Abstract
Background and aims
The aim of this study was to test the effect of Azospirillum brasilense on the superoxide anion production (O2 •−) and enzymes related with redox metabolism in roots of wheat (Triticum aestivum).
Methods
A. brasilense Sp245 and T. aestivum seeds cv Nana F2007 were used in this study. Wheat roots were stained with nitro blue tetrazolium (NBT) to visualize and localize O2 •− production. Superoxide dismutase (SOD) and peroxidase (POX) activities were assayed in native PAGE.
Results
We found that A. brasilense application resulted in a decrease in meristem length and cell size, and in a reduction in the O2 •− level in roots. The bacteria stimulated SOD and soluble POX isoenzymes, particularly in the zone of the root tip. Qualitative O2 •− production in roots treated with LaCl3, a Ca2+ channel blocker, in combination with A. brasilense was comparable to inoculated roots. Similar results were observed with the Ca2+ ionophore A23187.
Conclusions
Our results suggest that O2 •− metabolism is important during the interaction of wheat and A. brasilense, and that the antioxidative enzymes such as SOD and POX are involved in its regulation.
Similar content being viewed by others
References
Alquéres S, Meneses C, Rouws L, Rothballer M, Baldani I, Schmid M, Hartmann A (2013) The bacterial superoxide dismutase and glutathione reductase are crucial for endophytic colonization of rice roots by Gluconacetobacter diazotrophicus PAL5. Mol Plant-Microbe Interact 26:937–945
Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399
Arthikala M-K, Sanchez-Lopez R, Nava N, Santana O, Cardenas L, Quinto C (2014) RbohB, a Phaseolus vulgaris NADPH oxidase gene, enhances symbiosome number, bacteroid size, and nitrogen fixation in nodules and impairs mycorrhizal colonization. New Phytol 202:886–900
Baldani VLD, de B. Alvarez MA, Baldani JI, Dobereiner JD (1986) Establishment of inoculated Azospirillum spp in the rhizosphere and in roots of field grown wheat and sorghum. Plant Soil 90:35–46
Bashan Y, Holguin G, de-Bashan LE (2004) Azospirillum-plant relationships: physiological, molecular, agricultural, and environmental advances (1997– 2003). Can J Microbiol 50:521–577
Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44:276–287
Beyer WF, Fridovich I (1989) Characterization of a superoxide dismutase mimic prepared from desferrioxamine and MnO2. Arch Biochem Biophys 271:149–156
Bibikova TN, Zhigilei A, Gilroy S (1997) Root hair growth in Arabidopsis thaliana is directed by calcium and an endogenous polarity. Planta 203:495–505
Bottini R, Cassán F, Piccoli P (2004) Gibberellin production by bacteria and its involvement in plant growth promotion and yield increase. Appl Microbiol Biotechnol 65:497–503
Bradford MM (1976) A rapid and sensitive method for the quantization of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Bright J, Desikan R, Hancock JT, Weir IS, Neill SJ (2006) ABA-induced NO generation and stomatal closure in Arabidopsis are dependent on H2O2 synthesis. Plant J 45:113–122
Camilios-Neto D, Bonato P, Wassem R, Tadra-Sfeir MZ, Brusamarello-Santos LCC, Valdameri G, Donatti L, Faoro H, Weiss VA, Chubatsu LS, Pedrosa FO, Souza EM (2014) Dual RNA-seq transcriptional analysis of wheat roots colonized by Azospirillum brasilense reveals up-regulation of nutrient acquisition and cell cycle genes. BMC Genomics 15:378
Causin HF, Roqueiro G, Petrillo E, Láinez V, Pena LB, Marchetti CF, Gallego SM, Maldonado SI (2012) The control of root growth by reactive oxygen species in Salix nigra Marsh. Seedlings. Plant Sci 183:197–205
Corpas FJ, Barroso JB, del Río LA (2001) Peroxisomes as a source of reactive oxygen species and nitric oxide signal molecules in plant cells. Trends Plant Sci 6:145–150
Crozier A, Arruda P, Jasmim JM, Monteiro AM, Sandberg G (1988) Analysis of indole-3-acetic acid and related indoles in culture medium from Azospirillum lipoferum and Azospirillum brasilense. Appl Environ Microbiol 54:2833–2837
Dobbelaere S, Croonenborghs A, Thys A, Vande BA, Vanderleyden J (1999) Phytostimulatory effect of Azospirillum brasilense wild type and mutant strains altered in IAA production on wheat. Plant Soil 212:155–164
Dunand C, Crèvecoeur M, Penel C (2007) Distribution of superoxide and hydrogen peroxide in Arabidopsis root and their influence on root development: possible interaction with peroxidases. New Phytol 174:332–341
