NADPH oxidase AtrbohD an d AtrbohF negatively modulate lateral root development by changing the peroxidase activity and increasing the local generation of superoxide in primary roots of Arabidopsis in an auxin-independent manner.
NADPH oxidase subunits AtrbohD and AtrbohF play pivotal roles in regulating growth, development and stress responses in Arabidopsis. However, whether they modulate lateral root (LR) formation has not yet been addressed, and the detailed mechanisms underlying the process remain unanswered. Here, we show that two null double mutants atrbohD1/F1 and atrbohD2/F2, in which both AtrbohD and AtrbohF genes are disrupted, had remarkably higher LR density than wild-type (WT), or the single mutant atrbohD1 and atrbohF1. Compared to WT, the double mutants exhibited early emerged LRs and enhanced density of lateral root primordia (LRP). Unexpectedly, the production of superoxide (O2 −), but not hydrogen peroxide, in the mature area of the primary root containing LRs significantly increased in the double mutants relative to that in WT. Further experiments revealed that the local accumulation of O2 − led to the enhancement of LR density in the double mutants. Moreover, the deficiency of AtrbohD and AtrbohF caused a marked increase in peroxidase activity in the mature root zone, which contributed to the localized accumulation of O2 − and the elevated LR density in the double mutants. Furthermore, the double mutants were not sensitive to exogenous auxin naphthalene acetic acid or auxin transport inhibitor 1-N-naphthylphthalamic acid in terms of LR formation. The auxin response of LRP in vivo in atrbohD1/F1 was also similar to that in WT. Taken together, these results suggest that AtrbohD and AtrbohF negatively modulate LR development by controlling the local generation of superoxide in an auxin-independent manner. These findings provide new insights into the mechanisms of NADPH oxidase-mediated regulation of LR branching in Arabidopsis.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Lateral root primordia
Reactive oxygen species
- H2DCFDA 2:
Naphthalene acetic acid
Bashandy T, Guilleminot J, Vernoux T, Caparros-Ruiz D, Ljung K, Meyer Y, Reichheld JP (2010) Interplay between the NADP-linked thioredoxin and glutathione systems in Arabidopsis auxin signaling. Plant Cell 22:376–391
Baxter A, Mittler R, Suzuki N (2014) ROS as key players in plant stress signalling. J Exp Bot 65:1229–1240
Casimiro I, Beeckman T, Graham N, Bhalerao R, Zhang H, Casero P, Sandberg G, Bennett MJ (2003) Dissecting Arabidopsis lateral root development. Trends Plant Sci 8:165–171
Chen YH, Chao YY, Hsu YY, Kao CH (2013) Heme oxygenase is involved in H2O2-induced lateral root formation in apocynin-treated rice. Plant Cell Rep 32:219–226
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
Foreman J, Demidchik V, Bothwell JHF, Mylona P, Miedema H, Torresk MA, Linstead P, Costa S, Brownlee C, Jonesk JDG, Davies JM, Dolan L (2003) Reactive oxygen species produced by NADPH oxidase regulate plant cell growth. Nature 422:442–446
Fukaki H, Tasaka M (2009) Hormone interactions during lateral root formation. Plant Mol Biol 69:437–449
Jiao YH, Sun LR, Song YL, Wang LM, Liu LP, Zhang LY, Liu B, Li N, Miao C, Hao FS (2013) AtrbohD and AtrbohF positively regulate abscisic acid inhibited primary root growth by affecting Ca2+ signaling and auxin response of roots in Arabidopsis. J Exp Bot 64:4183–4192
Kaur G, Sharma A, Guruprasad K, Pati PK (2014) Versatile roles of plant NADPH oxidases and emerging concepts. Biotechnol Adv. doi:10.1016/j.biotechadv.2014.02.002
Kim MJ, Ciani S, Schachtman DP (2010) A peroxidase contributes to ROS production during Arabidopsis root response to potassium deficiency. Mol Plant 3:420–427
Kwak JM, Mori IC, Pei ZM, Leonhardt N, Torres MA, Dangl JL, Bloom RE, Bodde S, Jones JD, Schroeder JI (2003) NADPH oxidase AtrbohD and AtrbohF genes function in ROS-dependent ABA signaling in Arabidopsis. EMBO J 22:2623–2633
Kwak JM, Ngyuen V, Schroeder JI (2006) The role of reactive oxygen species in hormonal responses. Plant Physiol 141:323–329
Lavenus J, Goh T, Roberts I, Guyomarc’h S, Lucas M, De Smet I, Fukaki H, Beeckman T, Bennett M, Laplaze L (2013) Lateral root development in Arabidopsis: fifty shades of auxin. Trends Plant Sci 18:450–458
Ma LY, Zhang H, Sun LR, Jiao YH, Zhang GZ, Miao C, Hao FS (2012) NADPH oxidase AtrbohD and AtrbohF function in ROS-dependent regulation of Na+/K+ homeostasis in Arabidopsis under salt stress. J Exp Bot 63:305–317
