Use of Response Surface Methodology on Medium Components Optimization for Embryogenic Callus Induction in Sugarcane (Saccharum Sp.) Cv. Cp 72-2086

Abstract

The importance of sugarcane culture has in the agriculture and industrial market, demand the development of efficient in vitro propagation systems; in this sense embryogenic callus has a great impact in basic research and industrial applications, due to this, the development of protocols for embryogenic callus induction is of great importance. In this work, an optimized protocol for embryogenic callus induction was developed; to do this, the balance of the phytohormones 2,4-dichlorophenoxyacetic acid (2,4-D) and indole-3-butyric acid (IBA) in the culture media composition was optimized using response surface methodology (RSM) in order to maximize the percentage of samples with embryogenic callus (%SEC) and the weight gain on callus produced (ΔCW) in Saccharum sp. cv. CP72-2086. Best results in the %SEC were obtained as the 2,4-D concentration decreased; meanwhile, in the case of IBA the best results were obtained in the concentration range from 2.5 to 5.5 mg L-1. Regarding ΔCW, maximum values were obtained at 2,4-D concentrations in the range of 0.5 mg L-1 to 1.89 mg L-1 in combination with IBA in the range of 4.55 mg L-1 to 5.25 mg L-1. The best predicted response was obtained at a combination of 0.5 mg L-1 2,4-D + 3.53 mg L-1 IBA, from which the predicted values of the model were 100.0 of %SEC and 1.4323 g of ΔCW. The experimental values were very close to those predicted by RSM, which suggests that a computer-based experimental design it is very useful to optimize protocols for successful embryogenic callus induction in sugarcane.