Skip to main content

Advertisement

Log in

Isolation of a novel RNA-dependent RNA polymerase 6 from Nicotiana glutinosa, NgRDR6, and analysis of its response to biotic and abiotic stresses

  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

RNA-dependent RNA polymerases (RDRs) play an important role in RNA silencing, antiviral and developmental progress. Here, we firstly isolated the full-length cDNA, genomic DNA and 5′-flanking region of RDR6 from Nicotiana glutinosa (NgRDR6). Sequences analysis revealed that the cDNA of NgRDR6 was 3,921 bp in length, and the deduced protein consisted of 1,197 amino acids, containing all highly conserved sequence motifs that are present among all RDRs families. Moreover, two introns were detected in the genomic sequences. We also firstly investigated the expression profiles of plant RDR6 under the treatments of gibberellin A (GA), H2O2, methyl jasmonate (MeJA), Potato virus Y (PVY), Tobacco mosaic virus (TMV), Cucumber mosaic virus (CMV), Rhizoctonia Solani and Colletotrichum nicotianae. In addition, the expression patterns of RDR6 in Nicotiana glutinosa under the treatments of salicylic acid (SA) and abscisic acid (ABA) were also been analyzed. The results indicated that the NgRDR6 mRNA accumulation could be induced by ABA, GA, MeJA, CMV, Rhizoctonia Solani and Colletotrichum nicotianae. In contrast, the expression level of NgRDR6 exhibited no remarkable difference under the treatments of PVY, TMV, H2O2 and SA. Further investigation suggested several potential cis-acting elements were found in the 5′-flanking sequence of NgRDR6, which might be responsible for the enhanced response to phytohormones.

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.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Hannon GJ (2002) RNA interference. Nature 418:244–251

    Article  CAS  PubMed  Google Scholar 

  2. Kasschau KD, Xie Z, Allen E, Llave C, Chapman EJ, Krizan KA, Carrington JC (2003) P1/HC-Pro, a viral suppressor of RNA silencing, interferes with Arabidopsis development and miRNA function. Dev Cell 4:205–217

    Article  CAS  PubMed  Google Scholar 

  3. Volpe TA, Kidner C, Hall I, Teng MG, Grewal SIS, Martienssen RA (2002) Regulation of heterochromatic silencing and histone H3 lysine-9 methylation by RNAi. Science 297:1833–1837

    Article  CAS  PubMed  Google Scholar 

  4. Jones L, Ratcliff F, Baulcombe DC (2001) RNA-directed transcriptional gene silencing in plants can be inherited independently of the RNA trigger and requires Met1 for maintenance. Curr Biol 11:747–757

    Article  CAS  PubMed  Google Scholar 

  5. Schiebel W, Haas B, Marinković S, Klanner A, Sänger HL (1993) RNA-directed RNA polymerase from tomato leaves. II. Catalytic in vitro properties. J Biol Chem 268:11858–11867

    CAS  PubMed  Google Scholar 

  6. Wassenegger M, Krczal G (2006) Nomenclature and functions of RNA-directed RNA polymerases. Trends Plant Sci 11:142–151

    Article  CAS  PubMed  Google Scholar 

  7. Astier-Manifacier S, Cornuet P (1971) RNA-dependent RNA polymerase in Chinese cabbage. Biochim Biophys Acta 232:484–493

    CAS  PubMed  Google Scholar 

  8. Yu D, Fan B, MacFarlane SA, Chen Z (2003) Analysis of the involvement of an inducible Arabidopsis RNA-dependent RNA polymerase in antiviral defense. Mol Plant-Microbe Interact 16:206–216

    Article  CAS  PubMed  Google Scholar 

  9. Pandey SP, Baldwin IT (2007) RNA-directed RNA polymerase 1 (RdR1) mediates the resistance of Nicotiana attenuata to herbivore attack in nature. Plant J 50:40–53

    Article  CAS  PubMed  Google Scholar 

  10. Xie ZX, Johansen LK, Gustafson AM, Kasschau KD, Lellis AD, Zilberman D, Jacobsen SE, Carrington JC (2004) Genetic and functional diversification of small RNA pathways in plants. PLoS Biol 2:642–652

    Article  CAS  Google Scholar 

  11. Alleman M, Sidorenko L, McGinnis K, Seshadri V, Dorweiler JE, White J, Sikkink K, Chandler VL (2006) An RNA-dependent RNA polymerase is required for paramutation in maize. Nature 442:295–298

    Article  CAS  PubMed  Google Scholar 

  12. Pandey SP, Baldwin IT (2008) Silencing RNA-directed RNA polymerase 2 increases the susceptibility of Nicotiana attenuata to UV in the field and in the glasshouse. Plant J 54:845–862

