Establishment Of Cephalotaxus Cell Suspension Culture System For Cephalotaxus Mannii

Using the callus from tender leaf of C. mannii for fluid suspension cultivation,this paper mainly focus on the establishment of C. Mannii cell suspension culture system,the kinetic analysis of suspension cell growth, secondary metabolites synthesis and nutrient consumption, the effects of different kinds of Osmotica regulator on the cell growth and flavonoids accumulation were studied.The results were mainly shown as follows:1?The factors influencing the establishment of C. mannii cell suspension culture system, such as culture medium,Vitamin,sucrose concentration,pH,exogenous hormones,inoculum amount,were investigated.The alkaloid content in cell suspension culture system of C. mannii was determined by HPLC.The results showed that the optimal medium for the cell suspension culture was on the MS medium supplemented with4mg/L NAA and O.lmg/L KT under dark, sucrose concentration was35g/L, inoculation fresh weight was6%, VB1concentration was lOmg/L and initial pH5.5-7.0;It is useful for cell growth and harringtonine formatio,If justing pH to6.0every fifth day in the experiment.2?The cells growth basically accorded with the " S "growth curve,the cell growth cycle is30days,growth lag was0-10days,logarithmic growth phase was10-25days,Stable growth was25-30days,products slowly decresed on the0-15days,Mainly for primary metabolism,cell division and proliferation,and the greatest products obtained on day25-30days;the sugar consumption accorded with inverted " S " curve,Its consumption process coincide with each phase of cell growth;Ammonium salt consumption in cultured before10days significantly faster than the nitrates cells,then ammonium concentration decreased slowly while nitrate consumed faster,phosphate consumed quickly in growth lag;viability sharply increased at first and gradually decreased in later.3?The effects of different kinds of osmotic regulator on the cell growth and flavonoids accumulation were studied.The results showed that it is useful for cell growth and harringtonine formatio when1%mannitol added in the logarithmic growth phase.