Plant Molecular Biology

, Volume 36, Issue 6, pp 833–845 | Cite as

The expression of a peroxiredoxin antioxidant gene, AtPer1, in Arabidopsis thaliana is seed-specific and related to dormancy

  • Camilla Haslekås
  • Robin A. P. Stacy
  • Vigdis Nygaard
  • Francisco A. Culiáñez-Macià
  • Reidunn B. Aalen


We have isolated a gene, AtPer1, from the dicotyledon Arabidopsis thaliana, which shows similarity to the 1-cysteine (1-Cys) peroxiredoxin family of antioxidants. In higher plants, members of this group of antioxidants have previously only been isolated from monocotyledons. It has been suggested that seed peroxiredoxins protect tissues from reactive oxygen species during desiccation and early imbibition and/or are involved in the maintenance of/protection during dormancy. AtPer1 expression is restricted to seeds. Despite differences in seed development between monocots and dicots, AtPer1 shows an expression pattern during seed development and germination similar to the dormancy-related transcript Per1 in barley. In situ hybridization identifies AtPer1 as the first aleurone-expressed transcript characterized in developing Arabidopsis seeds. The transcript is also expressed in the embryo. AtPer1 expression in seeds is unaltered in an ABA-deficient mutant (aba-1) during seed development, while expression in seeds of an ABA-insensitive mutant (abi3-1) is reduced. The transcript is not induced in vegetative tissue in response to stress by ABA or drought. AtPer1 transcript levels are correlated to germination frequencies of wildtype seeds, but AtPer1 transcript abundance is not sufficient for expression of dormancy in non-dormant mutants. Hypotheses on peroxiredoxin function are discussed in view of the results presented here.

embryo- and aleurone-specific expression desiccation tolerance thiol-specific antioxidant (TSA) late embryogenesis-abundant (lea) aba-1 and abi3-1 mutants 


