Skip to main content
Log in

Changes in expression of the cytosolic ascorbate peroxidase gene,ca-capx1, during germination and development of hot pepper seedlings

  • Published:
Journal of Plant Biology Aims and scope Submit manuscript

Abstract

Ascorbate peroxidase (APX), an antioxidant enzyme, scavenges H2O2 that is produced by normal metabolism and cellular oxidative stresses. To investigate its role during germination and seedling growth, we isolated a cDNA encoding cytosolic APX (cAPX) in hot pepper(Capsicum annuum L). The full-length clone,Ca- cAPX1, is 1011 bp long and has an ORF encoding 249 amino acid residues. During seedling development, cAPX activity and expression levels were higher at Days 5 and 6 post-imbibition, respectively, whereas those of catalase (CAT) increased at Days 8 and 10. The increased amount of H2O2 in that early developmental stage (Day 5) may have been counteracted mainly by APX, and further removed by CAT in cooperation with APX. To determine whether the accumulation of H2O2 via suppression ofcAPX expression might be a factor in stimulating germination, we constructed a transformant ofCaAPX1. Compared with the wild type, the germination rate for the antisense-suppressedArabidopsis increased by 26%, while its H2O2 content rose by 50%. Therefore, we propose that the pre-germination suppression ofcAPX expression stimulates seed germination by promoting the accumulation of H2O2.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Literature cited

  • Asada K (1993) Divergence of peroxide-scavenging peroxidases in organisms,In K Yaki, ed, Active Oxygens, Lipid Peroxides and Antioxidants. CRC Press, Boca Raton, pp 289–298

    Google Scholar 

  • Bewley JD, Black M (1994) Seeds. Physiology of Development and Germination, Ed 2. Plenum Press, New York, pp 248–252

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem72: 248–254

    Article  PubMed  CAS  Google Scholar 

  • Chance B, Sies H, Boveris A (1979) Hydroperoxide metabolism in mammalian organs. Physiol Rev59: 527–605

    PubMed  CAS  Google Scholar 

  • Chien CT, Lin TP (1994) Mechanism of hydrogen peroxide in improving the germination ofCinnamonum camphora seed. Seed Sci Technol22: 231–236

    Google Scholar 

  • Clough SJ, Bent AF (1998) Floral dip: A simplified method forAgrobacterium mediated transformation ofArabidopsis thaliana. Plant J16: 735–743

    Article  PubMed  CAS  Google Scholar 

  • de Gara L, de Pinto MC, Arrigoni O (1997) Ascorbate synthesis and ascorbate peroxidase activity during the early stage of wheat germination. Physiol Plant100: 894–900

    Article  Google Scholar 

  • de Tullio M, Arrigoni O (1992) Ascorbate peroxidase activity in resistant and susceptible plants ofLycopersicon esculentum. Boll Soc Ital Biol Sper68: 613–617

    PubMed  Google Scholar 

  • Everse J, Everse KE, Grisham MB (1991) Peroxidases in Chemistry and Biology, Vol 1. CRC Press, Boca Raton

    Google Scholar 

  • Fath A, Bethke PC, Jones RL (2001) Enzymes that scavenge reactive oxygen species are down-regulated prior to gibberellic acid-induced programmed cell death in barley aleurone. Plant Physiol126: 156–166

    Article  PubMed  CAS  Google Scholar 

  • Fontaine O, Huault C, Pavis N, Billard JP (1994) Dormancy breakage ofHordeum vulgare seeds: Effects of hydrogen peroxide and scarification on glutathione level and glutathione reductase activity. Plant Physiol Biochem32: 677–683

    CAS  Google Scholar 

  • Hendry GAF (1993) Oxygen, free radical processes and seed longevity. Seed Sci Res3: 141–153

    Article  CAS  Google Scholar 

  • Jimenez A, Hernandez JA, del Río LA, Sevilla F (1997) Ascorbate-glutathione cycle in mitochondria and peroxisomes of pea leaves: Changes induced by leaf senescence. Phyton37: 101–108

