Skip to main content

Advertisement

SpringerLink
Log in

Cloning and characterization of a novel secretory root-expressed peroxidase gene from common bean (Phaseolus vulgaris L.) infected with Fusarium oxysporum f. sp. Phaseoli

  • Published:
Molecular Breeding Aims and scope Submit manuscript

Abstract

Peroxidases play an important role in plant defense response to the attack of a host tissue by many pathogens. In this study, a new POX gene was isolated from common bean roots using cDNA-AFLP libraries and differential display of tissues with and without Fusarium oxysporum f. sp. phaseoli infection. Rapid amplification of cDNA ends was used to clone the full gene. The source of the differentially expressed fragment was the resistant genotype CAAS260205. The common bean POX gene showed a similar gene structure to a soybean POX gene consisting of four exons and three introns. Sequence analysis showed that PvPOX1 contained two conserved functional domains: a peroxidae active enzymatic site and a peroxidase heme-ligand site in close proximity. Phylogenetic study differentiated the new PvPOX1 gene from previous peroxidases in this multi-gene family. Real-time PCR analyses indicated that expression of the PvPOX1 gene was upregulated by Fusarium infection and that peroxidase activity, as measured by hydrogen peroxide (H2O2) accumulation, was enhanced during disease. The mRNA transcript level of PvPOX1 was also significantly increased following the stresses of wounding and polyethylene glycol, salicylic acid and abscisic acid treatments. The results of subcellular localization studies indicated that the localization of the PvPOX1 protein was in the apoplastic region of root cells. A co-dominant marker was developed for genetic mapping of the PvPOX1 gene based on an indel/insertion between the sequences of Andean and Mesoamerican parental genotypes (Hongyundou and Jingdou), which are mapping parents used in resistance breeding. Based on the new genetic marker, the gene was located between simple sequence repeat markers BMg2113 and BM181 on chromosome 3 of common bean and was not related to other peroxidases in terms of location. Overall, these results facilitate understanding of the molecular mechanism controlling resistance to Fusarium wilt pathogens and provide a diagnostic marker for selection of resistance to this disease based on peroxidase expression in common bean.

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
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Abl AJ, Guest DI, Sutherland MW (2000) Hydrogen peroxide yields during the incompatible interaction of tobacco suspension cells inoculated with Phytophthora nicotianae. Plant Physiol 124:899–910

    Article  Google Scholar 

  • Almagro L, Gómez Ros LV, Belchi-Navarro S, Bru R, Ros Barceló A, Pedreño MA (2009) Class III peroxidases in plant defence reactions. J Exp Bot 60:377–390

    Article  PubMed  CAS  Google Scholar 

  • Anderson JP, Badruzsaufari E, Schenk PM, Manners JM, Desmond OJ, Ehlert C, Maclean DJ, Ebert PR, Kazan K (2004) Antagonistic interaction between abscisicacid and jasmonate-ethylene signaling pathways modulates defense gene expression and disease resistance in Arabidopsis. Plant Cell 16:3460–3479

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Bachem CWB, van der Hoeven RS, de Bruijn SM, Vreugdenhil D, Zabeau M, Visser RG (1996) Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: analysis of gene expression during potato tuber development. Plant J 9:745–753

    Article  PubMed  CAS  Google Scholar 

  • Bakalovic N, Passardi F, Ioannidis V, Cosio C, Penel C, Falquet L, Dunand C (2006) PeroxiBase: a class III plant peroxidase database. Phytochemistry 67:534–539

    Article  PubMed  CAS  Google Scholar 

  • Bassam BJ, Caetano-Anollés G, Gresshoff PM (1991) Fast and sensitive silver staining of DNA in polyacrylamide gels. Anal Biochem 196:80–83

    Article  PubMed  CAS  Google Scholar 

  • Berrocal-Lobo M, Molina A (2008) Arabidopsis defense response against Fusarium oxysporum. Trends Plant Sci 13:145–150

    Article  PubMed  CAS  Google Scholar 

  • Blair MW, Pedraza F, Buendia HF, Gaitán-Solís E, Beebe SE, Gepts P, Tohme J (2003) Development of a genome-wide anchored microsatellite map for common bean (Phaseolus vulgaris L.). Theor Appl Genet 107:1362–1374

    Article  PubMed  CAS  Google Scholar 

  • Blair MW, Giraldo MC, Buendia HF, Tovar E, Duque MC, Beebe SE (2006) Microsatellite marker diversity in common bean (Phaseolus vulgaris L.). Theor Appl Genet 113:100–109

    Article  PubMed  CAS  Google Scholar 

  • Blair MW, Díaz LM, Buendía HF, Duque MC (2009) Genetic diversity, seed size associations and population structure of a core collection of common beans (Phaseolus vulgaris L.). Theor Appl Genet 119:955–973

