Optimization of Protocol for Callus Formation and Shoot Development in Sugarcane (Saccharum officinarum L.) Cultivar CPF-248

Abstract

Sugarcane production face numerous difficulties, including infestations of insects and diseases, particularly red rot, a lack of appropriate seed supply, a lack of industrial support, etc. Using a process called plant tissue culture, huge amounts of authentically grown, disease-free plant material may be produced quickly. Plant tissue culture can also be used to quickly reproduce recently released varieties with crucial agronomic characteristics. For this purpose, sugarcane callus culture was collected from the inner soft leaf sheath to increase genetic diversity. Ten different concentrations viz., 1.5, 1.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5 and 6.0 mgL-1 of 2, 4-dichlorophenoxy acetic acid in MS medium were used for callus development, along with 0 mgL-1 control was used. Different combinations and concentrations of BAP+ Kin (1.0+1.0, 1.0+1.5, 1.0+0.5, 1.0+1.0, 1.5+1.0 and 1.0+1.5 mgL-1) were used for regeneration of shoots and auxin. IBA with 6 different concentrations (0. 0.3, 0.5, 0.7, 1.5 and 1.8 mgL-1) were used for rooting of the shoots. Among all the growth regulators, 2, 4-D at 3.0 mgL-1 demonstrated the greatest auxin for callus development with BAP. The 6-12 mm meristem was grown in MS medium with NAA. It was observed that at concentrations viz., 3.5mgL-1 and 4.0mgL-1 of 2, 4-D, maximum callus (79.0-84.5 %) was developed with 4-5 mm in size. Maximum root growth and length (3.7 mm) were reported at 1.0 mgL-1 IBA and 1.0 mgL-1 Kinetin and 1.5 mgL 1 BAP in MS medium. It was concluded that auxin concentration is best for all sugarcane in vitro callus culture, which will assist researchers in future work.