Malamy JE, Benfey PN (1997) Organization and cell differentiation in lateral roots of Arabidopsis thaliana. Development 124:33–44
Manzano C, Pallero-Baena M, Casimiro I, Rybel BD, Orman-Ligeza B, Isterdael GV, Beeckman T, Draye X, Casero P, Pozo JCD (2014) The emerging role of reactive oxygen species signaling during lateral root development. Plant Physiol 165:1105–1119
Marino D, Dunand C, Puppo A, Pauly N (2012) A burst of plant NADPH oxidases. Trends Plant Sci 17:9–15
Montiel J, Nava N, Cárdenas L, Sánchez-López R, Manoj-Kumar A, Santana O, Sánchez F, Quinto C (2012) A Phaseolus vulgaris NADPH oxidase gene is required for root infection by Rhizobia. Plant Cell Physiol 53:1751–1767
Müller K, Linkies A, Leubner-Metzger G, Kermode AR (2012) Role of a respiratory burst oxidase of Lepidium sativum (cress) seedlings in root development and auxin signaling. J Exp Bot 63:6325–6334
Nibau C, Gibbs DJ, Coates JC (2008) Branching out in new directions: the control of root architecture by lateral root formation. New Phytol 179:595–614
Olmos E, Kiddle G, Pellny TK, Kumar S, Foyer CH (2006) Modulation of plant morphology, root architecture, and cell structure by low vitamin C in Arabidopsis thaliana. J Exp Bot 57:1645–1655
Overvoorde P, Fukaki H, Beeckman T (2010) Auxin control of root development. Cold Spring Harb Perspect Biol 2:a001537
Owusu-Ansah E, Yavari A, Banerjee U (2008) A protocol for in vivo detection of reactive oxygen species. Nat Protoc http://www.natureprotocols.com/2008/02/27/a_protocol_for_in_vivo_detecti.php.10.1038/nprot.2008.23
Passaia G, Queval G, Bai J, Margis-Pinheiro M, Foyer CH (2014) The effects of redox controls mediated by glutathione peroxidases on root architecture in Arabidopsis thaliana. J Exp Bot 65:1403–1413
Passardi F, Tognolli M, De Meyer M, Penel C, Dunand C (2006) Two cell wall associated peroxidases from Arabidopsis influence root elongation. Planta 223:965–974
Pasternak T, Potters G, Caubergs R, Jansen MAK (2005) Complementary interactions between oxidative stress and auxins control plant growth responses at plant, organ, and cellular level. J Exp Bot 56:1991–2001
Péret B, De Rybel B, Casimiro I, Benková E, Swarup R, Laplaze L, Beeckman T, Bennett MJ (2009) Arabidopsis lateral root development: an emerging story. Trends Plant Sci 14:399–408
Potikha TS, Collins CC, Johnson DI, Delmer DP, Levine A (1999) The involvement of hydrogen peroxide in the differentiation of secondary walls in cotton fibers. Plant Physiol 119:849–858
Shin R, Berg RH, Schachtman DP (2005) Reactive oxygen species and root hairs in Arabidopsis root response to nitrogen, phosphorus and potassium deficiency. Plant Cell Physiol 46:1350–1357
Song CP, Agarwal M, Ohta M, Guo Y, Halfter U, Wang P, Zhu JK (2005) Role of an Arabidopsis AP2/EREBP-type transcriptional repressor in abscisic acid and drought stress responses. Plant Cell 17:2384–2396
Srinivasan VS, Podolski D, Westrick NJ, Neckers DC (1978) Photochemical generation of superoxide (O2 −) by Rose Bengal and Ru(Bpy)32+. J Am Chem Soc 100:6513–6515
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 Integr Plant Biol 48:426–432
Thordal-Christensen H, Zhang ZG, Wei YD, 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
Tognetti VB, Mühlenbock P, Breusegem FV (2012) Stress homeostasis–the redox and auxin perspective. Plant, Cell Environ 35:321–333
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, Dunajska-Ordak K, Skorupa M, Tretyn A (2012) Role of ascorbate in the regulation of the Arabidopsis thaliana root growth by phosphate availability. J Bot. doi:10.1155/2012/580342
Zhang H, Jennings A, Barlow PW, Forde GB (1999) Dual pathways for regulation of root branching by nitrate. Proc Natl Acad Sci USA 96:6529–6534
This work was supported by the National Natural Science Foundation of China [31070239 and 30970235], and by the Support Plan for Talents in Innovation of Science and Technology in Henan Province [2011HASTIT007]. We thank Dr. Jianwei Pan (Zhejiang Normal University in China) for kindly providing the seeds of the ProDR5:GFP transgenic Arabidopsis line.
N. Li and L. Sun contributed equally to this work.
Electronic supplementary material
Below is the link to the electronic supplementary material.
About this article
Cite this article
Li, N., Sun, L., Zhang, L. et al. AtrbohD and AtrbohF negatively regulate lateral root development by changing the localized accumulation of superoxide in primary roots of Arabidopsis . Planta 241, 591–602 (2015). https://doi.org/10.1007/s00425-014-2204-1
- Lateral root formation
- NADPH oxidase