Secondary metabolites production from liquid tissue culture ofCitrus aurantium

This work focused on selection of theCitrus aurantiumexplant and hormonal combination that gives cells with high production level of secondary metabolites in liquid medium.The highest percentage of callus formation (100%) was observed for cotyledonary with 3mg / l 2,4-D and 2 mg / l kinetin and with ovules with 0.2 mg/l 2,4-D and 2 mg/l kinetin.Analyses of essential oil composition by gas chromatography revealed that neither callus from cotyledonary nor from ovules explants product volatiles components.Analysis of callus fatty acid revealed that those produced by calluses issued from ovules cultures are those that are closest to those formed in the bitter orange fruit.Transfer of calluses derived from ovules and cotyledonaries explants gives two types of suspension depending on the hormonal combination. Calluses transferred on Liquid MS medium supplemented with 0.2 mg/l 2,4-D with 2 mg/l kinetin gives a dispersed suspension. Howether, calluses transferred onto MS liquid medium supplemented 4 mg/l BA and 0.2 mg/ NAA gives suspension with compact cluster callus.The growth rate of callus aggregate was lower than the dispersed cell cultures and their biomass were higher than the dispersed cell cultures.Transfer of calluses in liquid medium show that cytokinins at high concentrations are essential for initiation and maintenance of callus aggregate.