Establishment of Sterilization Method for Emergent Quality of Sugarcane (Saccharum officinarum L.) In An Efficient Micropropagation System.

The present investigation focuses on development of economical, reliable and effective sterilization protocol for micropropagation of sugarcane CoVSI 9805 with isolation and identification of contaminating microbes. Surface sterilization was carried out using different formulations of mercuric chloride, Bavistin (TM) and Streptomycin. Aseptic conditions in plant shoot generation medium were studied with use of different concentration of plant preservative mixture (TM) (PPM), sodium hypochlorite (NaOCl), chitosan (I and II). The isolated and identified contaminating microbes comprised of four fungi viz. Alternaria, Aspergillus, Penicillium Fusarium spp. and five bacteria viz. E. coli, Bacillus subtilis, Staphylococcus aureus, Agrobacterium tumefaciens and Pseudomonas aeruginosa respectively. The occurrence of bacterial contaminants in in vitro culture was higher than the fungal. After surface sterilization, maximum proliferation (55.33 +/- 1.45) with reduced necrosis (20 +/- 1.15) was obtained with combination of Bavistin (0.1%) + streptomycin (0.1%) at 25 +/- 1 degrees C for 15 min. The sterilizing agents decontaminated bud surface and enhanced proliferation but single sterilant alone failed to control contamination. An increased response of axillary bud in vitro culture was achieved with PPM (0.5%), NaOCl (0.01%), chitosan I (0.01%) and chitosan II (0.03%). The improved control contamination strategy will result in mass multiplication of sugarcane by tissue culture and augment the proliferation to compete agriculture demands.