| As one of versatile pharmaceutical intermediates, rosmarinic acid has many functions including antioxidant, antibacterial and anti-inflammatory, anti-virus and immunomodulatory. Perilla frutescens is rich in rosmarinic acid. This study was conducted to identify initially the suitable conditions for callus growth and rosmarinic acid biosynthesis by plant tissue culture techniques, and establish a perilla suspension cultures system with a high yield of rosmarinic acid. The results were as follows:1.Influence of plant growth regulators on callus culture in perilla was studied, and the results showed that a single hormone was not suitable for callus induction but combination of BA and NAA can induce more Perilla calllus. The optimum medium for callus induction was MS+BA3.0mg/L+NAA0.3mg/L.2.The optimal culture conditions were determined for callus induction culture. Perilla hypocotyl explants inoculated on solid medium were cultured firstly in the dark for 10 days, then transferred to train under 10 h/d light condition at 24℃.The time of subculture circle was 20d/Generation. After 4 times subculturing, callus can be put into liquid medium for suspension culture.3.In suspension culture system, the optimum condition for perilla callus cell growth and rosmarinic acid biosynthesis was as follows:initial inoculum 0.9g/flask, culture period 7 days, and shaking speed 110r/min. The optimal concentration of sucrose was 30g/L for perilla cell growth and for rosmarinic acid biosynthesis was 40g/L. Phenylalanine had significant effect for inducting cell growth and promoting rosmarinic acid biosynthesis. Dry weight of cultured cell reached the maximum growth amount when phenylalanine concentration was 0.1 g/L, while the highest percentage of rosmarinic acid in total dry weight reached 2.283% when it was 0.15g/L.
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