Plant tissue or cell culture keeps a significant role in micro-propagation in the plant production industry. Combination of 6-Benzylaminopurine (BAP) and other plant growth regulators like 1-Naphthaleneacetic acid (NAA) or Indole-3-acetic acid (IAA) or indole-3-butyric acid (IBA) was used in the most of the research in tissue culture. The study was carried out to investigate the optimization of the concentration of IBA and BAP combination (0, 0.25, 0.50, 1.0, 1.50, 2.0, 2.5, 3.0 and 3.5 mg/l) for the root, callus and leaf proliferation from the leaf cutting slice. The highest number (6.75) of root proliferation was observed in the concentration of 2.0 mg/l IBA+0.25 mg/l BAP combination. The callus initiation was found in the concentration of IBA 1.0-3.5 mg/l+BAP 1.0-2.0 mg/l. However, the highest callus weight was observed at the concentration of IBA 1.5 mg/l+BAP 1.0 mg/l combination than other combination of concentrations. Positively leaf initiation and formation was better in the concentration of IBA 1-3.5 mg/l+BAP 1.0-2.0 mg/l combination. In addition, the 2,2-diphenyl-2-picrylhydarzyl (DPPH) free radical scavenging potential was higher (70.1%) in leaves extract than in callus extracts (46.3%) at the concentration of 10 mg/ml though both extracts had lower DPPH free radical scavenging activity compared to the positive control, vitamin C and BHT. Theresults conclude that the optimum concentration was IBA 1.5 mg/l+BAP 1.0 mg/l combination to produce callus cell proliferation and concentration of 2.0 mg/l IBA+0.25 mg/l BAP combination was the optimum for root proliferation of broccoli in vitro.
The study was conducted to evaluate the root, shoot and leaf callus cell regeneration and its biochemical properties like antioxidant, carbohydrate, pigment and mineral content from broccoli root, shoot and leaf cutting in vitro. An in vitro factorial experiment was carried out based on a Completely Randomized Design (CRD) with 5 replicates in tissue culture applying different IBA (0.25, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0 and 3.5 mg/l) and BAP (1 mg/l) concentrations using broccoli root tip and leaf cutting. The results showed that a higher callus weight was found in the cultured leaf cutting than in root tip cutting in the concentration of 1.0, 1.5 & 2.0 mg/l IBA + 1.0 mg/l BAP combination. The highest callus weight was found in the cultured leaf cutting than root tips cutting at the concentration of 1.5mg/l IBA+1.0 mg/l BAP. Furthermore, the highest inverted sugar and glucose, chlorophyll and nutrient content (K+, NO3- & Ca++), total phenol, flavonoid and total antioxidant were found in the concentration of 1.5mg/l IBA+1.0 mg/l BAP combination in both broccoli leaf and root cutting. The results seemed that it was best to use the combination of the IBA and BAP in the concentration of 1.0-2.0 mg/l and 1mg/l to regenerate root, leaf and callus cell proliferation of broccoli from the root tip and leaf cutting.