    Article  CAS  PubMed  Google Scholar 

  13. Mourrain P, Beclin C, Elmayan T, Feuerbach F, Godon C, Morel JB, Jouette D, Lacombe AM, Nikic S, Picault N et al (2000) Arabidopsis SGS2 and SGS3 genes are required for posttranscriptional gene silencing and natural virus resistance. Cell 101:533–542

    Article  CAS  PubMed  Google Scholar 

  14. Peragine A, Yoshikawa M, Wu G, Albrecht HL, Poethig RS (2004) SGS3 and SGS2/SDE1/RDR6 are required for juvenile development and the production of trans-acting siRNAs in Arabidopsis. Genes Dev 18:2368–2379

    Article  CAS  PubMed  Google Scholar 

  15. Vazquez F, Vaucheret H, Rajagopalan R, Lepers C, Gasciolli V, Mallory AC, Hilbert JL, Bartel DP, Crete P (2004) Endogeneous trans-acting siRNAs regulate the accumulation of Arabidopsis mRNAs. Mol Cell 16:69–79

    Article  CAS  PubMed  Google Scholar 

  16. Borsani O, Zhu J, Verslues PE, Sunkar R, Zhu JK (2005) Endogeneous siRNAs derived from a pair of natural cis-antisense transcripts regulate salt tolerance in Arabidopsis. Cell 123:1279–1291

    Article  CAS  PubMed  Google Scholar 

  17. Pandey SP, Gaquerel E, Gase K, Baldwin IT (2008) RNA-directed RNA polymerase 3 from Nicotiana attenuata is required for competitive growth in natural environments. Plant Physiol 147:1212–1224

    Article  CAS  PubMed  Google Scholar 

  18. Wang J, Wang X, Liu C, Zhang J, Zhu C, Guo X (2008) The NgAOX1a gene cloned from Nicotiana glutinosa is implicated in the response to abiotic and biotic stresses. Biosci Rep 28:259–266

    Article  PubMed  Google Scholar 

  19. Lorkovic ZJ, Wieczorek Kirk DA, Lambermon MH, Filipowicz W (2000) Pre-mRNA splicing in higher plants. Trends Plant Sci 5:160–167

    Article  CAS  PubMed  Google Scholar 

  20. Yang JH, Seo HH, Han SJ, Yoon EK, Yang MS, Lee WS (2008) Phytohormone abscisic acid control RNA-dependent RNA polymerase 6 gene expression and post-transcriptional gene silencing in rice cells. Nucleic Acids Res 36:1220–1226

    Article  CAS  PubMed  Google Scholar 

  21. Dalmay T, Hamilton A, Rudd S, Angell S, Baulcombe DC (2000) An RNA-dependent RNA polymerase gene in Arabidopsis is required for posttranscriptional gene silencing mediated by a transgene but not by a virus. Cell 101:543–553

    Article  CAS  PubMed  Google Scholar 

  22. Qu F, Ye X, Hou G, Sato S, Clemente TE, Morris TJ (2005) RDR6 has a broad-spectrum but temperature-dependent antiviral defense role in Nicotiana benthamiana. J Virol 79:15209–15217

    Article  CAS  PubMed  Google Scholar 

  23. Voinnet O (2001) RNA silencing as a plant immune system against viruses. Trends Genet 17:449–459

    Article  CAS  PubMed  Google Scholar 

  24. Klessig DF, Durner J, Noad R, Navarre DA, Wendehenne D, Kumar D, Zhou JM, Shah J, Zhang S, Kachroo P, Trifa Y, Pontier D, Lam E, Silva H (2000) Nitric oxide and salicylic acid signaling in plant defense. Proc Natl Acad Sci USA 97:8849–8855

    Article  CAS  PubMed  Google Scholar 

  25. Dong X (1998) SA, JA, ethylene, and disease resistance in plants. Curr Opin Plant Biol 1:316–323

    Article  CAS  PubMed  Google Scholar 

  26. Adie BA, Pérez-Pérez J, Pérez-Pérez MM, Godoy M, Sánchez-Serrano JJ, Schmelz EA, Solano R (2007) ABA is an essential signal for plant resistance to pathogens affecting JA biosynthesis and the activation of defenses in Arabidopsis. Plant Cell 19:1665–1681

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was financially supported in part by the National Natural Science Foundation of China (No. 30571215 and No. 30771408). We also want to express our gratitude to Dr. Xue-Shui Guo (Department of Biology and Microbiology, South Dakota State University. Brookings, SD. USA.) for critical reading of the manuscript.

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Changxiang Zhu or Xingqi Guo.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yang, H., Wang, M., Gao, Z. et al. Isolation of a novel RNA-dependent RNA polymerase 6 from Nicotiana glutinosa, NgRDR6, and analysis of its response to biotic and abiotic stresses. Mol Biol Rep 38, 929–937 (2011). https://doi.org/10.1007/s11033-010-0186-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-010-0186-z

Keywords

Navigation