| Panax ginseng C. A. Meyer, Panax quinquefolium L. and Glycyrrhiza uralensisFisch. are widely used herb. In this study, adventitious root of P. ginseng, cells of P.quinquefolium and G. uralensis have been established. We also conducted studies onbioreactor culture.Callus of ginseng was induced by roots of P. ginseng （5-year-old）. Adventitiousroots were initiated from callus which were inoculated onto MS solid mediacontaining5.0mg·L-1IBA. Growth rate of50-fold in5L balloon-type bubblebioreactor （BTBB） was obtained after40days of inoculation. The maximum totalsaponin was achieved on day30. Rg1, Re, Ro, Malonyl-Rb1, Rb1, Rc, Rb2and Rdwere identified from ginseng adventitious root. Ginseng adventitious root culturegrew faster and had a greater capability of ginsenoside production. With24h of10mg·L-1MJ elicitation, level of total saponins in ginseng adventitious root increasedmuch higher than that observed in the control, especially Rb group, which wererelated to the expression of SE, DS and P450. DPPH inhibition of ginsenoside andpolysaccharide were higher in adventitious roots than in native roots, with60mg L-1ginsenoside of ginseng adventitious roots, the DPPH inhibition was96.03%.Callus of P. quinquefolium was induced by roots of P. quinquefolium （5-year-old）.Cells were initiated from callus which were inoculated onto MS solid mediacontaining2mg·L-12,4-D,0.25mg·L-1KT. In a bioreactor, the dry cell weight, thecontents of ginsenosides and polysaccharide reached the maximum peaksimultaneously on about21days and the results showed that cell growth andmetabolites synthesis related to nutrients consumption. LH at100mg L-1with methyljasmonate （MJ） at2mg L-1synergistically stimulated ginsenoside accumulation in P.quinquefolium cells compared with100mg L-1LH. Using a fed-batch cultivationstrategy, polysaccharide production was enhanced to1.608g L1, which was1.96-foldgreater than with batch cultivation. Two-stage cultivation caused a significant increasein total saponins yield (31.52mg·L-1) and polysaccharide yield (1.72g·L-1) cellcultures. This value was increased by4.34-fold and2.1-fold compared to the batchcultivation, respectively. Rg1, Re, Rf and Rb1were identified from P. quinquefoliumcells. The higher polysaccharide content is an obvious advantage of P. quinquefolium cells. However, the ginsenoside contents in the cell cultures are still much lower thanthat in field cultivated. DPPH inhibition of ginsenoside in P. quinquefolium cells washigher than that in native root. DPPH inhibition of polysaccharide both in native P.quinquefolium and cells are lower.Callus of G. uralensis was induced by hypocotyls of G. uralensis. Cells wereinitiated from callus which were inoculated onto MS solid media containing1.0mg·L-12,4-D,1.0mg·L-1NAA,0.2mg·L-16-BA. The maximum dry weight,triterpenoids and flavonoids were achieved on day20,10and15, respectively in10LBTBB. Maximum growth rate of10.86-fold in10L BTBB were obtained after18days of inoculation. Liquiritin, licorice glycoside B, licorice saponin J2, glycyrrhizicacid and licorice saponin B2were identified from G. uralensis cells. Triterpenoids in G.uralensis cells are much less than that in native licorice. However, flavonoids in cellsare similar to the native licorice. Content of polysaccharide in cells （11.7%） is higherthan that in native. About93.98%and100%DPPH inhibition occurred with500mgL-1flavonoids of native licorice and cells. DPPH inhibition of polysaccharide in nativelicorice and cells are lower.These results provide a theoretical reference for an efficient production of activemetabolites of above three kinds of plants. |
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