Nitric oxide accumulation and actin distribution during auxin-induced adventitious root development in sunflower

Auxin-mediated adventitious root (AR) formation from the basal ends of hypocotyl segments in sunflower (Helianthus annuus L, cv. Morden) is associated with gradual subcellular and anatomical changes. Pre-mitotic events (induction phase) are evident within 1 d (24 h) in the interfascicular parenchyma. Evidence for nitric oxide (NO) accumulation in the interfascicular cells 2d after indole-3-acetic acid (IAA) treatment, highlights the involvement of NO specifically during root initiation, following AR induction phase. Thus, adventitious root induction and initiation can be distinguished as NO-independent and NO-dependent phases. Treatment of hypocotyl explants with 10 mu M 1-napthylphthlamic acid (NPA), an inhibitor of polar auxin transport in plant cells, leads to complete abolition of NO-associated fluorescence, detected upon incubation of hypocotyl sections with 4,5-diaminofluorescein diacetate (DAF-2DA). This indicates a signaling link between auxin transport and NO accumulation in the target cells. Latrunculin B (Lat B), an actin depolymerizing agent, leads to substantial inhibition of IAA-induced AR response and NO accumulation in the responding cells. Aminoguanidine [an inhibitor of inducible form of nitric oxide synthase (iNOS)] brings about a reduction in the tissue NO content, indicating a significant activity of putative NOS leading to endogenous NO accumulation. Confocal laser scanning microscope (CLSM) imaging has revealed bundling of actin with NPA and Lat B treatments. A probable interaction is thus evident between actin and NO during auxin-mediated AR formation in the initiation and extension phase. Present work, thus, provides novel observations on an interaction among endogenous NO. IAA and actin at specific stages of AR formation. (C) 2011 Elsevier B.V. All rights reserved.