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Analysis of ascorbate peroxidase genes expressed in resistant and susceptible wheat lines infected by the cereal cyst nematode, Heterodera avenae

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Abstract

Changes in ascorbate peroxidase (APX) enzyme activity in response to nematode (Heterodera avenae) attack were studied in roots of three hexaploid wheat lines carrying Cre2, Cre5, or Cre7 nematode resistance genes and the susceptible Triticum aestivum cv. Anza. A spectrophotometric analysis was carried out with root extracts of infected plants 4, 7, 11, and 14 days after nematode inoculation using uninfected plant as control. APX induction in infected resistant genotypes was similar and higher than in the susceptible control. The introgression wheat/Aegilops ventricosa H-93-8 line, carrying the Cre2 gene, and its parental line H-10-15 as susceptible control were used to analyze whether this increase of activity was correlated with the induction of APX gene expression. Genes encoding cytosolic forms of APX were induced in roots of both lines in response to nematode infection. This induction took place both earlier and with greater intensity in the resistant line than in the susceptible one, and it was also higher in the root area at the site of nematode attachment.

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Abbreviations

APX:

Ascorbate peroxidase

AsA:

Ascorbic acid

Cre:

Cereal root eelworm

CCN:

Cereal cyst nematode

dai:

Days after inoculation

EST:

Esterase

GSH:

Reduced form of glutathione

PRX:

Plant class III peroxidases

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

References

  • Alkharouf N, Khan R, Matthews BF (2004) Analysis of expressed sequence tags from roots of resistant soybean infected by the soybean cyst nematode. Genome 47:380–388

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Andrés MF, Melillo MT, Delibes A, Romero MD, Bleve-Zacheo T (2001) Changes in wheat root enzymes correlated with resistance to cereal cyst nematodes. New Phytol 152:343–354

    Article  Google Scholar 

  • Båga M, Chibbar RN, Kartha KK (1995) Molecular cloning and expression analysis of peroxidase genes from wheat. Plant Mol Biol 29:647–662

    Article  PubMed  Google Scholar 

  • Bai R, Ma F, Liang D, Zhao X (2009) Phthalic acid induces oxidative stress and alters the activity of some antioxidant enzymes in roots of Malus prunifolia. J Chem Ecol 35:488–494

    Article  CAS  PubMed  Google Scholar 

  • Barcala M, García A, Cabrera J, Casson S, Lindsey K, Favery B, García-Casado G, Solano R, Fenoll C, Escobar C (2010) Early transcriptomic events in microdissected Arabidopsis nematode-induced giant cells. Plant J 61:698–712

    Article  CAS  PubMed  Google Scholar 

  • Bindschedler LV, Dewdney J, Blee KA, Stone JM, Asai T, Plotnikov J, Denoux C, Hayes T, Gerrish C, Davies DR, Ausubel FM, Bolwell GP (2006) Peroxidase-dependent apoplastic oxidative burst in Arabidopsis required for pathogen resistance. Plant J 47:851–863

    Article  CAS  PubMed  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  CAS  PubMed  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for 1 the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Chaudie`re J, Ferrari-Iliou R (1999) Intracellular antioxidants: from chemical to biochemical mechanisms. Food Chem Toxicol 37:949–962

    Article  CAS  Google Scholar 

  • Chen GX, Asada K (1989) Ascorbate peroxidase in tea leaves: occurrence of two isoenzymes and the differences in their enzymatic and molecular properties. Plant Cell Physiol 30:987–998

    CAS  Google Scholar 

  • Chen YP, Wang HZ, Cao AZ, Wang CM, Chen PD (2006) Cloning of a resistance gene analog from wheat and development of a codominant PCR marker for Pm21. J Integr Plant Biol 48:715–721

    Article  CAS  Google Scholar 

  • Chew O, Whelan J, Millar AH (2003) Molecular definition of the ascorbate glutathione cycle in Arabidopsis mitochondria reveals dual targeting of antioxidant defenses in plants. J Biol Chem 278:46869–46877

    Article  CAS  PubMed  Google Scholar 

  • Corpet F (1988) Multiple sequence alignment with hierarchical clustering. Nucleic Acids Res 16:10881–10890

    Article  CAS  PubMed  Google Scholar 

  • Cosio C, Dunand C (2009) Specific functions of individual class III peroxidase genes. J Exp Bot 60:391–408

    Article  CAS  PubMed  Google Scholar 

  • Cosio C, Vuillemin L, De Meyer M, Kevers C, Penel C, Dunand C (2009) An anionic class III peroxidase from zucchini may regulate hypocotyl elongation through auxin oxidase activity. Planta 229:823–836

