The aquatic habit of lotus makes obtaining aseptic cultures from field-growing plants
very difficult, therefore, surface sterilization was carried out using a sequence of rinses in
sterilizing agents. Buds from rhizomes of field-grown plants of lotus cv. Satabankacha
were initially rinsed in 70% ethanol for 1 min. Explants were then transferred to 0.1% (w/
v) mercuric chloride (plus two drops of tween20) for 10 min followed by 30 min in 5% (w/
v) calcium hypochlorite (plus two drops of tween20) and then 10 min in 1% calcium
hypochlorite (plus two drops of tween20). Finally, the explants were rinsed three times in
sterile distilled water.