    CAS  Google Scholar 

  • Kim JH, Baek MH, Chung BY, Wi SG, Kim JS (2004) Alterations in the photosynthetic pigments and antioxidant machineries of red pepper(Capsicum annuum L.) seedlings from gamma-irradiated seeds. J Plant Biol47: 314–321

    Article  CAS  Google Scholar 

  • Kim YG, Kwon SI, An CS (1977) Isolation and characterization of cytosolic copper/zinc Superoxide dismutase fromCapsicum annuum L. Mol Cells7: 668–673

    Google Scholar 

  • Klapheck S, Zimmer I, Cosse H (1990) Scavenging of hydrogen peroxide in the endosperm ofRicinus communis by ascorbate peroxidase. Plant Cell Physiol31: 1005–1013

    CAS  Google Scholar 

  • Kwon SI, An CS (2001) Molecular cloning, characterization and expression analysis of a catalase cDNA from hot pepper(Capsicum annuum L). Plant Sci160: 961–969

    Article  PubMed  CAS  Google Scholar 

  • Mittler R, Zilinskas BA (1991) Purification and characterization of pea cytosolic ascorbate peroxidase. Plant Physiol97: 962–968

    Article  PubMed  CAS  Google Scholar 

  • Mittler R, Zilinskas BA (1993) Detection of ascorbate peroxidase activity in native gels by inhibition of the ascorbate-dependent reduction of nitroblue tetrazolium. Anal Biochem212: 540–546

    Article  PubMed  CAS  Google Scholar 

  • Naredo MEB, Juliano AB, Lu BR, de Guzman F, Jackson MT (1998) Response to seed dormancy-breaking treatments in rice species(Oryza sativa L.). Seed Sci Technol26: 675–689

    Google Scholar 

  • Ogawa K, Iwabuchi M (2001) A mechanism for promoting the germination ofZinnia elegans seeds by hydrogen peroxide. Plant Cell Physiol42: 286–291

    Article  PubMed  CAS  Google Scholar 

  • Patterson WR, Poulos TL (1995) Characterization and crystallization of recombinant pea cytosolic ascorbate peroxidase. J Biol Chem269: 17020–17024

    Google Scholar 

  • Puntarulo S, Galleano M, Sanchez RA, Boveris A (1991) Superoxide anion and hydrogen peroxide metabolism in soybean embryonic axes during germination. Biochim Biophysv Acta1074: 277–283

    CAS  Google Scholar 

  • Puntarulo S, Sanchez RA, Boveris A (1988) Hydrogen peroxide metabolism in soybean embryonic axes at the onset of germination. Plant Physiol86: 626–630

    Article  PubMed  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, NY

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA74: 5463–5467

    Article  PubMed  CAS  Google Scholar 

  • Schopfer P, Plachy C, Frahry G (2001) Release of reactive oxygen intermediates (Superoxide radicals, hydrogen peroxide, and hydroxyl radicals) and peroxidase in germinating radish seeds controlled by light, gibberellin, and abscisic acid. Plant Physiol125: 1591–1602

    Article  PubMed  CAS  Google Scholar 

  • Tommasi F, Paciolla C, D’Elia F, Arrigoni O (1995) Changes of ascorbate peroxidase and dehydroascorbate reductase activities during seed development ofVicia faba L. Boll Soc Ital Biol Sper71: 11–12

    Google Scholar 

  • Tommasi F, Paciolla C, de Pinto MC, de Gara L (2001) A comparative study of glutathione and ascorbate metabolism during germination ofPinus pinea L. seeds. J Exp Bot52: 1647–1654

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Chung Sun An.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Moon, S.C., Kvvon, S.I. & An, C.S. Changes in expression of the cytosolic ascorbate peroxidase gene,ca-capx1, during germination and development of hot pepper seedlings. J. Plant Biol. 48, 276–283 (2005). https://doi.org/10.1007/BF03030523

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF03030523

Keywords

Navigation