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Dynamics and localization of H2O2 production in elicited plant cells

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Summary.

H2O2 produced in plant cells plays a dual role. In addition to its antimicrobial effect, it also acts as a secondary messenger initiating and modulating responses of plants exposed to unfavorable external signals. A suspension culture of Rubia tinctorum cells challenged with elicitors was used as a model system to investigate H2O2 formation. Cellular H2O2 was measured by a modified titanium(IV) method, while that in the medium was detected with scopoletin fluorescence. Localization of H2O2 production at the ultrastructural level was carried out by the CeCl3 reaction. A fungal elicitor induced H2O2 production with transient maxima, the first of which appeared 4 min after treatment. Three subsequent maxima appeared in the cells up to 48 h after treatment. Exposure of cells to exogenous jasmonic acid and salicylic acid also changed the H2O2 concentration maxima over 48 h; however, their timing was slightly shifted. Fungal-elicitor, jasmonic acid, and salicylic acid treatments had different effects on the H2O2 concentration in the medium. Ultrastructural investigations revealed that electron-dense precipitates were present at the plasmalemma and in some nearby vesicular cytoplasmic structures 30 min after treatment. Later samples showed cytochemical-precipitate accumulation in the cell walls. These deposits appeared to be local and independent of the direction of the external signal. We could not detect the presence of H2O2 in peroxisomes, mitochondria, plastids, or the central vacuolar space. Electron energy loss spectroscopy investigations distinguished between the cerium-containing precipitates and other electrondense particles, thereby proving that H2O2 generation occurs locally.

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Abbreviations

JA:

jasmonic acid

SA:

salicylic acid

EELS:

electron energy loss spectroscopy

ROS:

reactive oxygen species

SEM:

scanning electron microscope

TEM:

transmission electron microscope

PEELS:

parallel electron energy loss spectrum

References

  • A Ádám T Farkas G Somlyai M Hevesi Z Király (1989) ArticleTitleConsequence of O2 •-generation during a bacterially induced hypersensitive reaction in tobacco: deterioration of membrane lipids Physiol Mol Plant Pathol 34 13–26 Occurrence Handle10.1016/0885-5765(89)90013-1

    Article  Google Scholar 

  • S Agarwal RK Sairam GC Srivastava RC Meena (2005) ArticleTitleChanges in antioxidant enzymes activity and oxidative stress by abscisic acid and salicylic acid in wheat genotypes Biol Plant 49 541–550 Occurrence Handle10.1007/s10535-005-0048-z Occurrence Handle1:CAS:528:DC%2BD28XlvFeiuw%3D%3D

    Article  CAS  Google Scholar 

  • CC Ahn OL Krivanek (1983) EELS atlas: a reference guide of electron energy loss spectra covering all stable elements Gatan and Arizona State University HREM Facility Warrendale, Pa

    Google Scholar 

  • K Apel H Hirt (2004) ArticleTitleReactive oxygen species: metabolism, oxidative stress, and signal transduction Annu Rev Plant Biol 55 373–399 Occurrence Handle15377225 Occurrence Handle10.1146/annurev.arplant.55.031903.141701 Occurrence Handle1:CAS:528:DC%2BD2cXlvFeisL0%3D

    Article  PubMed  CAS  Google Scholar 

  • CJ Baker EW Orlandi (1995) ArticleTitleActive oxygen in plant pathogenesis Annu Rev Phytopathol 33 299–321 Occurrence Handle10.1146/annurev.py.33.090195.001503 Occurrence Handle1:CAS:528:DyaK2MXosFWisLg%3D Occurrence Handle18999963

    Article  CAS  PubMed  Google Scholar 

  • CJ Baker NR O’Neill LD Keppler EW Orlandi (1991) ArticleTitleEarly responses during plant–bacteria interactions in tobacco cell suspensions Phytopathology 81 1504–1507

    Google Scholar 

  • CS Bestwick IR Brown MHR Bennett JW Mansfield (1997) ArticleTitleLocalization of hydrogen peroxide accumulation during the hypersensitive reaction of lettuce cells to Pseudomonas syringae pv phaseolicola Plant Cell 9 209–221 Occurrence Handle9061952 Occurrence Handle10.1105/tpc.9.2.209 Occurrence Handle1:CAS:528:DyaK2sXhslGktb4%3D

