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      RbohA coordinates lateral root emergence in common bean

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          ABSTRACT

          Respiratory burst oxidase homologs (RBOHs) constitute a multigene family in plants. These reactive oxygen species (ROS)-generating enzymes participate in diverse biological processes. We previously demonstrated that PvRbohB plays an important role in lateral root (LR) development in Phaseolus vulgaris. However, little is known about the roles of other Rboh members in root development. Here, we report that PvRbohA is involved in LR emergence in P. vulgaris. PvRbohA was expressed in wild-type P. vulgaris root tissues, e.g., the radicle, inter LR zone, and LR zone, and its expression increased during LR formation. Analysis of the spatio-temporal expression patterns of a reporter construct under the control of the PvRbohA promoter ( PvRbohA::GUS-GFP) in transgenic roots showed that PvRbohA was active at all three stages of LR development, but its spatial expression pattern varied at each stage. The relative expression levels of PvRbohA during LR formation correlated with the activity of PvRbohA promoter. Furthermore, upon PvRbohA transcript silencing, LR growth was significantly altered in transgenic hairy roots. These findings suggest that RbohA participates in LR initiation, emergence, and development in the legume P. vulgaris by delimiting the region for LR emergence.

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          Assaying chimeric genes in plants: The GUS gene fusion system

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            Reactive oxygen species produced by NADPH oxidase regulate plant cell growth.

            Cell expansion is a central process in plant morphogenesis, and the elongation of roots and root hairs is essential for uptake of minerals and water from the soil. Ca2+ influx from the extracellular store is required for (and sets the rates of) cell elongation in roots. Arabidopsis thaliana rhd2 mutants are defective in Ca2+ uptake and consequently cell expansion is compromised--rhd2 mutants have short root hairs and stunted roots. To determine the regulation of Ca2+ acquisition in growing root cells we show here that RHD2 is an NADPH oxidase, a protein that transfers electrons from NADPH to an electron acceptor leading to the formation of reactive oxygen species (ROS). We show that ROS accumulate in growing wild-type (WT) root hairs but their levels are markedly decreased in rhd2 mutants. Blocking the activity of the NADPH oxidase with diphenylene iodonium (DPI) inhibits ROS formation and phenocopies Rhd2-. Treatment of rhd2 roots with ROS partly suppresses the mutant phenotype and stimulates the activity of plasma membrane hyperpolarization-activated Ca2+ channels, the predominant root Ca2+ acquisition system. This indicates that NADPH oxidases control development by making ROS that regulate plant cell expansion through the activation of Ca2+ channels.
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              NADPH oxidase AtrbohD and AtrbohF genes function in ROS-dependent ABA signaling in Arabidopsis.

              Reactive oxygen species (ROS) have been proposed to function as second messengers in abscisic acid (ABA) signaling in guard cells. However, the question whether ROS production is indeed required for ABA signal transduction in vivo has not yet been addressed, and the molecular mechanisms mediating ROS production during ABA signaling remain unknown. Here, we report identification of two partially redundant Arabidopsis guard cell-expressed NADPH oxidase catalytic subunit genes, AtrbohD and AtrbohF, in which gene disruption impairs ABA signaling. atrbohD/F double mutations impair ABA-induced stomatal closing, ABA promotion of ROS production, ABA-induced cytosolic Ca(2+) increases and ABA- activation of plasma membrane Ca(2+)-permeable channels in guard cells. Exogenous H(2)O(2) rescues both Ca(2+) channel activation and stomatal closing in atrbohD/F. ABA inhibition of seed germination and root elongation are impaired in atrbohD/F, suggesting more general roles for ROS and NADPH oxidases in ABA signaling. These data provide direct molecular genetic and cell biological evidence that ROS are rate-limiting second messengers in ABA signaling, and that the AtrbohD and AtrbohF NADPH oxidases function in guard cell ABA signal transduction.
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                Author and article information

                Journal
                Commun Integr Biol
                Commun Integr Biol
                KCIB
                kcib20
                Communicative & Integrative Biology
                Taylor & Francis
                1942-0889
                2018
                11 May 2018
                11 May 2018
                : 11
                : 2
                : 1-5
                Affiliations
                [a ]Ciencias Agrogenómicas, Escuela Nacional de Estudios Superiores Unidad León, Universidad Nacional Autónoma de México (UNAM), León , Guanajuato, México
                [b ]Departamento de Biología Molecular de Plantas, Instituto de Biotecnología, Universidad Nacional Autónoma de México (UNAM) , Cuernavaca, Morelos, México
                Author notes
                CONTACT Dr. Carmen Quinto quinto@ 123456ibt.unam.mx Departamento de Biología Molecular de Plantas, Instituto de Biotecnología, Universidad Nacional Autónoma de México, UNAM , Apartado Postal 510-3, Cuernavaca 62271, Morelos, Mexico

                Addendum for: Arthikala, M.-K., Montiel, J., Sánchez-López, R., Nava, N., Cárdenas, L., & Quinto, C. (2017). Respiratory Burst Oxidase Homolog Gene A Is Crucial for Rhizobium Infection and Nodule Maturation and Function in Common Bean. Frontiers in Plant Science, 8, 2003. http://doi.org/10.3389/fpls.2017.02003

                Article
                1467188
                10.1080/19420889.2018.1467188
                6067902
                4adcac13-e574-4ab3-b162-4ab4a98dd84b
                © 2018 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 24 February 2018
                : 10 April 2018
                : 12 April 2018
                Page count
                Figures: 3, Tables: 0, Equations: 0, References: 20, Pages: 5
                Funding
                This work was partially supported by Consejo Nacional de Ciencia y Tecnología (CB-2010-153718 to C.Q) with a postdoctoral fellowship (17656) to M-K.A., and by DGAPA IN224115 to C.Q. DGAPA/PAPIIT-UNAM grant no. IA205117 to M.-K.A.
                Categories
                Article Addendum

                Molecular biology
                common bean,lateral root emergence,nadph oxidase,promoter rboha,rboha-rnai
                Molecular biology
                common bean, lateral root emergence, nadph oxidase, promoter rboha, rboha-rnai

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