Abstract
Previous studies have shown that hydrogen peroxide (H2O2) may mediate the auxin response during the formation of adventitious roots (AR). However, the mechanism and distribution of H2O2 during AR formation remains unclear. In this study, we investigate the spatiotemporal changes and role of H2O2 in AR initiation and development. Application of 5–100 mM H2O2 to Mung bean (Phaseolus radiatus L.) hypocotyl cuttings induced AR formation in a dose-dependent manner. The effect was blocked by ascorbic acid (AA), an important reducing substrate for H2O2 reduction. Depletion of endogenous H2O2 by AA resulted in the significant reduction of AR emergence, suggesting a physiological role for H2O2 in the regulation of AR formation. Determination of H2O2 content showed that the level of H2O2 increased gradually and reached the highest value 60 h after induction of AR. Further detection of endogenous H2O2 by the specific fluorescent probe dichlorofluorescein diacetate (H2DCF-DA) and 3,3′-diaminobenzidine (DAB) staining in transverse sections of the basal region of cuttings revealed that obvious H2O2 signals were observed in the pericycle cells between the vascular bundles 24 h after the primary roots were removed. With the development of root primordia, H2O2 signals increased gradually and were mainly distributed in the root meristem. AA significant inhibited the H2O2-dependent fluorescence and the formation of AR, suggesting an essential role of H2O2 generation during AR initiation and development. Furthermore, the involvement of Ca2+ during H2O2-mediated AR formation was evaluated. Ca2+ channel inhibitors LaCl3 and ruthenium red (RR) and Ca2+ chelator ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA) prevent H2O2-induced AR formation, which indicate that the hypocotyl cuttings response to H2O2 depends on the availability of both intracellular and extracellular Ca2+ pools, and Ca2+ is a downstream messenger in the signaling pathway triggered by H2O2 to promote adventitious root formation.
Similar content being viewed by others
References
Allan AC, Fluhr R (1997) Two distinct sources of elicited reactive oxygen species in tobacco epidermal cells. Plant Cell 9:1559–1572
Allen GJ, Muir SR, Sanders D (1995) Release of Ca2+ from individual plant vacuoles by both InsP3 and cyclic ADP-ribose. Science 268:735–737
Barceló AR, Pomar F, López-Serrano M, Pedreño MA (2003) Peroxidase: a multifunctional enzyme in grapevines. Funct Plant Biol 30:577–591
Beers EP, McDowell JM (2001) Regulation and execution of programmed cell death in response to pathogens, stress and developmental cues. Curr Opin Plant Biol 4:561–567
Bestwick CS, Brown IR, Bennett MHR, Mansfield JW (1997) Localization of hydrogen peroxide accumulation during the hypersensitive reaction of lettuce cells to Pseudomonas syringae pv phaseolicola. Plant Cell 9:209–221
Bolwell GP (1999) Role of active oxygen species and NO in plant defense responses. Curr Opin Plant Biol 2:287–294
Bolwell GP, Butt VS, Davies DR, Zimmerlin A (1995) The origin of the oxidative burst in plants. Free Radic Res 23:517–532
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
Boo YC, Jung J (1999) Water deficit: induced oxidative stress and antioxidative defenses in rice plants. J Plant Physiol 155:255–261
Brennan T, Frenkel C (1977) Involvement of hydrogen peroxide in the regulation of senescence in pear. Plant Physiol 59:411–416
Bush DS (1995) Calcium regulation in plants and its role in signaling. Ann Rev Plant Physiol Plant Mol Biol 46:95–112
Clayton H, Knight MR, Knight H, McAinsh MR, Hetherington AM (1999) Dissection of the ozone-induced calcium signature. Plant J 17:575–579