Ehrhardt DW, Wais R, Long SR (1996) Calcium spiking in plant root hairs responding to Rhizobium nodulation signals. Cell 85:673–681
Ellson CD, Gobert-Gosse S, Anderson KE, Davidson K, Erdjument-Bromage H, Tempst P, Thuring JW, Coope MA, Lim ZY, Holmes AB, Gaffney PR, Coadwell J, Chilvers ER, Hawkins PT, Stephens LR (2001) PtdIns(3)P regulates the neutrophil oxidase complex by binding to the PX domain of p40phox. Nat Cell Biol 3:679–682
Foreman J, Demidchik V, Bothwell JHF, Mylona P, Miedema H, Torres MA, Linstead P, Costa S, Brownlee C, Jones JDG, Davies JM, Dolan L (2003) Reactive oxygen species produced by NADPH oxidase regulate plant cell growth. Nature 422:442–446
Foyer CH, Noctor G (2009) Redox regulation in photosynthetic organisms: signaling, acclimation, and practical implications. Antioxid Red Signal 11:861–905
Geiger D, Maierhofer T, Al-Rasheid KA, Scherzer S, Mumm P, Liese A, Ache P, Wellmann C, Marten I, Grill E, Romeis T, Hedrich R (2011) Stomatal closure by fast abscisic acid signaling is mediated by the guard cell anion channel SLAH3 and the receptor RCAR1. Sci Signal 4:ra32
Ivanchenko MG, den Os D, Monhausen GB, Dubrovsky JG, Bednarova A, Krishnan N (2013) Auxin increases the hydrogen peroxide (H2O2) concentration in tomato (Solanum lycopersicum) root tips while inhibiting root growth. Ann Bot 112:1107–16
Jacoud C, Faure D, Wadoux P, Bally R (1998) Development of a strain-specific probe to follow inoculated Azospirillum lipoferum CRT1 under field conditions and enhancement of maize root development by inoculation. FEMS Microbiol Ecol 27:43–51
Joo JH, Yoo HJ, Hwang I, Lee JS, Nam KH, Bae YS (2005) Auxin-induced reactive oxygen species production requires the activation of phosphatidylinositol 3-kinase. FEBS Lett 579:1243–1248
Khalid A, Arshad M, Zahir ZA (2004) Screening plant growth-promoting rhizobacteria for improving growth and yield of wheat. J Appl Microbiol 96:473–480
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
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
Liszkay A, van der Zalm E, Schopfer P (2004) Production of reactive oxygen intermediates (O2 •-, H2O2, and •OH) by maize roots and their role in wall loosening and elongation growth. Plant Physiol 136:3114–3123
Lugtenberg B, Kamilova F (2009) Plant-growth-promoting rhizobacteria. Annu Rev Microbiol 63:541–556
Martinez-Morales LJ, Soto-Urzúc L, Baca BE, Sanchez-Ahécdo JA (2003) Indole-3-butyric acid (IBA) production in culture medium by wild strain Azospirillum brasilense. FEMS Microbiol Lett 228:167–173
Mittler R, Vanderauwera S, Gollery M, Van Breusegem F (2004) The reactive oxygen gene network in plants. Trends Plant Sci 9:490–498
Neill SJ, Desikan R, Clarke A, Hurst RD, Hancock JT (2002) Hydrogen peroxide and nitric oxide as signaling molecules in plants. J Exp Bot 53:1237–1247
Noctor G, Foyer CH (1998) Ascorbate and glutathione: keeping active oxygen under control. Annu Rev Plant Physiol Plant Mol Biol 49:249–279
Okon Y, Labandera-Gonzalez CA (1994) Agronomic applications of Azospirillum: an evaluation of 20 years worldwide field inoculation. Soil Biol Biochem 26:1591–1601
Peer WA, Cheng Y, Murphy AS (2013) Evidence of oxidative attenuation of auxin signalling. J Exp Bot 64:2629–2639
Perrig D, Boiero ML, Masciarelli OA, Penna C, Ruiz OA, Cassán FD, Luna MV (2007) Plant-growth-promoting compounds produced by two agronomically important strains of Azospirillum brasilense, and implications for inoculants formulation. Appl Microbiol Biotechnol 75:1143–1150
Potters G, Horemans N, Jansen MAK (2010) The cellular redox state in plant stress biology – a charging concept. Plant Physiol Biochem 48:292–300
Prigent-Combaret C, Blaha D, Pothier JF, Vial L, Poirier M-A, Wisniewski-Dyé F, Moënne-Loccoz Y (2008) Physical organization and phylogenetic analysis of acdR as leucine-responsive regulator of the 1-aminocyclopropane-1- carboxylate (ACC) deaminase gene acdS in phytobeneficial Azospirillum lipoferum 4B and other Proteobacteria. FEMS Microbiol Ecol 65:202–219
Raaijmakers JM, Paulitz TC, Steinberg C, Alabouvette C, Moenne-Loccoz Y (2009) The rhizosphere: a playground and battlefield for soil borne pathogens and beneficial microorganisms. Plant Soil 321:341–361