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  1. 1.
    Aalen RB, Opsahl-Fernstad HG, Linnestad C, Olsen OA: Transcripts encoding an oleosin and a dormancyrelated protein are present in both the aleurone layer and the embryo of developing barley (Hordeum vulgare L.) seeds. Plant J 5: 385–396 (1994).PubMedGoogle Scholar
  2. 2.
    Baier M, Dietz KJ: Primary structure and expression of plant homologues of animal and fungal thioredoxindependent peroxide reductases and bacterial alkyl hydroperoxide reductases. Plant Mol Biol 31: 553–564 (1996).PubMedGoogle Scholar
  3. 3.
    Baker J, Steele C, Dure L III: Sequence and characterization of 6 Lea proteins and their genes from cotton. Plant Mol Biol 11: 277–291 (1988).Google Scholar
  4. 4.
    Bergfeld R, Schopfer P: Differentiation of a functional aleurone layer within the seed coat of Sinapis alba L. Ann Bot 57: 25–33 (1986).Google Scholar
  5. 5.
    Bowman JL, Mansfield SG: Embryogenesis: An Introduction. In: Bowman JL (ed) Arabidopsis: An Atlas of Morphology and Development, pp. 351–361. SpringerVerlag, New York (1994).Google Scholar
  6. 6.
    Cakmak I, Strbac D, Marschner H: Activities of hydrogen peroxidescavenging enzymes in germinating wheat seeds. J Exp Bot 258: 127–132 (1993).Google Scholar
  7. 7.
    Chae HZ, Robinson K, Pool LB, Church G, Storz G, Rhee SG: Cloning and sequencing of thiolspecific antioxidant from mammalian brain: alkyl hydroperoxide reductase and thiolspecific antioxidant define a large family of antioxidant enzymes. Proc Natl Acad Sci USA 91: 7017–7020 (1994).Google Scholar
  8. 8.
    Chae HZ, Uhm TB, Rhee SG: Dimerization of thiolspecific antioxidant and the essential role of cystein-47. Proc Natl Acad Sci USA 91: 7022–7026 (1994).Google Scholar
  9. 9.
    Chae HZ, Chung SJ, Rhee SG: Thioredoxindependent peroxide reductase from yeast. J Biol Chem 269: 27670–27678 (1994).PubMedGoogle Scholar
  10. 10.
    Chen ZL, Schuler MA, Beachy RN: Functional analysis of regulatory elements in a plant embryospecific gene. Proc Natl Acad Sci USA 83: 8560–8564 (1986).PubMedGoogle Scholar
  11. 11.
    Dellaporta SL, Wood J, Hicks JB: A plant DNA minipreparation: version II. Plant Mol Biol Rep 1: 19–21 (1983).Google Scholar
  12. 12.
    Dure III L, Crouch M, Harada J, Ho THD, Mundy J, Quatrano R, Thomas T, Sung ZR: Common amino acid sequence domains among the LEA proteins of higher plants. Plant Mol Biol 12: 475–486 (1989).Google Scholar
  13. 13.
    Espelund M, De Bedout J A, Outlaw W H, Jakobsen K S: Environmental and hormonal regulation of barley lateembryogenesisabundant (Lea) mRNAs is via different signal transduction pathways. Plant Cell Env 18: 943–949 (1995).Google Scholar
  14. 14.
    Espelund M, Stacy PRA, Jakobsen KS: A simple method for generating singlestranded DNA probes labeled to high activities. Nucl Acids Res 18: 6157–6158 (1990).PubMedGoogle Scholar
  15. 15.
    Feinbaum RL, Ausubel FM: Transcriptional regulation of the Arabidopsis thaliana chalcone synthase gene. Mol Cell Biol 8: 1985–1992 (1988).PubMedGoogle Scholar
  16. 16.
    Finkelstein RR: Abscisic acid–insensitive mutations provide evidence for stagespecific signal pathways regulating expression of anArabidopsis late embryogenesisabundant (lea) gene. Mol Gen Genet 238: 401–408 (1993).CrossRefPubMedGoogle Scholar
  17. 17.
    Finkelstein R, Somerville C: Three classes of abscisic acid (ABA)insensitive mutations of Arabidopsis define genes that control overlapping subsets of ABA responses. Plant Physiol 94: 1172–1179 (1990).Google Scholar
  18. 18.
    Frugoli JA, Zhong HH, Nuccio ML, McCourt P, McPeek MA, Thomas TL, Robertson McClung C: Catalase is encoded by a multigene family in Arabidopsis thaliana (L.) Heynh. Plant Physiol 112: 327–336 (1996).PubMedGoogle Scholar
  19. 19.
    Galau GA, Bijaisoradat N, Hughes DW: Accumulation kinetics of cotton late embryogenesisabundant mRNAs and storage protein mRNAs: coordinate regulation during embryogenesis and the role of abscisic acid. Devel Biol 123: 198–212 (1987).Google Scholar
  20. 20.
    Galau G, Hughes D, Dure L III: Abscicic acid induction of cloned cotton late embryogenesisabundant (Lea) mRNAs. Plant Mol Biol 7: 155–170 (1986).Google Scholar
  21. 21.
    Goday A, Jensen AB, Culiáñez-Macià FA, Albà MM, Figueras M, Serratosa J, Torrent M, Pagès M: The maize abscisic acidresponsive protein Rab17 is located in the nucleus and interacts with nuclear localization signals. Plant Cell 6: 351–360 (1994).CrossRefPubMedGoogle Scholar
  22. 22.
