Plant Cell Reports

, Volume 27, Issue 4, pp 767–778 | Cite as

Constitutive expression of a meiotic recombination protein gene homolog, OsTOP6A1, from rice confers abiotic stress tolerance in transgenic Arabidopsis plants

  • Mukesh Jain
  • Akhilesh K. Tyagi
  • Jitendra P. KhuranaEmail author
Biotic and Abiotic Stress


Plant productivity is greatly influenced by various environmental stresses, such as high salinity and drought. Earlier, we reported the isolation of topoisomerase 6 homologs from rice and showed that over expression of OsTOP6A3 and OsTOP6B confers abiotic stress tolerance in transgenic Arabidopsis plants. In this study, we have assessed the function of nuclear-localized topoisomerase 6 subunit A homolog, OsTOP6A1, in transgenic Arabidopsis plants. The over expression of OsTOP6A1 in transgenic Arabidopsis plants driven by cauliflower mosaic virus-35S promoter resulted in pleiotropic effects on plant growth and development. The transgenic Arabidopsis plants showed reduced sensitivity to stress hormone, abscisic acid (ABA), and tolerance to high salinity and dehydration at the seed germination; seedling and adult stages as reflected by the percentage of germination, fresh weight of seedlings and leaf senescence assay, respectively. Concomitantly, the expression of many stress-responsive genes was enhanced under various stress conditions in transgenic Arabidopsis plants. Moreover, microarray analysis revealed that the expression of a large number of genes involved in various processes of plant growth and development and stress responses was altered in transgenic plants. Although AtSPO11-1, the homolog of OsTOP6A1 in Arabidopsis, has been implicated in meiotic recombination; the present study demonstrates possible additional role of OsTOP6A1 and provides an effective tool for engineering crop plants for tolerance to different environmental stresses.


Abiotic stress Gene expression Rice (OryzasativaTopoisomerase 6 subunit A1 Transgenic Arabidopsis 



This work was supported financially by the Department of Biotechnology, Government of India, and the University Grants Commission, New Delhi. The award of Senior Research Fellowship to MJ from the Council of Scientific and Industrial Research is gratefully acknowledged.

Supplementary material

299_2007_491_MOESM1_ESM.tif (113 kb)
Fig. S1 (TIF 113 kb)
299_2007_491_MOESM2_ESM.doc (52 kb)
Table S1 (DOC 52 kb)
299_2007_491_MOESM3_ESM.doc (296 kb)
Table S2 (DOC 296 kb)
299_2007_491_MOESM4_ESM.doc (190 kb)
Table S3 (DOC 189 kb)
299_2007_491_MOESM5_ESM.doc (412 kb)
Table S4 (DOC 412 kb)