    Article  PubMed  CAS  Google Scholar 

  • Blee KA, Jupe SC, Richard G, Zimmerlin A, Davies DR, Bolwell GP (2001) Molecular identification and expression of the peroxidase responsible for the oxidative burst in French bean (Phaseolus vulgaris L.) and related members of the gene family. Plant Mol Biol 47:607–620

    Article  PubMed  CAS  Google Scholar 

  • Bolwell GP, Bindschedler LV, Blee KA, Butt VS, Davies DR, Gardner SL, Gerrish C, Minibayeva F (2002) The apoplastic oxidative burst in response to biotic stress in plants: a three-component system. J Exp Bot 53:1367–1376

    Article  PubMed  CAS  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 Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  • Chen JB, Wang SM, Jing RL, Mao XG (2009) Cloning the PvP5CS gene from common bean (Phaseolus vulgaris) and its expression patterns under abiotic stresses. J Plant Physiol 166:12–19

    Article  PubMed  CAS  Google Scholar 

  • Chen ML, Wu J, Wang LF, Blair MW, Wang SM (2013) Development of mapped simple sequence repeat markers from common bean (Phaseolus vulgaris L.) based on genome sequences of a Chinese landrace and diversity evaluation. Mol Breed. doi:10.1007/s11032-013-9949-2

  • Cheong YH, Chang HS, Gupta R, Wang X, Zhu T, Luan S (2002) Transcriptional profiling reveals novel interactions between wounding, pathogen, abiotic stress, and hormonal responses in Arabidopsis. Plant Physiol 129:661–677

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Collinge M, Boller T (2001) Differential induction of two potato genes, Stprx2 and StNAC, in response to infection by Phytophthora infestans and to wounding. Plant Mol Biol 46:521–529

    Article  PubMed  CAS  Google Scholar 

  • Costa MM, Hilliou F, Duarte P, Pereira LG, Almeida I, Leech M, Memelink J, Barceló AR, Sottomayor M (2008) Molecular cloning and characterization of a vacuolar class III peroxidase involved in the metabolism of anticancer alkaloids in Catharanthus roseus. Plant Physiol 146:403–417

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Curtis A, Wilkinson C (1997) Topographical control of cells. Biomaterials 18:1573–1583

    Article  PubMed  CAS  Google Scholar 

  • Do HM, Hong JK, Jung HW, Kim SH, Ham JH, Hwang BK (2003) Expression of peroxidase-like genes, H2O2 production, and peroxidase activity during the hypersensitive response to Xanthomonas campestris pv. vesicatoria in Capsicum annuum. Mol Plant Microbe Interact 16:196–205

    Article  PubMed  CAS  Google Scholar 

  • Fall AL, Byrne PF, Jung G, Coyne DP, Brick MA, Schwartz HF (2001) Detection and mapping of a major mocus for fusarium wilt resistance in common bean. Crop Sci 41:1494–1498

    Article  Google Scholar 

  • Hiraga S, Sasaki K, Ito H, Ohashi Y, Matsui H (2001) A large family of class III plant peroxidases. Plant Cell Physiol 42:462–468

    Article  PubMed  CAS  Google Scholar 

  • Jones JDG, Dangl JL (2006) The plant immune system. Nature 444:323–329

    Article  PubMed  CAS  Google Scholar 

  • Kumar S, Dutta A, Sinha AK, Sen J (2007) Cloning, characterization and localization of a novel basic peroxidase gene from Catharanthus roseus. FEBS J 274:1290–1303

    Article  PubMed  CAS  Google Scholar 

  • Lamb C, Dixon RA (1997) The oxidative burst in plant disease resistance. Annu Rev Plant Physiol Plant Mol Biol 48:251–275

    Article  PubMed  CAS  Google Scholar 

  • Liu G, Sheng X, Greenshields DL, Ogieglo A, Kaminskyj S, Selvaraj G, Wei Y (2005) Profiling of wheat class III peroxidase genes derived from powdery mildew-attacked epidermis reveals distinct sequence-associated expression patterns. Mol Plant Microbe Interact 18:730–741

    Article  PubMed  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408

    Article  PubMed  CAS  Google Scholar 

  • Mao X, Zhang H, Tian S, Chang X, Jing R (2010) TaSnRK2.4, an SNF1-type serine/threonine protein kinase of wheat (Triticum aestivum L.), confers enhanced multistress tolerance in Arabidopsis. J Exp Bot 61:683–696

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Melillo MT, Leonetti P, Bongiovanni M, Castagnone-Sereno P, Bleve-Zacheo T (2006) Modulation of reactive oxygen species activities and H2O2 accumulation during compatible and incompatible tomato-root-knot nematode interactions. New Phytol 170:501–512

    Article  PubMed  CAS  Google Scholar 

  • Miklas PN, Kelly JD, Beebe SE, Blair MW (2006) Common bean breeding for resistance against biotic and abiotic stresses: from classical to MAS breeding. Euphytica 147:105–131

