Improving the micropropagation efficiency of hybrid Dendrobium orchids with chitosan

The appropriate chitosan types and concentrations for enhancing the in vitro micropropagation of Dendrobium ‘Eiskul’ were studied using 70, 80 and 90% N-deacetylated polymeric (P-70, P-80 and P-90) and oligomeric (O-70, O-80 and O-90) forms of crab (Portunus pelagicus) chitosan. For the initial protocorm-like body (PLB) multiplication, the application of 10 mg/L of P-70 or 20 mg/L of P-90 was optimal, although 10 mg/L of P-80 and O-70 were also effective, and attained maximal PLB replication rates without increasing the detectable levels of somaclonal variation. However, during PLB-shoot induction, 10 or 20 mg/L of O-80 was the most appropriate chitosan and also induced further PLB formation. For plantlet regeneration, the addition of 10 mg/L of O-80 or P-80 gave the best quantity and quality, respectively, of plantlets. Finally, 20 mg/L of P-70 chitosan as a supplement during exflasking enhanced both the survival rate and the growth of the plantlets at one month after exflasking. Together, these data reveal a potentially beneficial and applicable protocol for commercial orchid micropropagation.