    Article  PubMed  CAS  Google Scholar 

  • K Bóka J Jakab I Király (2002) ArticleTitleComparison of the effect of different fungal elicitors on Rubia tinctorum L. suspension culture Biol Plant 45 281–290 Occurrence Handle10.1023/A:1015113226897

    Article  Google Scholar 

  • T Brennan C Frenkel (1977) ArticleTitleInvolvement of hydrogen peroxide in the regulation of senescence in pear Plant Physiol 59 411–416 Occurrence Handle16659863 Occurrence Handle1:CAS:528:DyaE2sXhs1yjsL4%3D Occurrence Handle10.1104/pp.59.3.411

    Article  PubMed  CAS  Google Scholar 

  • AC Cazalé MA Rouet-Mayer H Barbier-Brygoo Y Mathieu C Lauriere (1998) ArticleTitleOxidative burst and hypoosmotic stress in tobacco cell suspensions Plant Physiol 116 659–669 Occurrence Handle9490766 Occurrence Handle10.1104/pp.116.2.659

    Article  PubMed  Google Scholar 

  • Z Chen H Silva DF Klessig (1993) ArticleTitleActive oxygen species in the induction of plant systemic acquired resistance by salicylic acid Science 262 1883–1886 Occurrence Handle8266079 Occurrence Handle10.1126/science.8266079 Occurrence Handle1:CAS:528:DyaK2cXhtVWqu7o%3D

    Article  PubMed  CAS  Google Scholar 

  • RA Creelman JE Mullet (1997) ArticleTitleOligosaccharins, brassinolides, and jasmonates: nontraditional regulators of plant growth, development, and gene expression Plant Cell 9 1211–1223 Occurrence Handle9254935 Occurrence Handle10.1105/tpc.9.7.1211 Occurrence Handle1:CAS:528:DyaK2sXlt1ShtbY%3D

    Article  PubMed  CAS  Google Scholar 

  • Y Czaninski RM Sachot AM Catesson (1993) ArticleTitleCytochemical localization of hydrogen peroxide in lignifying cell walls Ann Bot 72 547–550 Occurrence Handle10.1006/anbo.1993.1143 Occurrence Handle1:CAS:528:DyaK2cXisVShsL0%3D

    Article  CAS  Google Scholar 

  • R Desikan JT Hancock MJ Coffey (1996) ArticleTitleGeneration of active oxygen in elicited cells of Arabidopsis thaliana is mediated by NADPH oxidase-like enzyme FEBS Lett 382 213–217 Occurrence Handle8612756 Occurrence Handle10.1016/0014-5793(96)00177-9 Occurrence Handle1:CAS:528:DyaK28XhslehtrY%3D

    Article  PubMed  CAS  Google Scholar 

  • M Fauth A Merten MG Hahn W Jeblick H Kauss (1996) ArticleTitleCompetence for elicitation of H2O2 in hypocotyls of cucumber is induced by breaching the cuticle and is enhanced by salicylic acid Plant Physiol 110 347–354 Occurrence Handle12226186 Occurrence Handle1:CAS:528:DyaK28XhtF2nur0%3D

    PubMed  CAS  Google Scholar 

  • J Fodor G Gullner AL Ádám B Barna T Kömives Z Király (1997) ArticleTitleLocal and systemic responses of antioxidants to tobacco mosaic virus and to salicylic acid in tobacco Plant Physiol 114 1443–1451 Occurrence Handle12223782 Occurrence Handle1:CAS:528:DyaK2sXlsleitrs%3D

    PubMed  CAS  Google Scholar 

  • V Ganesan G Thomas (2001) ArticleTitleSalicylic acid response in rice: influence of salicylic acid on H2O2 accumulation and oxidative stress Plant Sci 160 1095–1106 Occurrence Handle11337066 Occurrence Handle10.1016/S0168-9452(01)00327-2 Occurrence Handle1:CAS:528:DC%2BD3MXjtFGjur4%3D

    Article  PubMed  CAS  Google Scholar 

  • LV Gómez Ros A Paradiso C Gabaldón MA Pedreño L de Gara A Ros Barceló (2006) ArticleTitleTwo distinct cell sources of H2O2 in the lignifying Zinnia elegans cell culture system Protoplasma 227 175–183 Occurrence Handle16520879 Occurrence Handle10.1007/s00709-005-0128-0 Occurrence Handle1:CAS:528:DC%2BD28XltFWksL8%3D

