L-Arginine optimization medium for hairy root induction of madder (Rubia tinctorum L.) based on one factor model of response surface methodology.

Medium optimization for hairy root cultures producing secondary metabolites was studied through statistical experimental design. In the following, one factor model of response surface methodology (RSM) was employed to formulate the L-Arginine amino acid levels alongside three categorical factors including bacterial strains (ATCC 15834, C58C1 and R1000), type of explant (Leaf and Stem) in co-cultivation media (B5 and MS) for hairy root induction of Madder ( Rubia tinctorum L.). Design of experiment and data analysis was carried out by using Expert-Design ® 7.1 software. According to the results, modeling and optimization conditions, including L-Arginine concentration 1.00 mM; bacterial strain; C58C1, leaf explant and B5 medium for HR induction frequency equal to 58% was evaluated (Desirability point=0.986). These optimal conditions predicted by RSM were confirmed to enhance hairy root induction as an application potential for biotechnological implementation to produce the anticipated compounds.