    Goldmark PJ, Curry J, Morris CF, WalkerSimmons MK: Cloning and expression of an embryospecific mRNA upregulated in hydrated dormant seeds. Plant Mol Biol 19: 433–441 (1992).CrossRefPubMedGoogle Scholar
  23. 23.
    Guan L, Scandalios JG: Characterization of the catalase antioxidant defense gene Cat1 of maize, and its developmentally regulated expression in transgenic tobacco. Plant J 3: 527–536 (1993).CrossRefPubMedGoogle Scholar
  24. 24.
    Guiltinan MJ, Marcotte WR Jr, Quatrano RS: A plant leucine zipper protein that recognizes an abscisic acid response element. Nature 250: 267–271 (1990).Google Scholar
  25. 25.
    Hole DJ, Smith JD, Cobb BG: Regulation of embryo dormancy by manipulation of abscisic acid in kernels and associated cob tissue of Zea mays L. cultured in vitro. Plant Physiol 91: 101–105 (1989).Google Scholar
  26. 26.
    HolstJensen A, Kohn LM, Jakobsen KS, Schumacher T: Molecular phylogeny and evolution of Monilinia (sclerotiniaceae) based on coding and non coding rDNA sequences. Am J Bot (in press).Google Scholar
  27. 27.
    Hughes DW, Galau GA: Temporally modular gene expression during cotyledon development. Genes Devel 3: 358–369 (1989).PubMedGoogle Scholar
  28. 28.
    Jacobson FS, Morgans RW, Christman MF, Ames BN: An alkyl hydroperoxide reductase from Salmonella typhimurium involved in the defence of DNA against oxidative damage. J Biol Chem 264: 1488–1496 (1989).PubMedGoogle Scholar
  29. 29.
    Jakobsen KS, Breivold E, Hornes E: Purification of mRNA directly from crude plant tissues in 15 minutes using magnetic oligo dT microspheres. Nucl Acids Res 18: 3669 (1990).PubMedGoogle Scholar
  30. 30.
    Jakobsen KS, Haugen M, Sæbøe-Larsen S, Hollung K, Espelund M, Hornes E: DirectmRNAisolation usingmagnetic oligo dT beads: A protocol for all types of cell cultures, animal and plant tissues. In: Uhlen M, Hornes E, Olsvik, Ø(eds) Advances in Biomagnetic Separation, pp. 61–71. Eaton Publishers, MA, USA (1994).Google Scholar
  31. 31.
    Jones PL, Kucera G, Gordon H, Boss JM: Cloning and characterization of the murine manganous superoxide dismutaseencoding gene. Gene 153: 155–161 (1995).PubMedGoogle Scholar
  32. 32.
    Karssen CM, Brinkhorstvan der Swan DLC, Breekland AE, Koornneef M: Induction of dormancy during seed development by endogenous abscisic acid: studies on abscisic acid deficient genotypes of Arabidopsis thaliana (L.) Heynh. Planta 157: 158–165 (1983).Google Scholar
  33. 33.
    Kawagoe Y, Campell BR, Murai N: Synergism between the CACGTG (G-box) and the CACCTG ciselements is required for activation of the bean seed storage protein β-phasoline gene. Plant J 5: 885–890 (1994).CrossRefPubMedGoogle Scholar
  34. 34.
    Kim K, Kim IH, Lee KY, Rhee SG, Stadtman E: The isolation and purification of a specific ‘protector’ protein which inhibits enzyme inactivation by a thiol/Fe(III)/O2 mixedfunction oxidation system. J Biol Chem 263: 4704–4711 (1988).PubMedGoogle Scholar
  35. 35.
    Koornneef M, Jorna ML, Brinkhorstvan Swan DLC, Karssen CM: The isolation of abscisic acid (ABA) deficient mutants by selection of induced revertants in nongerminating gibberellin sensitive lines of Arabidopsis thaliana (L.) Heynh. Theor Appl Genet 61: 385–393 (1982).Google Scholar
  36. 36.
    Koornneef M, Reuling G, Karssen CM: The isolation and characterization of abscisic acidinsensitive mutants of Arabidopsis thaliana. Physiol Plant 61: 377–383 (1984).Google Scholar
  37. 37.
    Leah R, Skriver K, Knudsen S, RuudHansen J, Raikhel NV, Mundy J: Identification of an enhancer/silencer sequence directing the aleuronespecific expression of a barley chitinase gene. Plant J 6: 579–589 (1994).PubMedGoogle Scholar
  38. 38.
    Leprince O, Hendry GAF, McKersie BD: The mechanisms of desiccation tolerance in developing seeds. Seed Sci Res 3: 231–246 (1993).Google Scholar
  39. 39.
    Lim YS, Cha MK, Yun CH, Kim HK, Kim K, Kim IH: Purification and characterization of thiolspecific antioxidant protein from human red blood cell: a new type of antioxidant protein. Biochem Biophys Res Comm 199: 199–206 (1994).CrossRefPubMedGoogle Scholar
  40. 40.
    Lim YS, Cha MK, Kim HK, Uhm TB, Park JW, Kim K, Kim IH: Removals of hydrogen peroxide and hydroxyl radical by thiolspecific antioxidant protein as a possible role in vivo. Biochem Biophys Res Comm 192: 273–280 (1993).CrossRefPubMedGoogle Scholar
  41. 41.
    Lindsey K and Topping JF: Embryogenesis: a question of pattern. J Exp Bot 259: 359–374 (1993).Google Scholar
  42. 42.
    Lång V, Palva TE: The expression of a rabrelated gene, Rab18, is induced by abscisic acid during the cold acclimation process of Arabidopsis thaliana (L.) Heynh. Plant Mol Biol 20: 951–962 (1992).PubMedGoogle Scholar