    Article  PubMed  CAS  Google Scholar 

  • R Gupta S Luan (2003) ArticleTitleRedox control of protein tyrosine phosphatases and mitogen-activated protein kinases in plants Plant Physiol 132 1149–1152 Occurrence Handle12857797 Occurrence Handle10.1104/pp.103.020792 Occurrence Handle1:CAS:528:DC%2BD3sXlsFGht70%3D

    Article  PubMed  CAS  Google Scholar 

  • I Hwang H-C Chen J Sheen (2002) ArticleTitleTwo component signal transduction pathways in Arabidopsis Plant Physiol 129 500–515 Occurrence Handle12068096 Occurrence Handle10.1104/pp.005504 Occurrence Handle1:CAS:528:DC%2BD38XkvV2jsb0%3D

    Article  PubMed  CAS  Google Scholar 

  • M Iwano F-S Che K Goto N Tanaka S Takayama A Isogai (2002) ArticleTitleElectron microscopic analysis of the H2O2 accumulation preceding hypersensitive cell death induced by an incompatible strain of Pseudomonas avenae in cultured rice cells Mol Plant Pathol 3 1–8 Occurrence Handle10.1046/j.1464-6722.2001.00087.x Occurrence Handle1:CAS:528:DC%2BD38XitVKmtr4%3D

    Article  CAS  Google Scholar 

  • C Jonak L Ökresz L Bögre H Hirt (2002) ArticleTitleComplexity, cross talk and integration of plant MAP kinase signaling Curr Opin Plant Biol 5 415–424 Occurrence Handle12183180 Occurrence Handle10.1016/S1369-5266(02)00285-6 Occurrence Handle1:CAS:528:DC%2BD38XmtlGjtLY%3D

    Article  PubMed  CAS  Google Scholar 

  • H Kauss W Jeblick (1995) ArticleTitlePretreatment of parsley suspension cultures with salicylic acid enhances spontaneous and elicited production of H2O2 Plant Physiol 108 1171–1178 Occurrence Handle12228535 Occurrence Handle1:CAS:528:DyaK2MXmvFGmur0%3D

    PubMed  CAS  Google Scholar 

  • H Kauss W Jeblick (1996) ArticleTitleInfluence of salicylic acid on the induction of competence for H2O2 elicitation Plant Physiol 111 755–763 Occurrence Handle12226328 Occurrence Handle1:CAS:528:DyaK28XktlKks7w%3D

    PubMed  CAS  Google Scholar 

  • T Kawano (2003) ArticleTitleRoles of the reactive oxygen species-generating peroxidase reactions in plant defense and growth induction Plant Cell Rep 21 829–837 Occurrence Handle12789499 Occurrence Handle1:CAS:528:DC%2BD3sXjvVSlt74%3D

    PubMed  CAS  Google Scholar 

  • I Kottke T Holopainen E Alanen K Turnau (1995) ArticleTitleDeposition of nitrogen in vacuolar bodies of Cenococcum geophilum Fr. mycorrhizas as detected by electron energy loss spectroscopy New Phytol 126 411–416 Occurrence Handle10.1111/j.1469-8137.1995.tb04311.x

    Article  Google Scholar 

  • Z Kristóf O Tímár K Imre (1999) ArticleTitleChanges of calcium distribution in ovules of Torenia fournieri during pollination and fertilization Protoplasma 208 149–155 Occurrence Handle10.1007/BF01279085

    Article  Google Scholar 

  • C Lamb RA Dixon (1997) ArticleTitleThe oxidative burst in plant disease resistance Annu Rev Plant Physiol Plant Mol Biol 48 251–275 Occurrence Handle15012264 Occurrence Handle10.1146/annurev.arplant.48.1.251 Occurrence Handle1:CAS:528:DyaK2sXjs1entr8%3D

    Article  PubMed  CAS  Google Scholar 

  • L Legendre S Reuter FP Heinstein SP Low (1993) ArticleTitleCharacterization of the oligogalacturonide-induced oxidative burst in cultured soybean (Glycine max) cells Plant Physiol 102 233–240 Occurrence Handle12231814 Occurrence Handle1:CAS:528:DyaK3sXks1yhs78%3D