  1. Abe H, Yamaguchi-Shinozaki K, Urao T, Iwasaki T, Hosokawa D, Shinozaki K (1997) Role of Arabidopsis MYC and MYB homologs in drought- and abscisic acid-regulated gene expression. Plant Cell 9:1859–1868PubMedCrossRefGoogle Scholar
  2. Bergerat A, de Massy B, Gadelle D, Varoutas PC, Nicolas A, Forterre P (1997) An atypical topoisomerase II from Archaea with implications for meiotic recombination. Nature 386:414–417PubMedCrossRefGoogle Scholar
  3. Briolat V, Reysset G (2002) Identification of the Clostridium perfringens genes involved in the adaptive response to oxidative stress. J Bacteriol 184:2333–2343PubMedCrossRefGoogle Scholar
  4. Buhler C, Gadelle D, Forterre P, Wang JC, Bergerat A (1998) Reconstitution of DNA topoisomerase VI of the thermophilic archaeon Sulfolobus shibatae from subunits separately overexpressed in Escherichia coli. Nucleic Acids Res 26:5157–5162PubMedCrossRefGoogle Scholar
  5. Buhler C, Lebbink JH, Bocs C, Ladenstein R, Forterre P (2001) DNA topoisomerase VI generates ATP-dependent double-strand breaks with two-nucleotide overhangs. J Biol Chem 276:37215–37222PubMedCrossRefGoogle Scholar
  6. Celerin M, Merino ST, Stone JE, Menzie AM, Zolan ME (2000) Multiple roles of Spo11 in meiotic chromosome behavior. EMBO J 19:2739–2750PubMedCrossRefGoogle Scholar
  7. Champoux JJ (2001) DNA topoisomerases: structure, function, and mechanism. Annu Rev Biochem 70:369–413PubMedCrossRefGoogle Scholar
  8. Chen W, Provart NJ, Glazebrook J, Katagiri F, Chang HS, Eulgem T, Mauch F, Luan S, Zou G, Whitham SA, Budworth PR, Tao Y, Xie Z, Chen X, Lam S, Kreps JA, Harper JF, Si-Ammour A, Mauch-Mani B, Heinlein M, Kobayashi K, Hohn T, Dangl JL, Wang X, Zhu T (2002) Expression profile matrix of Arabidopsis transcription factor genes suggests their putative functions in response to environmental stresses. Plant Cell 14:559–574PubMedCrossRefGoogle Scholar
  9. Chu Z, Yuan M, Yao J, Ge X, Yuan B, Xu C, Li X, Fu B, Li Z, Bennetzen JL, Zhang Q, Wang S (2006) Promoter mutaions of an essential gene for pollen development result in disease resistance in rice. Genes Dev 20:1250–1255PubMedCrossRefGoogle Scholar
  10. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743PubMedCrossRefGoogle Scholar
  11. Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Report 1:19–21CrossRefGoogle Scholar
  12. Dernburg AF, McDonald K, Moulder G, Barstead R, Dresser M, Villeneuve AM (1998) Meiotic recombination in C. elegans initiates by a conserved mechanism and is dispensable for homologous chromosome synapsis. Cell 94:387–398PubMedCrossRefGoogle Scholar
  13. Gong Z, Lee H, Xiong L, Jagendorf A, Stevenson B, Zhu JK (2002) RNA helicase-like protein as an early regulator of transcription factors for plant chilling and freezing tolerance. Proc Natl Acad Sci USA 99:11507–11512PubMedCrossRefGoogle Scholar
  14. Grelon M, Vezon D, Gendrot G, Pelletier G (2001) AtSPO11–1 is necessary for efficient meiotic recombination in plants. EMBO J 20:589–600PubMedCrossRefGoogle Scholar
  15. Hartung F, Puchta H (2000) Molecular characterization of two paralogous SPO11 homologs in Arabidopsis thaliana. Nucleic Acids Res 28:1548–1554PubMedCrossRefGoogle Scholar
  16. Hartung F, Puchta H (2001) Molecular characterization of homologs of both subunits A (SPO11) and B of the archaebacterial topoisomerase 6 in plants. Gene 271:81–86PubMedCrossRefGoogle Scholar
  17. Hartung F, Angelis KJ, Meister A, Schubert I, Melzer M, Puchta H (2002) An archaebacterial topoisomerase homolog not present in other eukaryotes is indispensable for cell proliferation of plants. Curr Biol 12:1787–1791PubMedCrossRefGoogle Scholar
  18. Hettiarachchi GH, Reddy MK, Sopory SK, Chattopadhyay S (2005) Regulation of TOP2 by various abiotic stresses including cold and salinity in pea and transgenic tobacco plants. Plant Cell Physiol 46:1154–1160PubMedCrossRefGoogle Scholar
