A class III peroxidase PRX34 is a component of disease resistance in Arabidopsis

Abstract

PRX34 mediates the oxidative burst in Arabidopsis. Here we characterized two additional Arabidopsis prx34 null mutants (prx34-2, prx34-3), besides the well-studied prx34-1. Due to a decrease in corresponding peroxidase, the activity that generates reactive oxygen species (ROS) was significantly lower in cell wall extracts of prx34-2 and prx34-3 plants. Consistently, the prx34-2 and prx34-3 exhibited reduced accumulation both of ROS and callose in Flg22-elicitor-treated leaves, leading to enhanced susceptibility to bacterial and fungal pathogens. In contrast, ectopic expression of PRX34 in the wild type caused enhanced resistance. PRX34 is thus a component for disease resistance in Arabidopsis.

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References

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

    CAS  Article  Google Scholar 

  2. Alonso JM, Stepanova AN, Leisse TJ, Kim CJ, Chen H, Shinn P, Stevenson DK, Zimmerman J, Barajas P, Cheuk R, Gadrinab C, Heller C, Jeske A, Koesema E, Meyers CC, Parker H, Prednis L, Ansari Y, Choy N, Deen H, Geralt M, Hazari N, Hom E, Karnes M, Mulholland C, Ndubaku R, Schmidt I, Guzman P, Aguilar-Henonin L, Schmid M, Weigel D, Carter DE, Marchand T, Risseeuw E, Brogden D, Zeko A, Crosby WL, Berry CC, Ecker JR (2003) Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science 301:653–657

    Article  Google Scholar 

  3. Arnaud D, Lee S, Takebayashi Y, Choi D, Choi J, Sakakibara H, Hwang I (2017) Cytokinin-mediated regulation of reactive oxygen species homeostasis modulates stomatal immunity in Arabidopsis. Plant Cell 29:543–559

    CAS  Article  Google Scholar 

  4. Bestwick CS, Brown IR, Mansfield JW (1998) Localized changes in peroxidase activity accompany hydrogen peroxide generation during the development of a nonhost hypersensitive reaction in lettuce. Plant Physiol 118:1067–1078

    CAS  Article  Google Scholar 

  5. 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

    CAS  Article  Google Scholar 

  6. Bolwell GP, Davies DR, Gerrish C, Auh CK, Murphy TM (1998) Comparative biochemistry of the oxidative burst produced by rose and French bean cells reveals two distinct mechanisms. Plant Physiol 116:1379–1385

    CAS  Article  Google Scholar 

  7. 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

    CAS  PubMed  Google Scholar 

  8. Choi HW, Kim YJ, Lee SC, Hong KJ, Hwang BK (2007) Hydrogen peroxide generation by the pepper extracellular peroxidase CaPO2 activates local and systemic cell death and defense response to bacterial pathogens. Plant Physiol 145:890–904

    CAS  Article  Google Scholar 

  9. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743

    CAS  Article  Google Scholar 

  10. Daudi A, Cheng Z, O’Brien JA, Mammarella N, Khan S, Ausubel FM, Bolwell GP (2012) The apoplastic oxidative burst peroxidase in Arabidopsis is a major component of pattern-triggered immunity. Plant Cell 24:275–287

    CAS  Article  Google Scholar 

  11. Davies DR, Bindschedler LV, Strickland TS, Bolwell GP (2006) Production of reactive oxygen species in Arabidopsis thaliana cell suspension cultures in response to an elicitor from Fusarium oxysporum: implications for basal resistance. J Exp Bot 57:1817–1827

    CAS  Article  Google Scholar 

  12. Gross GG, Janse C, Elstner EF (1977) Involvement of malate, monophenols, and the superoxide radical in hydrogen peroxide formation by isolated cell walls from horseradish (Armoracia lapathifolia Gilib.). Planta 136:271–276

    CAS  Article  Google Scholar 

  13. Halliwell B (1978) Lignin synthesis: the generation of hydrogen peroxide and superoxide by horseradish peroxidase and its stimulation by manganese (II) and phenols. Planta 140:81–88

    CAS  Article  Google Scholar 

  14. 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 

  15. Kiba A, Miyake C, Toyoda K, Ichinose Y, Yamada T, Shiraishi T (1997) Superoxide generation in extracts from isolated plant cell walls is regulated by fungal signal molecules. Phytopathology 87:846–852