    Article  CAS  PubMed  Google Scholar 

  • Das S, DeMason DA, Ehlers JD, Close TJ, Roberts PA (2008) Histological characterization of root-knot nematode resistance in cowpea and its relation to reactive oxygen species modulation. J Exp Bot 59:1305–1313

    Article  CAS  PubMed  Google Scholar 

  • Delibes A, Romero D, Aguaded S, Duce A, Mena M, Lopez-Braña I, Andrés MF, Martin-Sanchez JA, García-Olmedo F (1993) Resistance to the cereal cyst nematode (Heterodera avenae Woll.) transferred from the wild grass Aegilops ventricosa to hexaploid wheat by a “stepping-stone procedure. Theor Appl Genet 87:402–408

    Article  Google Scholar 

  • Eastwood RF, Lagudah ES, Appels R (1994) A directed search for DNA sequence tightly linked to cereal cyst nematode resistance genes in Triticum tauschii. Genome 37:311–319

    Article  CAS  PubMed  Google Scholar 

  • Golinowski W, Grundler FMW, Sobczak M (1996) Changes in the structure of Arabidopsis thaliana during female development of the plant parasitic nematode Heterodera schachtii. Protoplasma 194:103–116

    Article  Google Scholar 

  • Ioannidi E, Kalamaki MS, Engineer C, Pateraki I, Alexandrou D, Mellidou I, Giovannonni J, Kanellis AK (2009) Expression profiling of ascorbic acid-related genes during tomato fruit development and ripening and in response to stress conditions. J Exp Bot 60:663–678

    Article  CAS  PubMed  Google Scholar 

  • Jahier J, Abelard P, Tanguy M, Dedryver F, Rivoal R, Khatkar S, Bariana HS (2001) The Aegilops ventricosa segment on chromosome 2AS of the wheat cultivar ‘VPM1’ carries the cereal cyst nematode resistance gene Cre5. Plant Breed 120:125–128

    Article  CAS  Google Scholar 

  • Johrde A, Schweizer P (2008) A class III peroxidase specifically expressed in pathogen-attacked barley epidermis contributes to basal resistance. Mol Plant Pathol 9:687–696

    Article  CAS  PubMed  Google Scholar 

  • Kawano T (2003) Roles of the reactive oxygen species-generating peroxidase reactions in plant defense and growth induction. Plant Cell Rep 21:829–837

    CAS  PubMed  Google Scholar 

  • Liu XM, Williams CE, Nemacheck JA, Wang H, Subramanyam S, Zheng C, Chen MS (2010) Reactive oxygen species are involved in plant defense against a gall midge. Plant Physiol 152:985–999

    Article  CAS  PubMed  Google Scholar 

  • López-Braña I, Romero MD, Delibes A (1996) Application of the random amplified polymorphic DNA (RAPD) technique for analysis of Heterodera avenae complex populations. Genome 39:118–122

    Article  PubMed  Google Scholar 

  • Mena M, Orellana J, López-Braña I, García-Olmedo F, Delibes A (1993) Characterization of wheat/Aegilops ventricosa introgression and addition lines with respect to the Mv genome. Theor Appl Genet 86:197–204

    Article  CAS  Google Scholar 

  • Mittler M, Zilinskas BA (1994) Regulation of pea cytosolic ascorbate peroxidase and other antioxidant enzymes during the progression of drought stress and following recovery from drought. Plant J 5:397–405

    Article  CAS  PubMed  Google Scholar 

  • Mittler R, Feng X, Cohen M (1998) Post-transcriptional suppression of cytosolic ascorbate peroxidase expression during pathogen-induced programmed cell death in tobacco. Plant Cell 10:461–474

    Article  CAS  PubMed  Google Scholar 

  • Montes MJ, Lopez-Braña I, Romero MD, Sin E, Andres MF, Martin-Sánchez JA, Delibes A (2003) Biochemical and genetic studies of two Heterodera avenae resistance genes transferred from Aegilops ventricosa to wheat. Theor Appl Genet 107:611–618

    Article  CAS  PubMed  Google Scholar 

  • Montes MJ, López-Braña I, Delibes A (2004) Root enzyme activities associated with resistance to Heterodera avenae conferred by gene Cre7 in a wheat/Aegilops triuncialis introgression line. J Plant Physiol 161:1135–1140

