Skip to main content
SpringerLink
Log in

Decreased ROS level and activation of antioxidant gene expression in Agrobacterium rhizogenes pRiA4-transformed calli of Rubia cordifolia

  • Original Article
  • Published:
Planta Aims and scope Submit manuscript

Abstract

Microbe–plant interactions often lead to a decrease in the reactive oxygen species (ROS) level of plant cells, which allows pathogen survival through the suppression of plant immune responses. In the present investigation, we tested whether transformation of Rubia cordifolia cells by Agrobacterium rhizogenes had a similar effect. We isolated partial cDNA sequences of ascorbate peroxidase, catalase and Cu/Zn superoxide dismutase genes (RcApx1, RcApx2, RcApx3, RcCAT1, RcCAT2, RcCSD1, RcCSD2 and RcCSD3) from plant tissues, as well as pRiA4-transformed and normal calli of Rubia cordifolia, and studied their expression by real-time PCR. Transcription profiling revealed that ascorbate peroxidase (RcApx1) and Cu/Zn superoxide dismutase (RcCSD1) were the most abundant transcripts present in both plant tissues and non-transformed calli. Catalase genes were weakly expressed in these samples. The pRiA4-transformed calli showed enhanced expression of several genes encoding ROS-detoxifying enzymes. Confocal microscopy imaging revealed decreased ROS level in pRiA4-transformed calli compared to the control. These results demonstrate that A. rhizogenes, like other plant pathogens, uses a strategy aimed at decreasing ROS levels in host cells through the general upregulation of its antioxidant genes.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Abascal F, Zardoya R, Posada D (2005) ProtTest: selection of best-fit models of protein evolution. Bioinformatics 21:2104–2105

    Article  CAS  PubMed  Google Scholar 

  • Acevedo A, Williamson JD, Scandalios JG (1991) Photoregulation of the Cat2 and Cat3 catalase genes in pigmented and pigment-deficient maize: the circadian regulation of Cat3 is superimposed on its quasi-constitutive expression in maize leaves. Genetics 127:601–608

    CAS  PubMed  Google Scholar 

  • Alscher RG, Erturk N, Heath LS (2002) Role of superoxide dismutases (SODs) in controlling oxidative stress in plants. J Exp Bot 53:1331–1341

    Article  CAS  PubMed  Google Scholar 

  • Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    Article  CAS  PubMed  Google Scholar 

  • Ashraf M (2009) Biotechnological approach of improving plant salt tolerance using antioxidants as markers. Biotechnol Adv 27:84–93

    Article  CAS  PubMed  Google Scholar 

  • Bretz JR, Mock NM, Charity JC, Zeyad S, Baker CJ, Hutcheson SW (2003) A translocated protein tyrosine phosphatase of Pseudomonas syringae pv. tomato DC3000 modulates plant defence response to infection. Mol Microbiol 49:389–400

    Article  CAS  PubMed  Google Scholar 

  • Bulgakov VP, Tchernoded GK, Mischenko NP, Khodakovskaya MV, Glazunov VP, Radchenko SV, Zvereva EV, Fedoreyev SA, Zhuravlev YN (2002) Effect of salicylic acid, methyl jasmonate, ethephon and cantharidin on anthraquinone production by Rubia cordifolia callus cultures transformed with the rolB and rolC genes. J Biotechnol 97:213–221

    Article  CAS  PubMed  Google Scholar 

  • Bulgakov VP, Tchernoded GK, Mischenko NP, Shkryl YN, Glazunov VP, Fedoreyev SA, Zhuravlev YN (2003) Effects of Ca(2+) channel blockers and protein kinase/phosphatase inhibitors on growth and anthraquinone production in Rubia cordifolia callus cultures transformed by the rolB and rolC genes. Planta 217:349–355

    Article  CAS  PubMed  Google Scholar 

  • Bulgakov VP, Tchernoded GK, Mischenko NP, Shkryl YN, Fedoreyev SA, Zhuravlev YN (2004) The rolB and rolC genes activate synthesis of anthraquinones in Rubia cordifolia cells by mechanism independent of octadecanoid signaling pathway. Plant Sci 166:1069–1075

    Article  CAS  Google Scholar 

  • Bulgakov VP, Aminin DL, Shkryl YN, Gorpenchenko TY, Veremeichik GN, Dmitrenok PS, Zhuravlev YN (2008) Suppression of reactive oxygen species and enhanced stress tolerance in Rubia cordifolia cells expressing the rolC oncogene. Mol Plant Microbe Interact 21:1561–1570

    Article  CAS  PubMed  Google Scholar 

  • Crow JP (1997) Dichlorodihydrofluorescein and dihydrorhodamine 123 are sensitive indicators of peroxynitrite in vitro: implications for intracellular measurement of reactive nitrogen and oxygen species. Nitric Oxide 1:145–157