    CAS  Article  Google Scholar 

  16. Kimura M, Kawano T (2015) Hydrogen peroxide-independent generation of superoxide catalyzed by soybean peroxidase in response to ferrous ion. Plant Signal Behav 10:e1010917. https://doi.org/10.1080/15592324.2015.1010917

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  17. Lyons R, Stiller J, Powell J, Rusu A, Manners JM, Kazan K (2015) Fusarium oxysporum triggers tissue-specific transcriptional reprogramming in Arabidopsis thaliana. PLoS One 10:e0121902

    Article  Google Scholar 

  18. Mammarella ND, Cheng Z, Fu ZQ, Daudi A, Bolwell GP, Dong X, Ausubel FM (2015) Apoplastic peroxidases are required for salicylic acid-mediated defense against Pseudomonas syringae. Phytochemisry 112:110–121

    CAS  Article  Google Scholar 

  19. Matsui H, Nomura Y, Egusa M, Hamada T, Hyon GS, Kaminaka H, Watanabe Y, Ueda T, Trujillo M, Shirasu K, Nakagami H (2017) The GYF domain protein PSIG1 dampens the induction of cell death during plant–pathogen interactions. PLoS Genet 13:e1007037

    Article  Google Scholar 

  20. Nakagawa T, Kurose T, Hino T, Tanaka K, Kawamukai M, Niwa Y, Toyooka K, Matsuoka K, Jindo T, Kimura T (2007) Development of series of gateway binary vectors, pGWBs, for realizing efficient construction of fusion genes for plant transformation. J Biosci Bioeng 104:34–41

    CAS  Article  Google Scholar 

  21. O’Brien JA, Daudi A, Butt VS, Bolwell GP (2012a) Reactive oxygen species and their role in plant defence and cell wall metabolism. Planta 236:765–779

    Article  Google Scholar 

  22. O’Brien JA, Daudi A, Finch P, Butt VS, Whitelegge JP, Souda P, Ausubel FM, Bolwell GP (2012b) A peroxidase-dependent apoplastic oxidative burst in cultured Arabidopsis cells functions in MAMP-elicited defense. Plant Physiol 158:2013–2027

    Article  Google Scholar 

  23. Oliva M, Theiler G, Zamocky M, Koua D, Margis-Pinheiro M, Passardi F, Dunand C (2009) PeroxiBase: a powerful tool to collect and analyse peroxidase sequences from Viridiplantae. J Exp Bot 60:453–459

    CAS  Article  Google Scholar 

  24. Passardi F, Tognolli M, DeMeyer M, Penel C, Dunand C (2006) Two cell wall associated peroxidases from Arabidopsis influence root elongation. Planta 223:965–974

    CAS  Article  Google Scholar 

  25. Shigeto J, Tsutsumi Y (2016) Diverse functions and reactions of class III peroxidases. New Phytol 209:1395–1402

    CAS  Article  Google Scholar 

  26. Tognolli M, Penel C, Greppin H, Simon P (2002) Analysis and expression of the class III peroxidase large gene family in Arabidopsis thaliana. Gene 288:129–138

    CAS  Article  Google Scholar 

  27. Toyoda K, Yasunaga E, Niwa M, Ohwatari Y, Nakashima A, Inagaki Y, Ichinose Y, Shiraishi T (2012) H2O2 production by copper amine oxidase, a component of the ecto-apyrase (ATPase)-containing protein complex(es) in the pea cell wall, is regulated by an elicitor and a suppressor from Mycosphaerella pinodes. J Gen Plant Pathol 78:311–315

    CAS  Article  Google Scholar 

  28. Welinder KG, Justesen AF, Kjærsgård IVH, Jensen RB, Rasmussen SK, Jespersen HM, Duroux L (2002) Structural diversity and transcription of class III peroxidases from Arabidopsis thaliana. FEBS J 269:6063–6081

    CAS  Google Scholar 

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Acknowledgements

We acknowledge Prof. Dr. Shinji Tsuyumu (Laboratory of Plant Pathology, Faculty of Agriculture, Shizuoka University, Shizuoka, Japan) for generously providing Pectobacterium carotovorum subsp. carotovorum strain Pc1. This research was supported in part by the Grants-in-Aid for Scientific Research (18K05645) from the Japan Society for Promotion of Science (JSPS).