  19. Jain M, Tyagi SB, Thakur JK, Tyagi AK, Khurana JP (2004) Molecular characterization of a light-responsive gene, breast basic conserved protein 1 (OsiBBC1), encoding nuclear-localized protein homologous to ribosomal protein L13 from Oryza sativa indica. Biochim Biophys Acta 1676:182–192PubMedGoogle Scholar
  20. Jain M, Kaur N, Tyagi AK, Khurana JP (2006a) The auxin-responsive GH3 gene family in rice (Oryza sativa). Funct Integr Genomics 6:36–46PubMedCrossRefGoogle Scholar
  21. Jain M, Tyagi AK, Khurana JP (2006b) Overexpression of putative topoisomearse 6 genes from rice confers stress tolerance in transgenic Arabidopsis plants. FEBS J 273:5245–5260PubMedCrossRefGoogle Scholar
  22. Jain M, Nijhawan A, Arora R, Agarwal P, Ray S, Sharma P, Kapoor S, Tyagi AK, Khurana JP (2007) F-box proteins in rice. Genome-wide analysis, classification, temporal and spatial gene expression during panicle and seed development, and regulation by light and abiotic stress. Plant Physiol 143:1467–1483PubMedCrossRefGoogle Scholar
  23. Jonak C, Kiegerl S, Ligterink W, Barker PJ, Huskisson NS, Hirt H (1996) Stress signaling in plants: a mitogen-activated protein kinase pathway is activated by cold and drought. Proc Natl Acad Sci USA 93:11274–11279PubMedCrossRefGoogle Scholar
  24. Kasuga M, Liu Q, Miura S, Yamaguchi-Shinozaki K, Shinozaki K (1999) Improving plant drought, salt, and freezing tolerance by gene transfer of a single stress-inducible transcription factor. Nat Biotechnol 17:287–291PubMedCrossRefGoogle Scholar
  25. Kasuga M, Miura S, Shinozaki K, Yamaguchi-Shinozaki K (2004) A combination of the Arabidopsis DREB1A gene and stress-inducible rd29A promoter improved drought- and low-temperature stress tolerance in tobacco by gene transfer. Plant Cell Physiol 45:346–350PubMedCrossRefGoogle Scholar
  26. Keeney S, Giroux CN, Kleckner N (1997) Meiosis-specific DNA double-strand breaks are catalyzed by Spo11, a member of a widely conserved protein family. Cell 88:375–384PubMedCrossRefGoogle Scholar
  27. Kiyosue T, Yamaguchi-Shinozaki K, Shinozaki K (1994) Characterization of two cDNAs (ERD10 and ERD14) corresponding to genes that respond rapidly to dehydration stress in Arabidopsis thaliana. Plant Cell Physiol 35:225–231PubMedGoogle Scholar
  28. Kovtun Y, Chiu WL, Tena G, Sheen J (2000) Functional analysis of oxidative stress-activated mitogen-activated protein kinase cascade in plants. Proc Natl Acad Sci USA 97:2940–2945PubMedCrossRefGoogle Scholar
  29. Lan L, Li M, Lai Y, Xu W, Kong Z, Ying K, Han B, Xue Y (2005) Microarray analysis reveals similarities and variations in genetic programs controlling pollination/fertilization and stress responses in rice (Oryza sativa L.). Plant Mol Biol 59:151–164PubMedCrossRefGoogle Scholar
  30. Laxmi A, Paul LK, Peters JL, Khurana JP (2004) Arabidopsis constitutive photomorphogenic mutant, bls1, displays altered brassinosteroid response and sugar sensitivity. Plant Mol Biol 56:185–201PubMedCrossRefGoogle Scholar
  31. Liu HY, Nefsky BS, Walworth NC (2002) The Ded1 DEAD box helicase interacts with Chk1 and Cdc2. J Biol Chem 277:2637–2643PubMedCrossRefGoogle Scholar
  32. McKim KS, Hayashi-Hagihara A (1998) mei-W68 in Drosophila melanogaster encodes a Spo11 homolog: evidence that the mechanism for initiating meiotic recombination is conserved. Genes Dev 12:2932–2942PubMedCrossRefGoogle Scholar
  33. Mukhopadhyay A, Vij S, Tyagi AK (2004) Overexpression of a zinc-finger protein gene from rice confers tolerance to cold, dehydration, and salt stress in transgenic tobacco. Proc Natl Acad Sci USA 101:6309–6314PubMedCrossRefGoogle Scholar
  34. Nichols MD, DeAngelis K, Keck JL, Berger JM (1999) Structure and function of an archaeal topoisomerase VI subunit with homology to the meiotic recombination factor Spo11. EMBO J 18:6177–6188PubMedCrossRefGoogle Scholar