    Article  CAS  PubMed  Google Scholar 

  • Davletova S, Rizhsky L, Liang H, Shengqiang Z, Oliver DJ, Coutu J, Shulaev V, Schlauch K, Mittler R (2005) Cytosolic ascorbate peroxidase 1 is a central component of the reactive oxygen gene network of Arabidopsis. Plant Cell 17:268–281

    Article  CAS  PubMed  Google Scholar 

  • Du YY, Wang PC, Chen J, Song CP (2008) Comprehensive functional analysis of the catalase gene family in Arabidopsis thaliana. J Integr Plant Biol 50:1318–1326

    Article  CAS  PubMed  Google Scholar 

  • Escobar MA, Dandekar AM (2003) Agrobacterium tumefaciens as an agent of disease. Trends Plant Sci 8:380–386

    Article  CAS  PubMed  Google Scholar 

  • Felsenstein J (1989) PHYLIP—Phylogeny Inference Package (Version 3.2). Cladistics 5:164–166

    Google Scholar 

  • Frugoli JA, Zhong HH, Nuccio ML, McCourt P, McPeek MA, Thomas TL, McClung CR (1996) Catalase is encoded by a multigene family in Arabidopsis thaliana (L.) Heynh. Plant Physiol 112:327–336

    Article  CAS  PubMed  Google Scholar 

  • Gechev TS, Van Breusegem F, Stone JM, Denev I, Laloi C (2006) Reactive oxygen species as signals that modulate plant stress responses and programmed cell death. Bioessays 28:1091–1101

    Article  CAS  PubMed  Google Scholar 

  • Guindon S, Gascuel O (2003) A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol 52:696–704

    Article  PubMed  Google Scholar 

  • Hindges R, Slusarenko A (1992) cDNA and derived amino acid sequence of a cytosolic Cu, Zn superoxide dismutase from Arabidopsis thaliana (L.) Heynh. Plant Mol Biol 18:123–125

    Article  CAS  PubMed  Google Scholar 

  • Jeanmougin F, Thompson JD, Gouy M, Higgins M, Gibson TJ (1998) Multiple sequence alignment with ClustalX. Trends Biochem Sci 23:403–405

    Article  CAS  PubMed  Google Scholar 

  • Karpinski S, Reynolds H, Karpinska B, Wingsle G, Creissen G, Mullineaux P (1999) Systemic signaling and acclimation in response to excess excitation energy in Arabidopsis. Science 284:654–657

    Article  CAS  PubMed  Google Scholar 

  • Kim HJ, Kato N, Kim S, Triplett B (2008) Cu/Zn superoxide dismutases in developing cotton fibers: evidence for an extracellular form. Planta 228:281–292

    Article  CAS  PubMed  Google Scholar 

  • Kliebenstein DJ, Monde RA, Last RL (1998) Superoxide dismutase in Arabidopsis: an eclectic enzyme family with disparate regulation and protein localization. Plant Physiol 118:637–650

    Article  CAS  PubMed  Google Scholar 

  • Mittler R, Vanderauwera S, Gollery M, Van Breusegem F (2004) Reactive oxygen gene network of plants. Trends Plant Sci 9:490–498

    Article  CAS  PubMed  Google Scholar 

  • Narendra S, Venkataramani S, Shen G, Wang J, Pasapula V, Lin Y, Kornyeyev D, Holaday AS, Zhang H (2006) The Arabidopsis ascorbate peroxidase 3 is a peroxisomal membrane-bound antioxidant enzyme and is dispensable for Arabidopsis growth and development. J Exp Bot 57:3033–3042

    Article  CAS  PubMed  Google Scholar 

  • Page RD (1996) TreeView: an application to display phylogenetic trees on personal computers. Comput Appl Biosci 12:357–358

    CAS  PubMed  Google Scholar 

  • Panchuk II, Volkov RA, Schoffl F (2002) Heat stress- and heat shock transcription factor-dependent expression and activity of ascorbate peroxidase in Arabidopsis. Plant Physiol 129:838–853

    Article  CAS  PubMed  Google Scholar 

  • Robinette D, Matthysse AG (1990) Inhibition by Agrobacterium tumefaciens and Pseudomonas savastanoi of development of the hypersensitive response elicited by Pseudomonas syringae pv. phaseolicola. J Bacteriol 172:5742–5749

    CAS  PubMed  Google Scholar 

  • Shigeoka S, Ishikawa T, Tamoi M, Miyagawa Y, Takeda T, Yabuta Y, Yoshimura K (2002) Regulation and function of ascorbate peroxidase isoenzymes. J Exp Bot 53:1305–1319

    Article  CAS  PubMed  Google Scholar 

  • Shkryl YN, Veremeichik GN, Bulgakov VP, Tchernoded GK, Mischenko NP, Fedoreyev SA, Zhuravlev YN (2008) Individual and combined effects of the rolA, B, and C genes on anthraquinone production in Rubia cordifolia transformed calli. Biotechnol Bioeng 100:118–125

