Callus Cell Proliferation and Explants Regeneration Using Broccoli Shoot Tip in vitro Culture: Biochemical and Antioxidant Properties
A. B. M. Sharif Hossain *
Department of Biology, Program of Biotechnology, Faculty of Science, University of Hail, KSA and Program of Biotechnology, ISB, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia.
Imdadul Haq
Program of Biotechnology, ISB, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia
Mohammed Saad Aleissa
Department of Biology, Faculty of Science, AI-Imam Muhammad Ibn Saud Islamic University, KSA
Nasir Adam Ibrahim
Department of Biology, Program of Biotechnology, Faculty of Science, University of Hail, KSA
Kamaludin Bin Rashid
Program of Biotechnology, ISB, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia
*Author to whom correspondence should be addressed.
Abstract
Significance of the Study: Nowadays cell or tissue culture has been successfully performed using different types of species like fruit, vegetable, ornamental and forest plant. Millions of explants can be produced by tissue or cell culture per year in any 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.
Aims: The study was carried out to investigate the root, callus, shoot and leaf proliferation from the shoot tip in vitro culture using different IBA and BAP concentration.
Methodology: Shoot tip slice was used to culture in the MS (Murashige and Skoog) media. Different IBA and BAP (0, 0.25, 0.50, 1.0, 1.50, 2.0 2.5, 3.0 and 3.5 mg/l) concentrations in combination with MS media were used.
Results: The highest number (7.4) of shoot proliferation was observed in the concentration of IBA 0.25+BAP 2.5 mg/l combination. The maximum root proliferation was found in the concentration of IBA 0.25+ BAP 1.5. However, callus formation was observed better at the concentration of BAP 1-3.5 and IBA 1-3.5 mg/l combination than other combination of concentrations. 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. Besides, both extracts had lower DPPH free radical scavenging activity compared to the positive control, vitamin C and BHT.
Conclusion: The present results conclude that it is better to use the combination of the IBA and BAP concentration to produce root, shoot, leaf and callus cell proliferation in broccoli.
Keywords: Broccoli, Brassica oleracea, callus cell, in vitro culture, BAP, IBA, DPPH