de Marco A, Roubelakis-Angelakis KA (1996) Hydrogen peroxide plays a bivalent role in the regeneration of protoplasts. J Plant Physiol 149:109–114
Desikan R, Cheung M-K, Bright J, Henson D, Hancock JT, Neill SJ (2004) ABA, hydrogen peroxide and nitric oxide signalling in stomatal guard cells. J Exp Bot 55:205–212
Evans NH, McAinsh MR, Hetherington AM (2001) Calcium oscillations in higher plants. Curr Opin Plant Biol 4:415–420
Foreman J, Demidchik V, Bothwell JHF, Mylona P, Miedema H, Torres MA, Linstead P, Costa S, Brownlee C, Jones JDG (2003) Reactive oxygen species produced by NADPH oxidase regulate plant cell growth. Nature 42:442–446
Gaspar T, Kevers C, Hausman JF (1997) Indissociable chief factors in the inductive phase of adventitious rooting. In: Altman A, Waisel Y (eds) Biology root formation and development. Plenum Press, New York, pp 55–63
Huang AX, She XP, Huang C, Song TS (2007) The dynamic distribution of NO and NADPH-diaphorase activity during IBA-induced adventitious root formation. Physiol Plant 130:240–249
Jiang M, Zhang J (2002) Involvement of plasma-membrane NADPH oxidase in abscisic acid- and water stress-induced antioxidant defense in leaves of maize seedlings. Planta 215:1022–1030
Joo JH, Bae YS, Lee JS (2001) Role of auxin-induced reactive oxygen species in root gravitropism. Plant Physiol 126:1055–1060
Kwak JM, Nguyen V, Schroeder JI (2006) The role of reactive oxygen species in hormonal responses. Plant Physiol 141:323–329
Lamb C, Dixon RA (1997) The oxidative burst in plant disease resistance. Ann Rev Plant Physiol Plant Mol Biol 48:251–275
Lanteri ML, Pagnussat GC, Lamattina L (2006) Calcium and calcium-dependent protein kinases are involved in nitric oxide- and auxin-induced adventitious root formation in cucumber. J Exp Bot 57:1341–1351
Lanteri ML, Laxalt AM, Lamattina L (2008) Nitric oxide triggers phosphatidic acid accumulation via phospholipase D during auxin-induced adventitious root formation in cucumber. Plant Physiol 147:188–198
Li S, Xue L, Xu S, Feng H, An L (2007) Hydrogen peroxide involvement in formation and development of adventitious roots in cucumber. Plant Growth Regul 52:173–180
Mackerness SAH, John CF, Jordan B, Thomas B (2001) Early signaling components in ultraviolet-B responses: distinct roles for different reactive oxygen species and nitric oxide. FEBS Lett 489:237–242
McAinsh MR, Clayton H, Mansfield TA, Hetherington AM (1996) Changes in stomatal behavior and guard cell cytosolic free calcium in response to oxidative stress. Plant Physiol 111:1031–1042
Mori IC, Schroeder JI (2004) Reactive oxygen species activation of plant Ca2+ channels. A signaling mechanism in polar growth, hormone transduction, stress signaling, and hypothetically mechanotransduction. Plant Physiol 135:702–708
Neill SJ, Desikan R, Hancock JT (2002) Hydrogen peroxide signalling. Curr Opin Plant Biol 5:388–395
Neves C, Sá MC, Amâncio S (1998) Histochemical detection of H2O2 by tissue printing as a precocious marker of rhizogenesis in grapevine. Plant Physiol Biochem 36:817–824
Noctor G, Foyer CH (1998) Ascorbate and glutathione: keeping active oxygen under control. Ann Rev Plant Physiol Plant Mol Biol 49:249–279
Orozco-Cardenas M, Ryan CA (1999) Hydrogen peroxide is generated systemically in plant leaves by wounding and systemin via the octadecanoid pathway. Proc Natl Acad Sci USA 96:6553–6557
Pagnussat GC, Simontacchi M, Puntarulo S, Lamattina L (2002) Nitric oxide is required for root organogenesis. Plant Physiol 129:954–956
Pagnussat GC, Lanteri ML, Lorenzo Lamattina L (2003) Nitric oxide and cyclic GMP are messengers in the indole acetic acid-induced adventitious rooting process. Plant Physiol 132:1241–1248