    Article  CAS  PubMed  Google Scholar 

  • Slightom J, Durand-Tardif M, Joanin L, Tepfer D (1986) Nucleotide sequence analysis of TL-DNA of Agrobacterium rhizogenes agropine type plasmid. J Biol Chem 261:108–121

    CAS  PubMed  Google Scholar 

  • Teixeira FK, Menezes-Benavente L, Galvão VC, Margis R, Margis-Pinheiro M (2006) Rice ascorbate peroxidase gene family encodes functionally diverse isoforms localized in different subcellular compartments. Planta 224:300–314

    Article  CAS  PubMed  Google Scholar 

  • Torres MA, Jones JD, Dangl JL (2006) Reactive oxygen species signaling in response to pathogens. Plant Physiol 141:373–378

    Article  CAS  PubMed  Google Scholar 

  • Underwood W, Zhang S, He SY (2007) The Pseudomonas syringae type III effector tyrosine phosphatase HopAO1 suppresses innate immunity in Arabidopsis thaliana. Plant J 52:658–672

    Article  CAS  PubMed  Google Scholar 

  • Xiong L, Schumaker KS, Zhu J-K (2002) Cell signaling during cold, drought, and salt stress. Plant Cell 14:165–183

    Article  Google Scholar 

  • Xu XQ, Pan SQ (2000) An Agrobacterium catalase is a virulence factor involved in tumorigenesis. Mol Microbiol 35:407–414

    Article  CAS  PubMed  Google Scholar 

  • Zhong HH, McClung CR (1996) The circadian clock gates expression of two Arabidopsis catalase genes to distinct and opposite circadian phases. Mol Gen Genet 251:196–203

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the Russian Foundation for Basic Research, by the Grant Program “Molecular and Cell Biology” of the Russian Academy of Sciences, and by grants from the Far East Branch of the Russian Academy of Sciences 09-III-A-06-187, 10-III-B-06-086, 09-III-A-05-137. The authors would like to express their gratitude to Dr. S. Shedko, Dr. E. Artyukova, Dr. M. Kozyrenko and Dr. A. Gontcharov for their help with phylogenetic analysis.

Author information

Authors and Affiliations

  1. Institute of Biology and Soil Science, Far East Branch of Russian Academy of Sciences, 159 Stoletija Street, Vladivostok, 690022, Russia

    Y. N. Shkryl, G. N. Veremeichik, V. P. Bulgakov, T. Y. Gorpenchenko & Y. N. Zhuravlev

  2. Pacific Institute of Bioorganic Chemistry, Far East Branch of Russian Academy of Sciences, 159 Stoletija Street, Vladivostok, 690022, Russia

    D. L. Aminin

Authors
  1. Y. N. Shkryl
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G. N. Veremeichik
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. V. P. Bulgakov
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. T. Y. Gorpenchenko
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. D. L. Aminin
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Y. N. Zhuravlev
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Y. N. Shkryl.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Suppl. Fig. 1

. Maximum-likelihood cladogram for ascorbate peroxidase isoenzymes obtained using Phyml program (300 bootstraps). Similar tree was also obtained with the maximum parsimony method (from Phylip package, 1,000 bootstraps). Numbers on the branches represent bootstrap values as obtained from ML/MP, respectively (only values >50% are presented; lower values are denoted ‘-’). The GenBank accession numbers of the proteins are given in the cladogram. The sequences obtained in this work are marked by dots (JPG 204 kb)

Suppl. Fig. 2

. Maximum-likelihood cladogram for Cu/Zn superoxide dismutase isoenzymes obtained using Phyml program (300 bootstraps). Similar tree was also obtained with the maximum parsimony method (from Phylip package, 1,000 bootstraps). Numbers on the branches represent bootstrap values as obtained from ML/MP, respectively (only values >50% are presented; lower values are denoted ‘-’). The GenBank accession numbers of the proteins are given in the cladogram. The sequences obtained in this work are marked by dots (JPG 146 kb)

Suppl. Fig. 3

. Maximum-likelihood cladogram for catalase isoenzymes obtained using Phyml program (300 bootstraps). Similar tree was also obtained with the maximum parsimony method (from Phylip package, 1,000 bootstraps). Numbers on the branches represent bootstrap values as obtained from ML/MP, respectively (only values >50% are presented; lower values are denoted ‘-’). The GenBank accession numbers of the proteins are given in the cladogram. The sequences obtained in this work are marked by dots (JPG 144 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shkryl, Y.N., Veremeichik, G.N., Bulgakov, V.P. et al. Decreased ROS level and activation of antioxidant gene expression in Agrobacterium rhizogenes pRiA4-transformed calli of Rubia cordifolia . Planta 232, 1023–1032 (2010). https://doi.org/10.1007/s00425-010-1237-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-010-1237-3

Keywords

Search

Navigation