Biological effect of irradiated chitosan on plants in vitro

For degradation of chitosan, chitosan with an 80% degree of deacetylation and a weight-average molecular mass (Mw) of approx. 48 kDa was irradiated with gamma-rays at doses up to 200 kGy in a 10% (w/v) solution. The Mw of chitosan was reduced from 48 to 9.1 kDa by irradiation. The characteristics of irradiated chitosan were analysed by using Fourier-transform IR spectroscopy and an elemental analyser. The amino group was found to be stable, whereas the C-O-C group decreased with increase in the dose. The product of chitosan irradiated at 100 kGy with an Mw of approx. 16 kDa showed the strongest growth promotion effect on plants in vitro. For shoot culture, supplementation with irradiated chitosan increased the fresh biomass of shoot clusters (7.2-17.0%) as well as the shoot multiplication rate (17.9-69.0%) for Chrysanthemum morifolium (florist's chrysanthemum), Limonium latifolium (limonium or sea-lavender), Eustoma grandiflorum (lisianthus, tulip gentian or Texas bluebell) and Fragaria ananassa (modern garden strawberry). The optimum concentrations of irradiated chitosan were found to be approx. 70-100 mg/l for chrysanthemum, 50-100 mg/l for lisianthus and 30-100 mg/l for limonium. For the plantlet culture, the optimum concentrations were found to be approx. 100 mg/l for chrysanthemum, 30 mg/l for lisianthus, 40 mg/l for limonium and 50 mg/l for strawberry. Supplementation with optimum concentrations of irradiated chitosan resulted in a significant increase in the fresh biomass (68.1% for chrysanthemum, 48.5% for lisianthus, 53.6% for limonium and 26.4% for strawberry), shoot height (19.4% for chrysanthemum, 16.5% for lisianthus, 33.9% for limonium and 25.9% for strawberry) and root length (40.6% for chrysanthemum, 66.9% for lisianthus, 23.4% for limonium and 22.6% for strawberry). In addition, treatment with irradiated chitosan enhanced the activity of chitosanase in treated plants and also improved the survival ratio and growth of the transferred plantlets acclimatized for 10-30 days under greenhouse conditions.