Establishment of plant regeneration protocol through callus induction of sugarcane (Saccharum officinarum l.) var. China using leaf-sheath

Establishment of a plant regeneration system through direct organogenesis or via callus is also a prerequisite to further in vitro genetic manipulation of the cultivar through somaclonal variation, in vitro mutagenesis and genetic transformation. Production of sufficient numbers of plants of a unique genotype is possible using in vitro culture system. In this study, the effect of various concentrations and combinations of plant growth regulators for in vitro regeneration of sugarcane via callus was described using leaf-sheath as explants. Thus, a reproducible protocol for induction of callus and regeneration into plantlets were developed in sugarcane var. china using leaf-sheath as explants. Potential callus induction was observed on MS medium supplemented with 4.5 mg/l 2,4-D, in which 80% explants induced callus after culture initiation of 90 days. Shoot formation from the callus was optimum on MS + 1.0 mg/l GA₃ + 0.5 mg/l Kin., in which 70% of the cultured calli formed shoots. The average number of shoot formed per culture callus was 27.0 ± 2.50 and the average shoot length of 6.5 ± 0.50 cm were achieved in this medium after culture of 60 days. Regenerated shoots rooted well when they were transferred into half strength of MS + 2.50 mg/l NAA, in which 90% shoots rooted within 30 days of culture. The average number of root produced per shoot was 18.0 ± 2.25 and the average root length of 7.50 ± 1.20 cm were observed in this medium. Ninety percent of the in vitro raised plantlets were survived in the natural environment.