Optimization of Direct Shoot Regeneration Using Leaf Explants From Cichorium Intybus By Response Surface Methodology

Medicinal plants play an important role in treatments of ailments with fewer side effects. Chicory (Cichorium intybus), a typical coffee substitute, is also a medicinally important plant used to treat ailments ranging from jaundice to tumors and cancer. The levels of growth factors and conditions (number of days, sucrose concentration, BAP concentration and kinetin concentration) which affect the shoot generation from the leaf explants of C. intybus were obtained by one factor at a time which was further optimized by a central composite design using response surface methodology. A second order quadratic model was generated, which was validated and was found to be of good fit, with an R2 value of 0.8928. At the optimized conditions of 20 g/L sucrose, 0.75 mg/L BAP and 0.3 mg/L kinetin at 30 days of growth, largest number of 20 shoots were produced, which gave a 1.38 fold increase in comparison to the un-optimized conditions. These results indicate that response surface methodology offers an efficient methodology to identify and optimize the significant factors with a minimum number of experiments for plant regeneration of C. intybus and related species in vitro.