ObjectiveThe proliferation of a great number of active embryogenic callus can provide sufficient materials for somatic embryogenesis and plant regeneration of Pinus tabuliformis. However, the yield of flask suspension proliferation is limited and easy acidification necrosis. The establishment of airlift bioreactor system for embryogenic callus multiplication of P. tabuliformis can promote the proliferation of embryogenic callus of P. tabuliformis.
MethodIn this study, P. tabuliformis callus was used as material, and L9 (34) orthogonal design was used to investigate the effects of three factors on callus proliferation in bioreactor, including inoculation volume, ratio of old and new media and hormone concentration. Then the suspension culture in conical flask was compared with that in conical flask under the same conditions.
ResultThe highest embryogenic callus proliferation rate, 216.18% was obtained using 10 g embryogenic callus, 0.2 mg/L 2,4-D and 20% old culture medium inoculated in 100 mL liquid medium in the ALB.
ConclusionCompared with conical flask suspension culture, the growth rate of embryogenic callus in ALB system was 2.15 times faster in a week. Microscopic observation shows that the proliferated embryogenic callus is stable and high quality. This study provides technical support for large-scale propagation of P. tabuliformis based on somatic embryo system.