a Brazilian indica rice (Oryza sativa L.). Plant Sci 166:323–331

    Article  CAS  Google Scholar 

  • Mittler R, Zilinskas BA (1994) Regulation of pea cytosolic ascorbate peroxidase and other antioxidant enzymes during the progression of drought stress and following recovery from drought. Plant J 5(3):397–405

    Article  PubMed  CAS  Google Scholar 

  • Orendi G, Zimmermann P, Baar C, Zentgraf U (2001) Loss of stress-induced expression of catalases 3 during leaf senescence in Arabidopsis thaliana is restricted to oxidative stress. Plant Sci 161:301–314

    Article  PubMed  CAS  Google Scholar 

  • Panchuk II, Volkov RA, Schöffl F (2002) Heat stress- and heat shock transcription factor-dependent expression and activity of ascorbate peroxidase in Arabidopsis. Plant Physiol 129:838–853

    Article  PubMed  CAS  Google Scholar 

  • Panchuk II, Zentgraf U, Volkov RA (2005) Expression of the Apx gene family during leaf senescence of Arabidopsis thaliana. Planta 222:926–932

    Article  PubMed  CAS  Google Scholar 

  • Polle A (1997) Defense against photooxidative damage in plants. In: Scandalios J (ed) Oxidative stress and molecular biology of antioxidant defenses. Cold Spring Harbor Laboratory Press. Woodbury, pp 785–813

    Google Scholar 

  • Rao MV, Paliyath G, Ormrod DP (1996) Ultraviolet-B- and ozone-induced biochemical changes in antioxidant enzymes of Arabidopsis thaliana. Plant Physiol 110:125–136

    Article  PubMed  CAS  Google Scholar 

  • Sano S, Ueda M, Kitajima S, Takeda T, Shigeoka S, Kurano N, Miyachi S, Miyake C, Yokota A (2001) Characterization of ascorbate peroxidases from unicellular red alga Galdieria partita. Plant Cell Physiol 42:433–440

    Article  PubMed  CAS  Google Scholar 

  • Sharma P, Dubey RS (2005) Drought induces oxidative stress and enhances the activities of antioxidant enzymes in growing rice seedlings. Plant Growth Regul 46:209–211

    Article  CAS  Google Scholar 

  • Shigeoka S, Ishikawa T, Tamoi M, Miyagawa Y, Takeda T, Yabuta Y, Yoshimura K (2002) Regulation and function of ascorbate peroxidase isoenzymes. J Exp Bot 53:1305–1319

    Article  PubMed  CAS  Google Scholar 

  • Shi H, Lee B, Wu SJ, Zhu JK (2002) Overexpression of a plasma membrane Na+/H+ antiporter gene improves salt tolerance in Arabidopsis thaliana. Nat Biotechnol 21:81–85

    Article  PubMed  CAS  Google Scholar 

  • Teixeira FK, Menezes-Benavente L, Galvão VC, Margis R, Margis-Pinheiro M (2006) Rice ascorbate peroxidase gene family encodes functionally diverse isoforms localized in different subcellular compartments. Planta 224:300–314

    Article  PubMed  CAS  Google Scholar 

  • Tsai YC, Hong CY, Liu LF, Kao CH (2005) Expression of ascorbate peroxidase and glutathione reductase in roots of rice seedlings in response to NaCl and H2O2. J Plant Physiol 162:291–299

    Article  PubMed  CAS  Google Scholar 

  • Wang J, Zhang H, Allen RD (1999) Overexpression of an Arabidopsis peroxisomal ascorbate peroxidase gene in tobacco increases protection against oxidative stress. Plant Cell Physiol 40:725–732

    PubMed  CAS  Google Scholar 

  • Wang WX, Vinocur B, Altman A (2003) A plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. Planta 218:1–14

    Article  PubMed  CAS  Google Scholar 

  • Wang Y, Wisniewski M, Meilan R, Cui M, Webb R, Fuchigami L (2005) Overexpression of cytosolic ascorbate peroxidase in tomato confers tolerance to chilling and salt stress. J Am Soc Hortic Sci 130:167–173

    CAS  Google Scholar 

  • Wehmeyer N, Hernandez LD, Finkelstein RR, Vierling E (1996) Synthesis of small heat-shock proteins is part of the developmental program of late seed maturation. Plant Physiol 112:747–757

    Article  PubMed  CAS  Google Scholar 

  • Zhu JK (2001) Plant salt tolerance. Trends Plant Sci 6:66–71

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Excellent Teachers Program Foundation of Helongjiang University and the National High Technology Research and Development Program (863 Program) from the People’s Republic of China.

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Authors and Affiliations

  1. Biochemistry and Molecular Biology Lab, College of Life Sciences, Heilongjiang University, Harbin, 150080, People’s Republic China

    Zhenqiang Lu

  2. Key Laboratory of Sugar Beet Genetics and Breeding, Academy of Crop Sciences, Heilongjiang University, Harbin, 150080, People’s Republic China

    Dali Liu

  3. Sugar Beet Research Institute, Chinese Academy of Agricultural Sciences, Harbin, 150080, People’s Republic China

    Dali Liu

  4. Alkali Soil Natural Environmental Science Center (ASNESC), Stress Molecular Biology Laboratory, Northeast Forestry University, Harbin, 150040, People’s Republic China

    Shenkui Liu

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  1. Zhenqiang Lu
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  2. Dali Liu
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  3. Shenkui Liu
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Corresponding author

Correspondence to Dali Liu.

Additional information

Communicated by W.-H. Wu.

Zhenqiang Lu and Dali Liu contributed equally.

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Lu, Z., Liu, D. & Liu, S. Two rice cytosolic ascorbate peroxidases differentially improve salt tolerance in transgenic Arabidopsis . Plant Cell Rep 26, 1909–1917 (2007). https://doi.org/10.1007/s00299-007-0395-7

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  • DOI: https://doi.org/10.1007/s00299-007-0395-7

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