Advertisement

Biologia Plantarum

, Volume 54, Issue 4, pp 614–620 | Cite as

Overexpression of tomato tAPX gene in tobacco improves tolerance to high or low temperature stress

  • W. -H. Sun
  • M. Duan
  • F. Li
  • D. -F. Shu
  • S. Yang
  • Q. -W. Meng
Original Papers

Abstract

In order to investigate the function of chloroplast ascorbate peroxidase under temperature stress, the thylakoid-bound ascorbate peroxidase gene from tomato leaf (TtAPX) was introduced into tobacco. Transformants were selected for their ability to grow on medium containing kanamycin. RNA gel blot analysis confirmed that TtAPX in tomato was induced by chilling or heat stress. Over-expression of TtAPX in tobacco improved seed germination under temperature stress. Two transgenic tobacco lines showed higher ascorbate peroxidase activity, accumulated less hydrogen peroxide and malondialdehyde than wild type plants under stress condition. The photochemical efficiency of photosystem 2 in the transgenic lines was distinctly higher than that of wild type plants under chilling and heat stresses. Results indicated that the over-expression of TtAPX enhanced tolerance to temperature stress in transgenic tobacco plants.

Additional key words

Lycopersicum esculentum malondialdehyde Nicotiana tabacum photosystem 2 thylakoid-bound ascorbate peroxidase transgenic tobacco 

Abbreviations

APX

ascorbate peroxidase

AsA

ascorbic acid

CAT

catalase

CK

control

F0

initial fluorescence

Fv/Fm

variable to maximum fluorescence ratio (maximum photochemical efficiency of PS 2)

MDA

malondialdehyde

O2·−

superoxide radical

PBS

phosphate-buffered saline

PN

net photosynthetic rate

PS 2

photosystem 2

REL

relative electrolyte leakage

ROS

reactive oxygen species

SDS

sodium dodecyl sulfate

SOD

superoxide dismutase

SSC

standard saline citrate

tAPX

thylakoid-bound ascorbate peroxidase

TBA

Tris-buffered acetate

TCA

trichloroacetic acid

tAPX

tomato thylakoid-bound ascorbate peroxidase gene

WT

wild type plant

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Notes

Acknowledgements

This research was supported by the State Key Basic Research and Development Plan of China (2009CB118500), the Natural Science Foundation of China (30871458) and Program for Changjiang Scholars and Innovative Research Team in University (Grant IRT0635).

