NO has a role in shoot regeneration. (A) NO staining at the border of leaf segments placed on regeneration medium in the presence of NO inhibitor (Reg+I), NO donor (Reg+D) and control (Reg). (B) Stain intensity per cell at the borders of leaf segments during the induction period of shoot induction. NO inhibitor (Regd), NO donor (Regm), and control (Reg). (C) Effect of NO donors (S-Nitroso-N-acetyl-DL-penicillamine; Molsidomine), NO scavenger (PTIO), and NO synthesis inhibitor (Diphenyleneiodonium) on shoot regeneration. Different letters (A, B, C) define statistically different results, p(f) was 7.65 × 10−35 using Tukey test. (D) Transcript level of Nitric Oxide Synthase1 increases during shoot induction while transcript level of Nitric Oxide Synthase2 is unchanged. The data presented are from the Differential Gene Expression experiment. (E) Transcript level of all Nitrate reductase genes increase during incubation on Reg medium. The data presented are from the Differential Gene Expression experiment. Reg = regeneration medium; regm = regeneration medium with added Molsidomine; regd = regeneration medium with added Diphenyleneiodonium. NR = nitrate reductase; NOS = nitric oxide synthase. Error bars indicated standard error of 50 cells in (B); 20 segment in 3 replicated in (C); and 3 biological replicates in (D,E).