The experiment was carried out in the laboratory of tissue culture of Plant Production Techniques Department, at Musaib Technical College to study the effect of different concentrations of putrescine and the type of light on the content of the Gardenia from some effective medical compounds. The experiment included two factor the first consisted of two sources of light, namely light (fluorescent), LED light (18 red: 2 blue) and the second factor, the addition of Putrescine with three concentrations (0.5, 1 and 1.5) mg.l-1. Some effective medical substances (Coumaric, Ferulic, Caffeorylquanic, Sinapic acid and Tannic) acids was measured in Callus Of Gardenia using high-performance liquid chromatography (HPLC).The results were analyzed using the statistical program Genstat and the mean was measured according to the least significant difference of LSD at the probability level of 5%. The results showed superior treatment of LED lighting in the concentration of all measured active compounds, The highest concentration of compounds (Ferulic, Caffeorylquanic and Sinapic) acids was given with a concentration of 0.5 mg.l-1. The highest concentration of compounds (Coumaric and Tannic) acids. The interaction (LED + Putrescine at 0.5) gave the highest concentration of compounds (Ferulic, Caffeorylquanic and Sinapic) in Callus of Gardenia.