Pagnussat GC, Lanteri ML, Lombardo MC, Lamattina L (2004) Nitric oxide mediates the indole acetic acid induction activation of a mitogen-activated protein kinase cascade involved in adventitious root development. Plant Physiol 135:279–286
Pei Z-M, Murata Y, Benning G, Thomine S, Klüsener B, Allen GJ, Grill E, Schroeder JI (2000) Calcium channels activated by hydrogen peroxide mediate abscisic acid signaling in guard cells. Nature 406:731–734
Price AH, Taylor A, Ripley SJ, Griffiths A, Trewavas AJ, Knight MR (1994) Oxidative signals in tobacco increase cytosolic calcium. Plant Cell 6:1301–1310
Puntarulo S, Sánchez RA, Boveris A (1988) Hydrogen peroxide metabolism in soybean axes at the onset of germination. Plant Physiol 86:626–630
Rentel MC, Lecourieux D, Ouaked F, Usher SL, Petersen L, Okamoto H, Knight H, Peck SC, Grierson CS, Hirt H, Knight MR (2004) OXI1 kinase is necessary for oxidative burst-mediated signalling in Arabidopsis. Nature 427:858–861
Rodriguez AA, Grunberg KA, Taleisnik EL (2002) Reactive oxygen species in the elongation zone of maize leaves are necessary for leaf extension. Plant Physiol 129:1627–1632
Rudd JJ, Franklin-Tong VE (2001) Unravelling response-specificity in Ca2+ signalling pathways in plant cells. New Phytol 151:7–33
Sanders D, Brownlee C, Harper JF (1999) Communicating with calcium. Plant Cell 11:691–706
Schopfer P, Plachy C, Frahry G (2001) Release of reactive oxygen intermediates (superoxide radicals, hydrogen peroxide, and hydroxyl radicals) and peroxidase in germinating radish seeds controlled by light, gibberellin, and abscisic acid. Plant Physiol 125:1591–1602
Schopfer P, Liszkay A, Bechtold M, Frahry G, Wagner A (2002) Evidence that hydroxyl radicals mediate auxin-induced extension growth. Planta 214:821–828
Su GX, Zhang WH, Liu YL (2006) Involvement of hydrogen peroxide generated by polyamine oxidative degradation in the development of lateral roots in soybean. J Integra Plant Biol 48:426–432
Suzuki Y, Yanagisawa H (1980) Purification and properties of maize polyamine oxidase: a flavoprotein. Plant Cell Physiol 21:1085–1094
Thordal-Christensen H, Zhang Z, Wei Y, Collinge DB (1997) Subcellular localization of H2O2 in plants: H2O2 accumulation in papillae and hypersensitive response during the barley-powdery mildew interaction. Plant J 11:1187–1194
Van Gestelen P, Asard H, Caubergs RJ (1997) Solubilization and separation of a plant plasma membrane NADPH-O2 synthase from other NAD(P)H oxidoreductases. Plant Physiol 115:543–550
Xing T, Higgins VJ, Blumwald E (1997) Race-specific elicitors of Cladosporium fulvum promote translocation of cytosolic components of NADPH oxidase to the plasma membrane of tomato cells. Plant Cell 9:249–259
Yoda H, Yamaguchi Y, Sano H (2003) Induction of hypersensitive cell death by hydrogen peroxide produced through polyamine degradation in tobacco plants. Plant Physiol 132:1973–1981
Yoshioka H, Numata N, Nakajima K, Katou S, Kawakita K, Rowland O, Jones JDG, Doke N (2003) Nicotiana benthamiana gp91phox homologs NbrbohA and NbrbohB participate in H2O2 accumulation and resistance to Phytophthora infestans. Plant Cell 15:706–718
Acknowledgments
This work was financially supported by National Natural Science Foundation of China (31000130), Shaanxi Provincial Natural Science Foundation for the Youth of China (2009JM3001) and the Fundamental Research Funds for the Central Universities (GK200902037).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Huang, AX., She, XP., Cao, BH. et al. Distribution of hydrogen peroxide during adventitious roots initiation and development in mung bean hypocotyls cuttings. Plant Growth Regul 64, 109–118 (2011). https://doi.org/10.1007/s10725-010-9545-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10725-010-9545-y