Cell suspension culture of Dendrobium huoshanenes and its optimal conditions / 中草药

ABSTRACT

Objective: Single-cell of Dendrobium huoshanenes was cultured by using cell suspension culture for solving the problem of resource shortage of medicinal plant D. huoshanenes. Methods: The single-cell of D. huoshanenes was obtained by different preparing ways, effects of various factors on cell growth were studied and the optimal growing conditions of cell suspension culture were investigated by orthogonal design. Results: Through the orthogonal test, the results indicated that the optimum conditions for suspension culture of D. huoshanenes cell were as follow is that the concentration of sucrose was 35 g/L, NH4NO3 was 1.3 g/L, pH value was 5.6, the hormone IAA was 0.4 mg/L , 6-BA was 1.0 mg/L, the best culture time was 14-18 d. Under optimal culture conditions, the concnentration of single cells in this test was 9.42 X 105 cell/mL. Conclusion: After four successive subculture, cellular morphology of D. huoshanenes shows circular shape or elliptic shape and the chip of cell is decreased. The higher concentration of D. huoshanenes cell could be obtained by using the optimal growing conditions of cell suspension culture.