  43. 43.
    Melan MA, Enriquez ALD, Peterman TK: The LOX1 Gene of Arabidopsis is temporally and spatially regulated in germinating seedlings. Plant Physiol 105: 385–393 (1994).PubMedGoogle Scholar
  44. 44.
    Merry J: Studies on the embryo of Hordeum sativumI. The development of the embryo. Bull Torrey Club 68: 585–598 (1941).Google Scholar
  45. 45.
    Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497 (1962).Google Scholar
  46. 46.
    Netto L, Chae HZ, Kang SW, Rhee SG, Stadtman ER: Removal of hydrogen peroxide by thiolspecific antioxidant enzymes (TSA) is involved with its antioxidant propertiesTSA possesses thiol peroxidase activity. J Biol Chem 271: 15315–15321 (1996).PubMedGoogle Scholar
  47. 47.
    Pan SM, Yau YY: Characterization of superoxide dismutase in Arabidopsis. Taiwania 37: 58–66 (1992).Google Scholar
  48. 48.
    Parcy F, Valon C, Raynal M, GaubierComelia P, Delseny M, Giraudat J: Regulation of gene expression programs during Arabidopsis seed development: roles of the ABI3 locus and of endogenous abscisic acid. Plant Cell 6: 1567–1582 (1994).CrossRefPubMedGoogle Scholar
  49. 49.
    Puntarulo S, Galleano M, Sanchez RA, Boveris A: Superoxide anion and hydrogen peroxide metabolism in soybean embryonic axes during germination. Biochim Biophys Acta 1074: 277–283 (1991).PubMedGoogle Scholar
  50. 50.
    Rao MV, Gopinadhan P, Ormrod DP: UltravioletBand ozoneinduced biochemical changes in antioxidant enzymes of Arabidopsis thaliana. Plant Physiol 110: 125–136 (1996).CrossRefPubMedGoogle Scholar
  51. 51.
    Rushmore TH, Morton MR, Pickett CB: The antioxidant responsive element. J Biol Chem 266: 11632–11639 (1991).PubMedGoogle Scholar
  52. 52.
    Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory manual, 2nd ed. Cold Harbor Laboratory Press, Cold Spring, Harbor, NY (1989).Google Scholar
  53. 53.
    Scandalios JG: The antioxidant enzyme genes Cat and Sod of maize: regulation, functional significance, and molecular biology. In: Rattazzi MC, Scandalios JG, Whitt GS (eds) Isozymes: Current Topics in Biological and Medical Research, vol. 14: Molecular and Cellular Biology, pp. 19–44. Alan R. Liss, New York. (1987).Google Scholar
  54. 54.
    Scandalios JG: Response of plant antioxidant defense genes to environmental stress. Adv Genet 28: 1–41 (1990).Google Scholar
  55. 55.
    Scott II HB, Oliver MJ: Accumulation and polysomal recruitment of transcripts in response to desiccation and rehydration of the moss Tortula ruralis. J Exp Bot 45: 577–583 (1994).Google Scholar
  56. 56.
    Shirley BW, Hanley S, Goodman HM: Effects of ionizing radiation on a plant genome: Analysis of two Arabidopsis transparent testa mutations. Plant Cell 4: 333–347 (1992).CrossRefPubMedGoogle Scholar
  57. 57.
    Skriver K, Mundy J: Gene expression in response to abscisic acid and osmotic stress. Plant Cell 2: 503–512 (1990).CrossRefPubMedGoogle Scholar
  58. 58.
    Stacy JE, Ims RA, Stenseth NC, Jakobsen KS: Fingerprinting of diverse species with DNA probes generated from immobilized singlestranded DNA templates. Nucl Acids Res 19: 4404 (1991).Google Scholar
  59. 59.
    Stacy RAP, Munthe E, Steinum T, Sharma B, Aalen RB: A peroxiredoxin antioxidant is encoded by a dormancyrelated gene, Per1, expressed during late development in the aleurone and embryo of barley grains. Plant Mol Biol 31: 1205–1216 (1996).PubMedGoogle Scholar
  60. 60.
    Vaughan JG, Whitehouse JM: Seed structure and the taxonomy of the cruciferae. Bot J Linn Soc 64: 383–409 (1971).Google Scholar
  61. 61.
    WalkerSimmons M: ABA levels and sensitivity in developing wheat embryos of sprouting resistante and susceptible cultivars. Plant Physiol 84: 61–66 (1987).Google Scholar
  62. 62.
    Wang M, HeimovaaraDijkstra S, van Duijn B: Modulation of germination of embryos isolated from dormant and nondormant grains by manipulation of endogenous abscisic acid. Planta 195: 586–592 (1995).Google Scholar
  63. 63.
    Zong HH, Young JC, Pease EA, Hangarter RP, Robertson MC: Interactions between light and the circadian clock in the regulation of CAT2 expression in Arabidopsis. Plant Physiol 104: 889–898 (1994).PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • Camilla Haslekås
    • 1
  • Robin A. P. Stacy
    • 1
  • Vigdis Nygaard
    • 1
  • Francisco A. Culiáñez-Macià
    • 2
  • Reidunn B. Aalen
    • 1
  1. 1.Division of General GeneticsUniversity of OsloBlindern, 0315Norway
  2. 2.Instituto de Biologia Molecular y Celular de PlantasUniversidad Politécnica de Valencia, Camino de Vera s/nValenciaSpain

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