Vol. 12, Issue 9 (2023)

Author(s):
Muzamil Yousuf, Dr. Gurdev Chand and Dr. BK Sinha

Abstract:
An in vitro efficient complete protocol has been developed for plant regeneration through callus induction in the medicinal plant Broccoli (Brassica oleracea var. italica) using leaf as ex-plant. Tissue culture is a new approach for production of vigorous and disease free plants with many advantages over conventional methods. Broccoli (Brassica oleracea var. italica) is a valuable source of antioxidants and secondary metabolites. This study is a first extensive report about the effect of different serialization protocols and plant growth regulators (PGRs), and nanoparticles viz. Chitosan (50nm) on different explants disinfection efficiency, callus induction and accumulation in callus cultures of Broccoli. In addition, different sterilization agents were evaluated for disinfection of leaf explants, nodal explant and seed explant. The results revealed that out of 12 treatments on surface sterilization of nodal segment explants, the best treatment was T₁₀ consisting of 1.0% (w/v) bavistin for 10 minutes followed by 0.50% (v/v) sodium hypochlorite for 5 minutes duration and 0.25% mercuric chloride (HgCl2) for 1 minute. Similarly for seed explants that treatment T₁₁ comprising of 1% (w/v) bavistin for 10 minutes duration followed by 0.50% (v/v) sodium hypochlorite of 15 minutes and mercuric chloride 1% for 1.50 minutes gave 85.51% uncontaminated cultures in case of seed segment explants, which is the highest as compared to all other treatments. The results presented for leaf explants showed that treatment T₇ comprising of 1% (w/v) bavistin for 5 minutes duration followed by 0.50% (v/v) sodium hypochlorite of 3 minutes and mercuric chloride 0.1% for 0.30 minutes gave 88.75% uncontaminated cultures in case of leaf segment explants, which is the highest as compared to all other treatments. Sterilization protocols adversely influenced the explant viability and callus induction and growth. On culture media (cl7) supplemented with 1.5 (mg/l) 2,4-D and 0.75 (mg/l) Chitosan showed the maximum callus induction frequency (CIF) (90.27%),: The results indicated the positive effect of low dosage of nano elicitors on callus induction and explant viability. Furthermore, the callus induction is tightly dependent to the presence of auxin and cytokinin, which stimulate both cell division and cell elongation.