  35. Potocky M, Elias M, Profotova B, Novotna Z, Valentova O, Zarsky V (2003) Phosphatidic acid produced by phospholipase D is required for tobacco pollen tube growth. Planta 217:122–130PubMedGoogle Scholar
  36. Saijo Y, Hata S, Kyozuka J, Shimamoto K, Izui K (2000) Over-expression of a single Ca2+-dependent protein kinase confers both cold and salt/drought tolerance on rice plants. Plant J 23:319–327PubMedCrossRefGoogle Scholar
  37. Sanan-Mishra N, Pham XH, Sopory SK, Tuteja N (2005) Pea DNA helicase 45 overexpression in tobacco confers high salinity tolerance without affecting yield. Proc Natl Acad Sci USA 102:509–514PubMedCrossRefGoogle Scholar
  38. Shinozaki K, Yamaguchi-Shinozaki K (1997) Gene expression and signal transduction in water-stress response. Plant Physiol 115:327–334PubMedCrossRefGoogle Scholar
  39. Stacey NJ, Kuromori T, Azumi Y, Roberts G, Breuer C, Wada T, Maxwell A, Roberts K, Sugimoto-Shirasu K (2006) Arabidopsis SPO11–2 functions with SPO11-1 in meiotic recombination. Plant J 48:206–216PubMedCrossRefGoogle Scholar
  40. Sugimoto-Shirasu K, Stacey NJ, Corsar J, Roberts K, McCann MC (2002) DNA topoisomerase VI is essential for endoreduplication in Arabidopsis. Curr Biol 12:1782–1786PubMedCrossRefGoogle Scholar
  41. Veena, Reddy VS, Sopory SK (1999) Glyoxalase I from Brassica juncea: molecular cloning, regulation and its over-expression confer tolerance in transgenic tobacco under stress. Plant J 17:385–395PubMedCrossRefGoogle Scholar
  42. Wang JC (2002a) Cellular roles of DNA topoisomerases: a molecular perspective. Nat Rev Mol Cell Biol 3:430–440PubMedCrossRefGoogle Scholar
  43. Wang X (2002b) Phospholipase D in hormonal and stress signaling. Curr Opin Plant Biol 5:408–414PubMedCrossRefGoogle Scholar
  44. Westermarck J, Weiss C, Saffrich R, Kast J, Musti AM, Wessely M, Ansorge W, Seraphin B, Wilm M, Valdez BC, Bohmann D (2002) The DEXD/H-box RNA helicase RHII/Gu is a co-factor for c-Jun-activated transcription. EMBO J 21:451–460PubMedCrossRefGoogle Scholar
  45. Xu D, Duan X, Wang B, Hong B, Ho T, Wu R (1996) Expression of a late embryogenesis abundant protein gene, HVA1, from barley confers tolerance to water deficit and salt stress in transgenic rice. Plant Physiol 110:249–257PubMedGoogle Scholar
  46. Yamaguchi-Shinozaki K, Shinozaki K (1993) Arabidopsis DNA encoding two desiccation-responsive rd29 genes. Plant Physiol 101:1119–1120PubMedCrossRefGoogle Scholar
  47. Yin Y, Cheong H, Friedrichsen D, Zhao Y, Hu J, Mora-Garcia S, Chory J (2002) A crucial role for the putative Arabidopsis topoisomerase VI in plant growth and development. Proc Natl Acad Sci USA 99:10191–10196PubMedCrossRefGoogle Scholar
  48. Zhu J, Shi H, Lee BH, Damsz B, Cheng S, Stirm V, Zhu JK, Hasegawa PM, Bressan RA (2004) An Arabidopsis homeodomain transcription factor gene, HOS9, mediates cold tolerance through a CBF-independent pathway. Proc Natl Acad Sci USA 101:9873–9878PubMedCrossRefGoogle Scholar
  49. Zhu J, Verslues PE, Zheng X, Lee BH, Zhan X, Manabe Y, Sokolchik I, Zhu Y, Dong CH, Zhu JK, Hasegawa PM, Bressan RA (2005) HOS10 encodes an R2R3-type MYB transcription factor essential for cold acclimation in plants. Proc Natl Acad Sci USA 102:9966–9971PubMedCrossRefGoogle Scholar
  50. Zonia L, Munnik T (2004) Osmotically induced cell swelling versus cell shrinking elicits specific changes in phospholipid signals in tobacco pollen tubes. Plant Physiol 134:813–823PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Mukesh Jain
    • 1
  • Akhilesh K. Tyagi
    • 1
  • Jitendra P. Khurana
    • 1
    Email author
  1. 1.Interdisciplinary Centre for Plant Genomics and Department of Plant Molecular BiologyUniversity of Delhi South CampusNew DelhiIndia

Personalised recommendations