| In this study rosemary cell suspension culture was used as material,as well as carbon source and nitrogen source were added to basic medium for investigating the effect on cell gro.Arth and secondary metabolites accumulation and antioxidant enzyme activity.Meanwhile,salicylic acid and methyl jasmonate were added to basal medium for investigating their effects on the cell growth,secondary metabolites accumulation,DPPH activity and relevant enzymes activity involved in secondary metabolites synthesis(phenylalanine ammonia enzyme PAL and tyrosine aminotransferase TAT).The aim of this study was to obtain the optimum growth condition of rosemary cell suspension culture.Ultimately it will be significant to supply technical parameters or regulating the production of secondary metabolites by means of cell suspension culture.Main conclusions were as follow:1.The effect of carbon source on physiological and biochemical characteristics of rosemary cell suspension cultureWe concluded that 30g/L sucrose,70 g/L maltose and 40g/L glucose were benefit to the growth of rosemary cell suspension culture.70g/L sucrose,70g/L maltose and 70g/L glucose were benefit to the acid accumulation of rosemary cell suspension culture.lOg/L sucrose,20g/L maltose and lOg/L glucose were benefit to the carnosic acid accumulation of rosemary cell suspension culture.The combination of growth and secondary metabolite content point of view,30g/L sucrose,30g/L maltose,and 40g/L glucose The combination of growth and secondary metabolite content point of view,30g/L sucrose,30g/L maltose,and 40g/L glucose were benefit to rosemary cell suspension culture.Through comparison analysis,sucrose of 30 g/L had the highest growth rate and content of secondary metabolites in the three kinds of sugar source.It is concluded that high concentration of sucrose processing would obviously enhance the content of H2O2 and MDA in cell suspension culture,and meanwhile,the activity of antioxidant enzymes SOD,POD and CAT were also enhanced significantly by the determination of antioxidant enzyme system of sucrose.It is indicated that a high concentration of sucrose would produce osmotic stress,and it was not conducive to growth of rosemary cell suspension culture,but it was conducive to the accumulation of secondary metabolites.Eventually,we concluded that sucrose of 30 g/L was most conducive to rosemary suspension cell culture according to the rosemary suspension cells growth rate and rosemary acid content.2.The effect of nitrogen source on physiological and biochemical characteristics of rosemary cell suspension cultureUnder the treatment of different nitrogen sources,the growth of rosemary cell suspension culture and accumulation of secondary metabolites had optimum proportion when ammonium/nitrate nitrogen was 2:3.,rosemary and carnosic acid content of the acid also reached the maximum.The rosemary cell suspension culture had the highest growth rate,and rosemary acid content increased 1.57 times compare to the normal rosemary leaves,carnosic acid nitrogen content is about 13 times the rests.Under the optimum proportion,the MDA concern and antioxidant enzyme activity were the highest.It is indicated that membrane peroxidation happened,and antioxidant enzyme system showed strong activity,then the accumulation of reactive oxygen species in the cell was controlled effectively,and promoted plant growth and synthesis of secondary metabolites.3.The effect of salicylic acid on growth,secondary metabolites accumulation,synthetase and antioxidant activity in rosemary cell suspension cultureIn the processing of different salicylic acid concentrations,the cell suspension culture growth rate could be effectively improved under salicylic acid concentration of 6.25 mg/1 and incubation time 72 h.When the concentration of salicylic acid was 25 mg/1,it contributed to the rosemary acid and camosic acid synthesis at incubation time 24 h and 96 h,respectively.To some extent,the inducing effect of salicylic acid was a significant positive correlation with the activity of PAL and TAT.However,adding the salicylic acid could not improve antioxidant activity of cell suspension culture.25 mg/1 salicylic acid was optimum incubation condition for the growth and secondary metabolites accumulation in rosemary cell suspension culture.4 The effect of methyl jasmonate on growth,secondary metabolites accumulation,synthetase and antioxidant activity in rosemary cell suspension cultureIn the processing of different methyl jasmonate concentrations,the cell suspension culture growth rate could be effectively improved under salicylic acid concentration of 100μmol/l and incubation time 96 h;The content of secondary metabolites in cell suspension culture would be improved significantly at the appropriate methyl jasmonate concentration(100-150μmol/l),The ability of increasing rosemary acid and camosic acid accumulation was significantly positive correlated with the activity of PAL after adding methyl jasmonate into rosemary cell suspension culture.To some extent,adding methyl jasmonate promoted antioxidant activity of the cell suspension culture.In conclusion,it’s the most conducive to rosemary cell suspension culture grow and synthesis secondary metabolites at 100μmol/L methyl jasmonate. |
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