Nitric oxide causes root apical meristem defects and growth inhibition while reducing PIN-FORMED 1 (PIN1)-dependent acropetal auxin transport.
Nitric oxide (NO) is considered a key regulator of plant developmental processes and defense, although the mechanism and direct targets of NO action remain largely unknown. We used phenotypic, cellular, and genetic analyses in Arabidopsis thaliana to explore the role of NO in regulating primary root growth and auxin transport. Treatment with the NO donors S-nitroso-N-acetylpenicillamine, sodium nitroprusside, and S-nitrosoglutathione reduces cell division, affecting the distribution of mitotic cells and meristem size by reducing cell size and number compared with NO depletion by 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO). Interestingly, genetic backgrounds in which the endogenous NO levels are enhanced [chlorophyll a/b binding protein underexpressed 1/NO overproducer 1 (cue1/nox1) mirror this response, together with an increased cell differentiation phenotype. Because of the importance of auxin distribution in regulating primary root growth, we analyzed auxin-dependent response after altering NO levels. Both elevated NO supply and the NO-overproducing Arabidopsis mutant cue1/nox1 exhibit reduced expression of the auxin reporter markers DR5pro:GUS/GFP. These effects were accompanied by a reduction in auxin transport in primary roots. NO application and the cue1/nox1 mutation caused decreased PIN-FORMED 1 (PIN1)-GFP fluorescence in a proteasome-independent manner. Remarkably, the cue1/nox1-mutant root phenotypes resemble those of pin1 mutants. The use of both chemical treatments and mutants with altered NO levels demonstrates that high levels of NO reduce auxin transport and response by a PIN1-dependent mechanism, and root meristem activity is reduced concomitantly.
Department of Fisiología Vegetal, Centro Hispano-Luso de Investigaciones Agrarias, Facultad de Biología,  Universidad de Salamanca, 37185 Salamanca,  Spain.
María
M
Fernández-Marcos
Luis
L
Sanz
Daniel R
DR
Lewis
Gloria K
GK
Muday
Oscar
O
Lorenzo
D017360:Arabidopsis / Q000254:growth & development; D029681:Arabidopsis Proteins / Q000502:physiology*; D001692:Biological Transport; D007210:Indoleacetic Acids / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000502:physiology*; D018519:Meristem / Q000254:growth & development*; D009569:Nitric Oxide / Q000502:physiology*; D020030:Nitric Oxide Donors / Q000008:administration & dosage; D009599:Nitroprusside / Q000008:administration & dosage; D018517:Plant Roots / Q000254:growth & development*; D060888:Real-Time Polymerase Chain Reaction
D029681:Arabidopsis Proteins; D007210:Indoleacetic Acids; D026901:Membrane Transport Proteins; D020030:Nitric Oxide Donors; C116363:PIN1 protein, Arabidopsis; D009599:Nitroprusside; D009569:Nitric Oxide