    Article  CAS  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  PubMed  CAS  Google Scholar 

  • Passardi F, Longet D, Penel C, Dunand C (2004) The class III peroxidase multigenic family in rice and its evolution in land plants. Phytochemistry 65:1879–1893

    Article  PubMed  CAS  Google Scholar 

  • Passardi F, Cosio C, Penel C, Dunand C (2005) Peroxidases have more functions than a Swiss army knife. Plant Cell Rep 24:255–265

    Article  PubMed  CAS  Google Scholar 

  • Pastori GM, Foyer CH (2002) Common components, networks, and pathways of cross-tolerance to stress. The central role of “redox” and abscisic acid-mediated controls. Plant Physiol 129:460–468

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Ribeiro R, Hagedorn DJ (1979a) Screening for resistance to end pathogenic specialisation of Fusarium oxysporum f. sp. phaseoli, the causal agent of Bean Yellows. Phytopathology 69:272–276

  • Ribeiro R, Hagedorn DJ (1979b) Inheritance and nature of resistance in beans to Fusarium oxysporum f. sp. phaseoli. Phytopathology 69:859–861

  • Román-Avilés B, Kelly JD (2005) Identification of quantitative trait loci conditioning resistance to fusarium root rot in common bean. Crop Sci 45:1881–1890

    Article  Google Scholar 

  • Ryals JA, Neuenschwander UH, Willits MG, Molina A, Steiner HY, Hunt MD (1996) Systemic acquired resistance. Plant Cell 8:1809–1819

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Sagisaka S (1976) The occurrence of peroxide in a perennial plant, Populus gelrica. Plant Physiol 57:308–309

  • Schneider KA, Grafton KF, Kelly JD (2001) QTL analysis of resistance to fusrarium root rot in bean. Crop Sci 41:535–542

    Article  CAS  Google Scholar 

  • Simonetti E, Veronico P, Melillo MT, Delibes A, Andrés MF, López-Braña I (2009) Analysis of class III peroxidase genes expressed in roots of resistant and susceptible wheat lines infected by Heterodera avenae. Mol Plant Microbe Interact 22:1081–1092

    Article  PubMed  CAS  Google Scholar 

  • Summer H, Grämer R, Dröge P (2009) Denaturing urea polyacrylamide gel electrophoresis (Urea PAGE). J Vis Exp 32:1485

    PubMed  Google Scholar 

  • Van Loon LE, Strien Van (1999) The families of pathogenesis-related proteins, their activities, and comparative analysis of PR-1 type proteins. Physiol Mol Plant Pathol 55:85–97

    Article  Google Scholar 

  • Welinder KG, Ustesen AF, Kjaersgård IV, Jensen RB, Rasmussen SK, Jespersen HM, Duroux L (2002) Structural diversity and transcription of class III peroxidases from Arabidopsis thaliana. Eur J Biochem 269:6063–6081

    Article  PubMed  CAS  Google Scholar 

  • Xue RF, Zhu ZD, Huang Y, Wang XM, Wang LF, Wang SM (2012) Quantification of Fusarium oxysporum f. sp. phaseoli detected by real-time quantitative PCR in different common beans cultivars. Acta Agron Sin 38:791–799

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the Ministry of Agriculture of China [the earmarked fund for Modern Agro-industry Technology Research System (CARS-09-G12 and CARS-09-Z8)], National Key Technology Research and Development Program of the Ministry of Science and Technology of China (2013BAD01B03-18a) and the Scientific Innovation Program of CAAS. We thanked Dr. Li Zhensheng, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, for providing pJIT163-GFP expression vector. We also thank Dr. Li A. for excellent technical assistance and the instrumental services from Chinese Academy of Agricultural Sciences.

Author information

Authors and Affiliations

  1. National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Ren Feng Xue, Jing Wu, Ming Li Chen, Zhen Dong Zhu, Lan Fen Wang, Xiao Ming Wang & Shu Min Wang

  2. Crop Research Institute, Liaoning Academy of Agricultural Sciences, Shenyang, 110161, China

    Ren Feng Xue

  3. Department of Agriculture and Natural Sciences, Tennessee State University, Nashville, TN, 37209, USA

    Matthew W. Blair

Authors
  1. Ren Feng Xue
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jing Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ming Li Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zhen Dong Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lan Fen Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xiao Ming Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Matthew W. Blair
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Shu Min Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shu Min Wang.

Additional information

Ren Feng Xue and Jing Wu equally contributed to this article.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Xue, R.F., Wu, J., Chen, M.L. et al. Cloning and characterization of a novel secretory root-expressed peroxidase gene from common bean (Phaseolus vulgaris L.) infected with Fusarium oxysporum f. sp. Phaseoli . Mol Breeding 34, 855–870 (2014). https://doi.org/10.1007/s11032-014-0080-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11032-014-0080-9

Keywords

Search

Navigation