Study On Optimization Of Culture Condition Of Perilla Frutescens Suspension Cell

Take the Perilla frutescens plant as material to built and optimize the suspension system of Perilla frutescens cell. The sterilization of plant, the induction of leaves, the establishment of suspension system and optimize the contidions of suspension cell were researched in this paper. The restults as follows:First, the stems, leaves, and cotyledon of Perilla frutescens were treated as explants with0.1%HgCl2and2%NaClO according to different time. The result showed that choose the leaves as the explant to induce callus according the induction ratio and infection ratio according to different time. The result showed that the optimal sterilization method for the leaves was75%aleohol for30S and0.1%HgCl2for7min. On this condition, the induction ratio is80.7%and the infection ratio is5%.Second, the effects of different concentrations of the hormone combination on inducing callus in B5medium and MS medium were studied by orthogonal test. Considering the induction ratio and the growth of callus, the optimal medium for callus induction is B5+2,4-D1.0mg/L+KT0.4mg/L+6-BA1.0mg/L+NAA1.5mg/L which could achieve the highest induction rate.Third, to survey and evaluate the contents of suspension cell and the optimal medium is B5medium+pH6.0+sucrose4.0%+nositol150mg/L+6-BA2.0mg/L+2,4-D2.0mg/L+KT0.4mg/L+NAA1.5mg/L by Single-Factor experiment and orthogonal test.Forth, the production of rosmarinic acid was estimated by HPLC, the results showed that, the rosmarinic acid production of stems is0.72%, the callus contained more rosmarinic acid than leaves of Perilla frutescens,3.85%and0.25%dry weight, respectively. And there are15.8mg/g～26.1mg/g in suspension-cultured cell(dry weight).