Renew S, Heyno E, Schopfer P, Liszkay A (2005) Sensitive detection and localization of hydroxyl radical production in cucumber roots and Arabidopsis seedlings by spin trapping electron paramagnetic resonance spectroscopy. Plant J 44:342–347
Rezzonico F, Binder C, Défago G, Moënne-Loccoz Y (2005) The type III secretion system of biocontrol Pseudomonas fluorescens KD targets the phytopathogenic chromista Pythium ultimum and promotes cucumber protection. Mol Plant-Microbe Interact 18:991–1001
Richardson AE, Barea JM, McNeill AM, Prigent-Combaret C (2009) Acquisition of phosphorus and nitrogen in the rhizosphere and plant growth promotion by microorganisms. Plant Soil 321:305–339
Spaepen S, Bossuyt S, Engelen K, Marchal K, Vanderleyden J (2014) Phenotypical and molecular responses of Arabidopsis thaliana roots as a result of inoculation with the auxin-producing bacterium Azospirillum brasilense. New Phytol 201:850–861
Steenhoudt O, Vanderleyden J (2000) Azospirillum, a free-living nitrogen-fixing bacterium closely associated with grasses: genetic, biochemical and ecological aspects. FEMS Microbiol Rev 24:487–506
Takeda S, Gapper C, Kaya H, Bell E, Kuchitsu K, Dolan L (2008) Local positive feedback regulation determines cell shape in root hair cells. Science 319:1241–1244
Tognetti VB, Per M, Frank VB (2012) Stress homeostasis – the redox and auxin perspective. Plant Cell Environ 35:321–333
Toyata M, Furuichi T, Tatsumi H, Sokabe M (2008) Critical consideration on the relationship between auxin transport and calcium transients in gravity perception of Arabidopsis seedlings. Plant Signal Behav 3:521–524
Tsavkelova EA, Klimova SY, Cherdyntseva TA, Netrusov AI (2006) Microbial producers of plant growth stimulators and their practical use: a review. Appl Biochem Microbiol 42:117–126
Tsavkelova EA, Cherdyntseva TA, Botina SG, Netrusov AI (2007) Bacteria associated with orchid roots and microbial production of auxin. Microbiol Res 162:69–76
Tsukagoshi H (2012) Defective root growth triggered by oxidative stress is controlled through the expression of cell cycle-related genes. Plant Sci 197:30–39
Tsukagoshi H, Busch W, Benfey PN (2010) Transcriptional regulation of ROS controls transition from proliferation to differentiation in the root. Cell 143:606–616
Tyburski J, Krzeminski L, Tretyn A (2009) Exogenous auxin affects ascorbato metabolism in roots of tomato seedlings. Plant Growth Regul 54:203–215
Van Breusegem F, Dat JF (2006) Reactive oxygen species in plant cell death. Plant Physiol 141:384–390
Vande Broek A, Michiels J, Van Gool A, Vanderleyden J (1993) Spatial-temporal colonization patterns of Azospirillum brasilense on the wheat root surface and expression of the bacterial nifH gene during association. Mol Plant-Microbe Interact 6:592–600
Vande Broek A, Dobbelaere S, Vanderleyden J, Van Dommeles A (2000) Azospirillum- plant root interactions: signaling and metabolic interactions. In: Triplett EW (ed) Prokariotic nitrogen fixation: a model system for the analysis of a biological process. Horizon Scientific Press, Wymondham, pp 761–777
Yang J, Kloepper JW, Ryu CM (2009) Rhizosphere bacteria help plants tolerate abiotic stress. Trends Plant Sci 14:1–4
Zamioudis C, Mastranesti P, Dhonukshe P, Blilou I, Pieterse CMJ (2013) Unraveling root developmental programs initiated by beneficial Pseudomonas spp. bacteria. Plant Physiol 162:304–318
Zelinová V, Halušková L, Mistrík I, Tamás L (2011) Abiotic stress–induced inhibition of root growth and ascorbic acid oxidase activity in barley root tip is associated with enhanced generation of hydrogen peroxide. Plant Soil 349:281–289
Zhao X, X-w Z, He H, Y-x W, Zhang X (2010) Mechanisms of extracellular NO and Ca2+ regulating the growth of wheat seedling roots. J Plant Biol 53:275–281
Acknowledgments
This study was supported by the Coordinación de la Investigación Científica, Universidad Michoacana de San Nicolás de Hidalgo, México.
Author information
Authors and Affiliations
Corresponding author
Additional information
Responsible Editor: Choong-Min Ryu.
Rights and permissions
About this article
Cite this article
Méndez-Gómez, M., Castro-Mercado, E., Alexandre, G. et al. Superoxide anion production in the interaction of wheat roots and rhizobacteria Azospirillum brasilense Sp245. Plant Soil 400, 55–65 (2016). https://doi.org/10.1007/s11104-015-2709-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11104-015-2709-9