    Article  Google Scholar 

  • Montes MJ, Andrés MF, Sin E, López-Braña I, Martín-Sánchez JA, Romero MD, Delibes A (2008) Cereal cyst nematode resistance conferred by the Cre7 gene from Aegilops triuncialis and its relationship with Cre genes from Australian wheat cultivars. Genome 51:315–319

    Article  CAS  PubMed  Google Scholar 

  • Mullineaux P, Karpinski S (2000) Signal transduction in response to excess light: getting out of the chloroplast. Curr Opin Plant Biol 5:43–48

    Article  Google Scholar 

  • Nakano Y, Asada K (1981) Hydrogen-peroxide is scavenged by ascorbate specific peroxidase in spinach-chloroplasts. Plant Cell Physiol 22:867–880

    CAS  Google Scholar 

  • Nicol JM, Rivoal R (2007) Global knowledge and its application for the integrated control and management of nematodes on wheat. In: Ciancio A, Mukerji KG (eds) Integrated management and biocontrol of vegetable and grain crops nematodes. Springer, The Netherlands, pp 243–287

  • Nürnberger T, Brunner F, Kemmerling B, Piater L (2004) Innate immunity in plants and animals: striking similarities and obvious differences. Immunol Rev 198:249–266

    Article  PubMed  Google Scholar 

  • Ogbonnaya FC, Seah S, Delibes A, Jahier J, López-Braña I, Eastwood RF, Lagudah ES (2001) Molecular–genetic characterisation of a new nematode resistance gene in wheat. Theor Appl Genet 102:623–629

    Article  CAS  Google Scholar 

  • Orvar BL, Ellis BE (1997) Transgenic tobacco plants expressing antisense RNA for cytosolic ascorbate peroxidase show increased susceptibility to ozone injury. Plant J 11:1297–1305

    Article  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  CAS  PubMed  Google Scholar 

  • Passardi F, Theiler G, Zamocky M, Cosio C, Rouhier N, Teixeira F, Margis-Pinheiro M, Ioannidis V, Penel C, Falquet L, Dunand C (2007) Peroxibase: the peroxidase database. Phytochemistry 68:1605–1611

    Article  CAS  PubMed  Google Scholar 

  • Pereira CS, Soares da Costa D, Teixeira J, Pereira S (2005) Organ-specific distribution and subcellular localisation of ascorbate peroxidase isoenzymes in potato (Solanum tuberosum L.) plants. Protoplasma 226:223–230

    Article  CAS  PubMed  Google Scholar 

  • Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:e45

    Article  CAS  PubMed  Google Scholar 

  • Pnuelli L, Liang H, Rozenberg M, Mittler R (2003) Growth suppression, altered stomatal responses, and augmented induction of heat shock proteins in cytosolic ascorbate peroxidase (APX1)-deficient Arabidopsis plants. Plant J 34:187–203

    Article  Google Scholar 

  • Portillo M, Lindsey K, Casson S, García-Casado G, Solano R, Fenoll C, Escobar C (2009) Isolation of RNA from laser-capture microdissected giant cells at early differentiation stages suitable for differential transcriptome análisis. Mol Plant Pathol 10:523–535

    Article  CAS  PubMed  Google Scholar 

  • Rivoal R, Bekal S, Valette S, Gauthier JP, Bel Hadj Fradj M, Mokabli A, Jahier J, Nicol J, Yahyaoui A (2001) Variation in reproductive capacity and virulence on different genotypes and resistance genes of Triticeae in the cereal cyst nematode species complex. Nematology 3:581–592

    Article  Google Scholar 

  • Rizhsky L, Davletova S, Liang H, Mittler R (2004) The zinc finger protein Zat12 is required for cytosolic ascorbate peroxidase 1 expression during oxidative stress in Arabidopsis. J Biol Chem 279:11736–11743

    Article  CAS  PubMed  Google Scholar 

  • Romero D, Montes MJ, Sin E, López-Braña I, Duce A, Martín-Sánchez JA, Andrés MF, Delibes A (1998) A cereal cyst nematode (Heterodera avenae Woll.) resistance gene transferred from Aegilops triuncialis to hexaploid wheat. Theor App Genet 96:1135–1140

    Article  Google Scholar 

  • Safari E, Gororo NN, Eastwood RF, Lewis J, Eagles HA, Ogbonnaya FC (2005) Impact of Cre1, Cre8 and Cre3 genes on cereal cyst nematode resistance in wheat. Theor Appl Genet 110:567–572

    Article  CAS  PubMed  Google Scholar 

  • Schweizer P (2008) Tissue-specific expression of a defense-related peroxidase in transgenic wheat potentiates cell death in pathogen-attacked leaf epidermis. Mol Plant Pathol 9:45–57