References

  1. Asada, K.: Ascorbate peroxidase — a hydrogen peroxidescavenging enzyme in plants. — Physiol. Plant. 85: 235–241, 1992.CrossRefGoogle Scholar
  2. Asada, K.: The water-water cycle in chloroplasts: scavenging of active oxygens and dissipation of excess photons. — Annu. Rev. Plant Physiol. Plant mol. Biol. 50: 601–639, 1999.CrossRefPubMedGoogle Scholar
  3. Bhattacharjee, S.: Calcium-dependent signaling pathway in the heat-induced oxidative injury in Amaranthus lividus. — Biol. Plant. 52: 137–140, 2008.CrossRefGoogle Scholar
  4. Bradford, M.M.: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. — Anal. Biochem. 72: 248–254, 1976.CrossRefPubMedGoogle Scholar
  5. Chen, G.X., Asada, K.: Ascorbate peroxidases in tea leaves: occurrence of two isozymes and the differences in their enzymatic and molecular properties. — Plant Cell Physiol. 30: 987–998, 1989.Google Scholar
  6. Giannopolitis, C.N., Ries, S.K.: Superoxide dismutases. I. Occurrence in higher plants. — Plant Physiol. 59: 309–314, 1977.CrossRefPubMedGoogle Scholar
  7. Kavitha, K., Venkataraman, G., Parida, A.: An oxidative and salinity stress induced peroxisomal ascorbate peroxidase from Avicennia marina: molecular and functional characterization. — Plant Physiol. Biochem. 46: 794–804, 2008.CrossRefPubMedGoogle Scholar
  8. Kooten, O., Snel, J.F.H.: The use of chlorophyll fluorescence nomenclature in plant stress physiology. — Photosynth. Res. 25: 147–150, 1990.CrossRefGoogle Scholar
  9. Kratsch, H.A., Wise, R.R.: The ultrastructure of chilling stress. — Plant Cell Environ. 23: 337–350, 2000.CrossRefGoogle Scholar
  10. Lee, Y.P., Kim, S.H., Bang, J.W., Lee, H.S., Kwak, S.S., Kwon, S.Y.: Enhanced tolerance to oxidative stress in transgenic tobacco plants expressing three antioxidant enzymes in chloroplasts. — Plant Cell Rep. 26: 591–598, 2007.CrossRefPubMedGoogle Scholar
  11. Murgia, I., Tarantino, D., Vannini, C., Bracale, M., Carravieri, S., Soave, C.: Arabidopsis thaliana plants over-expressing thylakoidal ascorbate peroxidase show increased resistance to paraquat-induced photo-oxidative stress and to nitric oxide-induced cell death. — Plant J. 38: 940–953, 2004.CrossRefPubMedGoogle Scholar
  12. Robinson, S.P., Downton, W.J.S., Millhouse, J.A.: Photosynthesis and ion content of leaves and isolated chloroplasts of salt-stressed spinach. — Plant Physiol. 73: 238–242, 1983.CrossRefPubMedGoogle Scholar
  13. Sairam, P.K., Srivastava, G.C.: Changes in antioxidant activity in sub-cellular fractions of tolerant and susceptible wheat genotypes in response to long term salt stress. — Plant Sci. 162: 897–904, 2002.CrossRefGoogle Scholar
  14. Shi, W.M., Muramoto, Y., Ueda, A., Takabe, T.: Cloning of peroximal ascorbate peroxidase gene from barley and enhanced thermo-tolerance by over-expressing in Arabidopsis thaliana. — Gene 273: 23–27, 2001.CrossRefPubMedGoogle Scholar
  15. Shigeoka, S., Ishikawa, T., Tamoi, M., Miyagawa, Y., Takeda, T., Yabuta, Y., Yoshimura, K.: Regulation and function of ascorbate peroxidase isoenzymes. — J. exp. Bot. 53: 1305–1309, 2002.CrossRefPubMedGoogle Scholar
  16. Song, X.S., Hu, W.H., Mao, W.H., Ogweno, J.O., Zhou, Y.H., Yu, J.Q.: Response of ascorbate peroxidase isoenzymes and ascorbate regeneration system to abiotic stresses in Cucumis sativus L. — Plant Physiol. Biochem. 43: 1082–1088, 2005.CrossRefPubMedGoogle Scholar
  17. Wang, W.Q., Li, B., Meng, Q.W., Zou, Q.: The sequence of Lycopersicon esculentum thylakoid-bound ascorbate peroxidase gene TtAPX. — Plant Physiol. mol. Biol. 28: 491–492, 2002.Google Scholar
  18. Wang, Y., Yang, Z.M., Zhang, Q.F., Li, J.L.: Enhanced chilling tolerance in Zoysia matrella by pre-treatment with salicylic acid, calcium chloride, hydrogen peroxide or 6-benzylaminopurine. — Biol. Plant. 53: 179–182, 2009.CrossRefGoogle Scholar
  19. Yabuta, Y., Motoki, T., Yoshimura, K., Takeda, T., Ishikawa, T., Shigeoka, S.: Thylakoid membrane-bound peroxidase is a limiting factor of anti-oxidative systems under photo-oxidative stress. — Plant J. 32: 915–925, 2002.CrossRefPubMedGoogle Scholar
  20. Yoshimura, K., Yabuta, Y., Ishikawa, T., Shigeoka, S.: Expression of spinach ascorbate peroxidase isoenzymes in response to oxidative stresses. — Plant Physiol. 123: 223–233, 2000.CrossRefPubMedGoogle Scholar
  21. Zhou, Y.H., Yu, J.Q., Huang, L.F., Nogues, S.: The relationship between CO2 assimilation, photosynthetic electron transport and water-water cycle in chilling-exposed cucumber leaves under low light and subsequent recovery. — Plant Cell Environ. 27: 1503–1514, 2004.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • W. -H. Sun
    • 1
  • M. Duan
    • 1
  • F. Li
    • 1
  • D. -F. Shu
    • 1
  • S. Yang
    • 1
  • Q. -W. Meng
    • 1
  1. 1.College of Life Science, State Key Laboratory of Crop BiologyShandong Agricultural UniversityTai’anP.R. China

Personalised recommendations