    PubMed  CAS  Google Scholar 

  • L Liu KEL Eriksson JFD Dean (1999) ArticleTitleLocalization of hydrogen peroxide production in Zinnia elegans L. stems Phytochemistry 52 545–554 Occurrence Handle10.1016/S0031-9422(99)00231-9 Occurrence Handle1:CAS:528:DyaK1MXntlOjs7s%3D

    Article  CAS  Google Scholar 

  • PS Low JR Merida (1996) ArticleTitleThe oxidative burst in plant defense: function and signal transduction Physiol Plant 96 533–542 Occurrence Handle10.1111/j.1399-3054.1996.tb00469.x Occurrence Handle1:CAS:528:DyaK28XivV2qtLc%3D

    Article  CAS  Google Scholar 

  • U Lütz-Meindl C Lütz (2006) ArticleTitleAnalysis of element accumulation in cell wall attached and intracellular particles of snow algae by EELS and ESI Micron 37 452–458 Occurrence Handle16376553 Occurrence Handle10.1016/j.micron.2005.11.004 Occurrence Handle1:CAS:528:DC%2BD28XmtlOhtbg%3D

    Article  PubMed  CAS  Google Scholar 

  • MC Mehdy YK Sharma K Sathasivan NW Bays (1996) ArticleTitleThe role of activated oxygen species in plant disease resistance Physiol Plant 98 365–374 Occurrence Handle10.1034/j.1399-3054.1996.980219.x Occurrence Handle1:CAS:528:DyaK28XmsVCntbs%3D

    Article  CAS  Google Scholar 

  • DG Mellersh IV Foulds J Verna HC Heath MC Heath (2002) ArticleTitleH2O2 plays different roles in determining penetration failure in three diverse plant-fungal interactions Plant J 29 257–268 Occurrence Handle11844104 Occurrence Handle10.1046/j.0960-7412.2001.01215.x Occurrence Handle1:CAS:528:DC%2BD38XhvFOit7k%3D

    Article  PubMed  CAS  Google Scholar 

  • T Murashige F Skoog (1962) ArticleTitleRevised medium for rapid growth and bioassays with tobacco tissue cultures Physiol Plant 15 473–497 Occurrence Handle10.1111/j.1399-3054.1962.tb08052.x Occurrence Handle1:CAS:528:DyaF3sXksFKm

    Article  CAS  Google Scholar 

  • T Niki I Mitsuhara S Seo N Ohtsubo Y Ohashi (1998) ArticleTitleAntagonistic effect of salicylic acid and jasmonic acid on the expression of pathogenesis-related (PR) protein genes in wounded mature tobacco leaves Plant Cell Physiol 39 500–507 Occurrence Handle1:CAS:528:DyaK1cXjsFSmt78%3D

    CAS  Google Scholar 

  • E Olmos JR Martínez-Solano A Piqueras E Hellín (2003) ArticleTitleEarly steps in the oxidative burst induced by cadmium in cultured tobacco cells J Exp Bot 54 291–301 Occurrence Handle12493856 Occurrence Handle10.1093/jxb/54.381.291 Occurrence Handle1:CAS:528:DC%2BD3sXhtlOiu7w%3D

    Article  PubMed  CAS  Google Scholar 

  • PD Olson JE Varner (1993) ArticleTitleHydrogen peroxide and lignification Plant J 4 887–892 Occurrence Handle10.1046/j.1365-313X.1993.04050887.x Occurrence Handle1:CAS:528:DyaK2cXitFWktrg%3D

    Article  CAS  Google Scholar 

  • ML Orozco-Cárdenas J Narváez-Vásquez CA Ryan (2001) ArticleTitleHydrogen peroxide acts as a second messenger for the induction of defense genes in tomato plants in response to wounding, systemin, and methyl jasmonate Plant Cell 13 179–191 Occurrence Handle11158538 Occurrence Handle10.1105/tpc.13.1.179

    Article  PubMed  Google Scholar 

  • JC Promé (1996) ArticleTitleSignalling events elicited in plants by defined oligosaccharide structures Curr Opin Struct Biol 6 671–678 Occurrence Handle8913691 Occurrence Handle10.1016/S0959-440X(96)80035-2

