Effect of Chlorine Dioxide on In Vitro Contaminated Plant and Plant Nursery by Azolla-Mixed Plant Material of Curcuma comosa

Abstract

Curcuma comosa is a herbaceous perennial plant belonging to the family Zingiberaceae. Rhizomes of this plant have been used in Thai traditional medicine as an anti-inflammatory agent particularly for the treatment of female postpartum uterine inflammation. Currently, conventional propagation of C. comosa is too low multiplication rate. Using tissue culture technique is a suitable protocol for mass propagation in a short time. However, sterilization of explant was very important sterilization by chemical substance is a potent method. Chlorine dioxide could reduce microorganism in culture medium and explant and increased efficiency in micropropagation by bioreactor system. The purposes of this research were to reduce microorganism in culture medium by Chlorine dioxide; and to realize the use of azolla as culture media for hardening of  C. comosa. Sterilized culture medium with ClO₂ promoted shoot formation and removed fungal contamination of C. comosa culture. ClO₂ at 30 mg/L (10 mg/L, 3 times) gave best result in contamination (0%) and number of shoot (6.86±0.55 shoots). In the experiment of plant survival rate, Azolla for culture medium composed of sand: coconut coir: Azolla at ratio of 2.0: 1.0: 0.1 by volume gave the highest survival rate at 100.00 percent at 30 days of culture. These results would be useful for future study of other Zingiberaceae.