Gravistimulation induces the rapid accumulation of NO required for gravitropism in soybean roots. a, Kinetics of NO accumulation in Zone 1 (•) or Zone 2 (○) from horizontal roots. ▾, Vertical roots. b, Correlation between NO accumulation in Zone 1 (black bars) and gravitropic curvature (white bars). Roots were pretreated as indicated for 12 h and then placed horizontally for 12 h (50 μm cPTIO; 20 μm l-NAA and NaN3; 30 μm PBITU and NPA; 100 μm ODQ and LY83583). On x axis: C, Control; G, gravistimulated. Values are the mean ± se for five independent experiments. Data analyzed by one-way ANOVA followed by Tukey's test. Different symbols indicate significant differences between treatments (P < 0.05). c, Gravistimulation induces asymmetric NO accumulation. Soybean roots were loaded with DAF-2DA and gravistimulated by orientating horizontally. Fluorescence intensity of dissected root tips was observed at the indicated times by confocal fluorescence microscopy. Effects of pretreatment with 20 μm NaN3, 20 μm l-NNA, 50 μm cPTIO, and 10 μm NPA are shown. The pictures were taken after 6 h treatment. Scale bar = 1 mm. Experiments were repeated at least five times with similar results. d, Asymmetric accumulation of NO in root Zone 1. Soybean roots were gravistimulated for different times, bisected horizontally with a razor blade, and NO content assayed. ▾, Vertical roots; ○, lower half of horizontal root; •, upper half of horizontal root.