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Correspondence to Kazuhiro Toyoda.

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Fig. S1 Response of prx34-1 mutants to fungal and bacterial pathogens. a Symptoms and area of lesions induced by Botrytis cinerea (MAFF712189), Colletotrichum higginsianum (MAFF305635) and Pectobacterium carotovorum subsp. carotovorum strain Pc1. Inoculations: B. cinerea and C. higginsianum, 5-μl drop of 2 × 105 conidia/ml in distilled water or 1% Sabouraud maltose broth was placed on each side of the mid vein of detached leaves of 4-week-old wild type or prx34-1; P. carotovorum subsp. carotovorum strain Pc1, 5-μl drop of 1×105 cfu/ml was placed on wounded leaves. All inoculated leaves were incubated for 2 or 3 days at 22 °C, then photographed, and lesions were measured. Data are the average ± SD of 10 leaves from five independent plants. No significant difference was observed between the wild type and mutant prx34-1, as revealed by Dunnett’s test. b Flg22-elicted ROS generation in Arabidopsis leaves. Mature leaves of 4-week-old seedlings of the wild type or prx34-1 were infiltrated with approximately 0.1 ml of 100 nM Flg22 or water (as control). The leaves were stained with 3,3′-diaminobenzidine (DAB) at 2 h after treatment to detect H2O2. Experiments were repeated three times with similar results. c Flg22-elicited callose detected by aniline blue staining. Mature leaves were infiltrated with approximately 0.1 ml of 100 nM Flg22 or water (as control) as described above. Leaves were stained with aniline blue 24 h after treatment to detect callose. The number of callose deposits was calculated using ImageJ software. Data are the average ± SD of 10 leaves from five independent plants. Asterisks indicate significant difference (Dunnett’s test; ***p < 0.001).

Fig. S2 Genotyping of three prx34 mutants. PCR was performed with a Phire Plant Direct PCR Kit (Thermo Fisher Scientific) using genomic DNAs of mutant prx34-1 (SALK_051769) (a), prx34-2 (GABI_728F08) (b) or prx34-3 (SALK_112466C) (c). Control amplification was carried out with primers supplied with the kit that amplifies a 297-bp fragment of a highly conserved region of chloroplast DNA. Wild type Col-0 was used as the control. The T-DNA border primer (BP), left (LP) and right (RP) genomic primers for each mutant line were designed using T-DNA Primer Design (http://signal.salk.edu/tdnaprimers.2.html) and are listed in Table S1. The RP was always on the side of the flanking sequence (the 3′ end of the insertion). Therefore, by using the three primers (BP+LP+RP), the wild type (no insertion) yielded an amplicon of about 1100 bp (from LP to RP). For homozygous insertion lines, the PCR amplified a distinct band of 410 + N bp (from RP to insertion site 300 + N bases, plus 110 bases from BP to the left border of the vector).

Fig. S3 Induction of PRX34 transcripts in Arabidopsis in response to Flg22 or chitoheptaose. Mature leaves of 4-week-old wild type seedlings were infiltrated with approximately 0.1 ml of 100 nM Flg22 (GenScript, Piscataway, NJ, USA), 100 μg/ml chitoheptaose (GLU437; Elicityl, Crolles, France) or water (as control). Leaves were harvested 6, 12 and 24 h after infiltration, then subjected to quantitative RT-PCR using a Shimadzu GVP-9600 Gene Detection System (Shimadzu, Kyoto, Japan) and primers in Supplementary Table 2. Expression level of PRX34 was normalized using EF1-α (At5g60390) and expressed relative to the water-treated control sample. Data are the average ± standard deviation (SD) from three independent plants. Asterisks indicate statistically significant difference (Dunnett’s test; *p < 0.05; ***p < 0.001).

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Zhao, L., Phuong, L.T., Luan, M.T. et al. A class III peroxidase PRX34 is a component of disease resistance in Arabidopsis. J Gen Plant Pathol 85, 405–412 (2019). https://doi.org/10.1007/s10327-019-00863-9

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Keywords

  • Apoplastic oxidative burst
  • Arabidopsis
  • Cell wall
  • Class III peroxidase
  • PRX34
  • Reactive oxygen species (ROS)