    Article  CAS  PubMed  Google Scholar 

  • Seah S, Miller C, Sivasithamparam K, Lagudah ES (2000) Root responses to cereal cyst nematode (Heterodera avenae) in hosts with different resistance genes. New Phytol 146:527–533

    Article  Google Scholar 

  • Seckin B, Sekmen AH, Türkan I (2009) An enhancing effect of exogenous mannitol on the antioxidant enzyme activities in roots of wheat under salt stress. J Plant Growth Regul 28:12–20

    Article  CAS  Google Scholar 

  • Shi WM, Muramoto Y, Ueda A, Takabe T (2001) Cloning of peroxisomal ascorbate peroxidase gene from barley and enhanced thermotolerance by overexpressing in Arabidopsis thaliana. Gene 273:23–27

    Article  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 372:1305–1319

    Article  Google Scholar 

  • Simonetti E, Veronico P, Melillo MT, Delibes A, Andrés MF, López-Braña I (2009a) 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  CAS  PubMed  Google Scholar 

  • Simonetti E, Veronico P, Melillo MT, Bleve-Zacheo T, Delibes A, Andrés MF, López-Braña I (2009b) Class III peroxidase are involved in wheat resistance to Heterodera avenae. NEMAGENICS. Exploiting genomics to understand plant–nematode interactions. Cost 872 Workshop and Meeting. Toledo, Spain, p p 60 (abstracts book)

    Google Scholar 

  • Sobczak M, Avrova A, Jupowicz J, Phillips MS, Ernst K, Kumar A (2005) Characterization of susceptibility and resistance responses of potato cyst nematode (Globodera spp.) infection of tomato lines in the absence and presence of the broad spectrum nematode resistance Hero gene. Mol Plant–Microbe Interact 18:158–168

    Article  CAS  PubMed  Google Scholar 

  • Szakasits D, Heinen P, Wieczorek K, Hofmann J, Wagner F, Kreil DP, Sykacek P, Grundler FMW, Bohlmann H (2009) The transcriptome of syncytia induced by the cyst nematode Heterodera schachtii in Arabidopsis roots. Plant J 57:771–784

    Article  CAS  PubMed  Google Scholar 

  • Teixeira FK, Menezes-Benavente L, Margis R, Margis-Pinheiro M (2004) Analysis of the molecular evolutionary history of the ascorbate peroxidase gene family: inferences from the rice genome. J Mol Evol 59:761–770

    Article  CAS  PubMed  Google Scholar 

  • Teixeira FK, Menezes-Benavente L, Costa Galvão V, 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 

  • Welinder KG (1992) Plant peroxidases: structure-function relationships. In: Penel C et al. (eds) Plant peroxidases. Geneve University, Switzerland, pp 1–24

  • Zhao S, Fernald RD (2005) Comprehensive algorithm for quantitative real-time polymerase chain reaction. J Comp Biol 12:1047–1064

    Article  CAS  Google Scholar 

  • Zhao J, Davis LC, Verpoorte R (2005) Elicitor signal transduction leading to production of plant secondary metabolites. Biotechnol Adv 23:283–333

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This research was supported by Grant 083/Q060210-031 from the Comunidad Autonoma de Madrid (Spain). We thank M. López, C. Martínez-Belinchón and R. Rodriguez-Ríos for their technical assistance.

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  1. Ester Simonetti & Isidoro López-Braña

    Present address: Instituto de Investigaciones en Biociencias Agrícolas y Ambientales INBA-CONICET, Facultad de Agronomía, UBA, CABA, Buenos Aires, C1417DSE, Argentina

Authors and Affiliations

  1. Departamento de Biotecnología, ETS Ing. Agrónomos, UPM, Avda Complutense s/n, Madrid, 28040, Spain

    Ester Simonetti, Eva Alba, María Jesús Montes, Ángeles Delibes & Isidoro López-Braña

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  1. Ester Simonetti
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  2. Eva Alba
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  3. María Jesús Montes
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  5. Isidoro López-Braña
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Correspondence to Isidoro López-Braña.

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Communicated by D. Zaitlin.

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Simonetti, E., Alba, E., Montes, M.J. et al. Analysis of ascorbate peroxidase genes expressed in resistant and susceptible wheat lines infected by the cereal cyst nematode, Heterodera avenae . Plant Cell Rep 29, 1169–1178 (2010). https://doi.org/10.1007/s00299-010-0903-z

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