    Article  PubMed  Google Scholar 

  • MV Rao G Paliyath DP Ormrod DP Murr CB Watkins (1997) ArticleTitleInfluence of salicylic acid on H2O2 production, oxidative stress and H2O2-metabolizing enzymes Plant Physiol 115 137–149 Occurrence Handle9306697 Occurrence Handle10.1104/pp.115.1.137 Occurrence Handle1:CAS:528:DyaK2sXmt1yntLc%3D

    Article  PubMed  CAS  Google Scholar 

  • V Repka (2006) ArticleTitleEarly defence responses induced by two distinct elicitors derived from a Botrytis cinerea in grapevine leaves and cell suspensions Biol Plant 50 94–106 Occurrence Handle10.1007/s10535-005-0080-z Occurrence Handle1:CAS:528:DC%2BD28Xjtlertbk%3D

    Article  CAS  Google Scholar 

  • ES Reynolds (1963) ArticleTitleThe use of lead citrate at high pH as an electron-opaque stain in electron microscopy J Cell Biol 17 208–212 Occurrence Handle13986422 Occurrence Handle10.1083/jcb.17.1.208 Occurrence Handle1:CAS:528:DyaF3sXktVClu70%3D

    Article  PubMed  CAS  Google Scholar 

  • M Rojkind J-A Domínguez-Rosales N Nieto P Greenwel (2002) ArticleTitleRole of hydrogen peroxide and oxidative stress in healing responses Cell Mol Life Sci 59 1872–1891 Occurrence Handle12530519 Occurrence Handle10.1007/PL00012511 Occurrence Handle1:CAS:528:DC%2BD38Xps1entbs%3D

    Article  PubMed  CAS  Google Scholar 

  • RS Ros-Barceló F Pomar M López-Serrano P Martínez MA Pedreño (2002) ArticleTitleDevelopmental regulation of the H2O2-producing system and of a basic peroxidase isoenzyme in the Zinnia elegans lignifying xylem Plant Physiol Biochem 40 325–332 Occurrence Handle10.1016/S0981-9428(02)01376-1

    Article  Google Scholar 

  • H Thordal-Christensen Z Zhang Y Wei DB Collinge (1997) ArticleTitleSubcellular localization of H2O2 in plants: H2O2 accumulation in papillae and hypersensitive response during the barley–powdery mildew interaction Plant J 11 1187–1194 Occurrence Handle10.1046/j.1365-313X.1997.11061187.x Occurrence Handle1:CAS:528:DyaK2sXkslajtLs%3D

    Article  CAS  Google Scholar 

  • A Vasconsuelo AM Giulietti G Picotto J Rodriguez-Talou R Boland (2003) ArticleTitleInvolvement of the PLC/PKC pathway in chitosan-induced anthraquinone production by Rubia tinctorum L. cell cultures Plant Sci 163 429–436 Occurrence Handle10.1016/S0168-9452(03)00208-5 Occurrence Handle1:CAS:528:DC%2BD3sXkvF2jurc%3D

    Article  CAS  Google Scholar 

  • A Vasconsuelo AM Giulietti R Boland (2004) ArticleTitleSignal transduction events mediating chitosan stimulation of anthraquinone synthesis in Rubia tinctorum Plant Sci 166 405–413 Occurrence Handle10.1016/j.plantsci.2003.10.007 Occurrence Handle1:CAS:528:DC%2BD2cXntVOrtQ%3D%3D

    Article  CAS  Google Scholar 

  • Y Yang M Qi C Mei (2004) ArticleTitleEndogenous salicylic acid protects rice plants from oxidative damage caused by aging as well as biotic and abiotic stress Plant J 40 909–919 Occurrence Handle15584956 Occurrence Handle10.1111/j.1365-313X.2004.02267.x Occurrence Handle1:CAS:528:DC%2BD2MXpslGhsQ%3D%3D

    Article  PubMed  CAS  Google Scholar 

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  1. Department of Plant Anatomy, Eötvös University, Budapest

    K. Bóka, N. Orbán & Z. Kristóf

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  1. K. Bóka
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  2. N. Orbán
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  3. Z. Kristóf
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Correspondence to K. Bóka.

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Correspondence and reprints: Department of Plant Anatomy, Eötvös University, Pázmány Péter sétány 1/C, Budapest, 1117 Hungary.

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Bóka, K., Orbán, N. & Kristóf, Z. Dynamics and localization of H2O2 production in elicited plant cells. Protoplasma 230, 89–97 (2007). https://doi.org/10.1007/s00709-006-0225-8

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