, Volume 231, Issue 5, pp 1025–1036 | Cite as

Nitric oxide mediates humic acids-induced root development and plasma membrane H+-ATPase activation

  • Daniel B. Zandonadi
  • Mirella P. Santos
  • Leonardo B. Dobbss
  • Fábio L. Olivares
  • Luciano P. Canellas
  • Marla L. Binzel
  • Anna L. Okorokova-Façanha
  • Arnoldo R. Façanha
Original Article


It is widely reported that some humic substances behave as exogenous auxins influencing root growth by mechanisms that are not yet completely understood. This study explores the hypothesis that the humic acids’ effects on root development involve a nitric oxide signaling. Maize seedlings were treated with HA 20 mg C L−1, IAA 0.1 nM, and NO donors (SNP or GSNO), in combination with either the auxin-signaling inhibitor PCIB, the auxin efflux inhibitor TIBA, or the NO scavenger PTIO. H+-transport-competent plasma membrane vesicles were isolated from roots to investigate a possible link between NO-induced H+-pump and HA bioactivity. Plants treated with either HA or SNP stimulated similarly the lateral roots emergence even in the presence of the auxin inhibitors, whereas NO scavenger diminished this effect. These treatments induced H+-ATPase stimulation by threefold, which was abolished by PTIO and decreased by auxin inhibitors. HA-induced NO synthesis was also detected in the sites of lateral roots emergence. These data depict a new scenario where the root development stimulation and the H+-ATPase activation elicited by either HA or exogenous IAA depend essentially on mechanisms that use NO as a messenger induced site-specifically in the early stages of lateral root development.


Auxin Humic substances Lateral roots Nitric oxide pH Proton pump 



Indole-3-acetic acid


Humic acids


Nitric oxide


Lateral root


Sodium nitroprusside


4,5-Diaminofluorescein diacetate


Humic substances


p-Chlorophenoxyisobutyric acid


2,3,5-Triiodobenzoic acid




Plasma membrane


Sodium orthovanadate




Primary root


Initial velocity


Fluorescence quenching




2-(N-Morpholino)ethanesulfonic acid




4-Morpholinepropanesulfonic acid



This work was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) and International Foundation for Science (IFS).


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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Daniel B. Zandonadi
    • 1
  • Mirella P. Santos
    • 2
  • Leonardo B. Dobbss
    • 2
  • Fábio L. Olivares
    • 1
  • Luciano P. Canellas
    • 2
  • Marla L. Binzel
    • 3
  • Anna L. Okorokova-Façanha
    • 1
  • Arnoldo R. Façanha
    • 1
  1. 1.Centro de Biociências e Biotecnologia (CBB)Universidade Estadual do Norte Fluminense Darcy Ribeiro (UENF)Rio de JaneiroBrazil
  2. 2.Centro de Ciências e Tecnologias Agropecuárias (CCTA)UENFRio de JaneiroBrazil
  3. 3.Texas AgriLife ResearchDallasUSA

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