Study On Rapid Propagation And Bioreactor Culture Of Dendrobium Officinale

Dendrobium officinale is one of the most valuable Dendrobium species, and used in traditional Chinese medicine. Consumed extensively and in destructed habitat in recent years, the herb is in danger of extinction in nature. In order to realize its sustaninable utilization by mass propagation of protocorm-like bodies(PLBS) in bioreactor,these experiment compared the advantage-disadvantage of solid culture and liquid suspension culture,PLBS was used as material,the best culture condition for PLBS growth, multiplication and polysaccharides synthesize were investigated systemicly . The paper provided theory basis for PLBS tissue culture in mass propagation. The main results were showed as follows:1.Among the MS,B5,N6,1/2MS,1/4MS 5 media tested,1/2MS was suitable for the induction of the protocorm like bodies from the immature shoots, and MS medium was suitable for the subsequent subculture.2. To induction the PLBs by NAA, KT and 6-BA by orthogonal experiment, the result showed 0.1mg/L KT+2mg/L 6-BA+0.5mg/L NAA was optimal induction concentration. 1/2MS+1.0mg/L NAA+3% sucrose+5% coconut milk was the best medium for PLBs multiplication, and need some lights. 2mg/L BA +0.5mg/L NAA was suitable combination for the differentiation of calli. 0.5mg/L IBA +0.1mg/L NAA was the best suitable combination for the tube seedling growing and rooting.3. The results showed that the 1/2MS medium with 1.0mg/L NAA,50% coconut milk, and 3% sucrose was the most propitious to the proliferation of the PLBs. Inoculation with 40g PLBs in 1L liquid medium and pH 6.0 were the most suitable density for PLBs proliferations .4.Inner controllable circumstance of proliferation culture of Dendrobium officinale was adjusted using 500ml air lift bioreactor which was of column shape,the results showed the 1/2MS medium with 1.0mg/L NAA,5% coconut milk,3% sucrose , pH 6.0 and with 40g PLBs in 1L liquid medium was the most propitious to the proliferation and accumulation of polysaccharides of the PLBs by bioreactor culture; it was good for proliferation of Dendrobium officinale in bioreactor to use sparger with small pores of 15μm and afflux air volume of 0.1 vvm at the bottom.