Establishment Of Culture System Of Echinacean Purpurea Hairy Roots And Study On Secondary Metabolites

Echinacea purpurea,which belongs to Compositae familia,is a traditional North American perennial medicinal herb.It has many effective medicinal ingredients,such as polysaccharides,caffeoyl phenols,and alkylamides,so as to improve body immune system. Accordingly,it has become one of the most popular herbal products in North America and Europe as an immune promoter,and should have widely applied prospect in medicine. Echinacea purpurea,famous for its distinct pharmacological efficacy and commercial value,has been using to diminish inflammation,kill virus and bacterium,as well as to enhance people's immuno competence as a kind of herbal medicine.It has 8 kinds and several varieties,is the perennial herb,capitulum on the stem or an item,its chief feature is the receptacle is the cone, has the tubular flower and the ligulate flower,usually for rose color or purple.Antibacterial and the adjustment organism immunity function has the remarkable pharmacological action and the extremely high for medicinal purposes value.Hairy roots induced by Agrobacterium rhizogenes have the ability to grow rapidly,and it is easy for hairy roots to control and operate.They also have physiologic,biochemical and genetic stability,showing unique superiorities and large potential for production of secondary metabolites.In this paper,aculture system of E.purpurea hairy roots was established,effects of sound stimulation on growth of the induced hairy roots and accumulation of their secondary metabolites were studied.In this article,3 kind Agrobacterium rhizogenes of A4,R1601,1025 were used to induce the genetic transformation of E.purpurea,Establishment and identification of a system to transform hairy roots of E.purpurea.Hairy roots were induced from the wounded brim of the explants,30 days after inoculation of E.purpurea explants with A.rhizogenes of A4,R1601 or 1025.Hairy roots branched rapidly and grew upwards.The percentage of rooted leaf explants was 66.7%after A4 infection.At the same time,single-factor experiments for several influential factors were carried out to find the best condition for inducement of Echinacea purpurea hairy roots.As a result,the most appropriate A.rhizogenes was A4,the most appropriate infecting time was about 12min,the most appropriate OD600 of A4 suspension was about 0.8,the most appropriate coculture time was 2 days,the most appropriate proculture was 2 days,the most appropriate density of AS was 50umol/L,the most appropriate coculture temperature was about 24℃and the most appropriate pH of culture medium was about 6.0,under which the average inducing rate reached 54%.The induced roots were identified and confirmed to be transformed hairy roots by PCR analysis.Studies on the chemical constituents content determination of secondary metabolites in hairy roots of E.purpurea,such as polysaccharide and phenolic.The transformed hairy roots grew rapidly after 60 days on a MS medium.By analyzing extraction of hairy roots,it was showed that the content of polysaccharides and phenolic compounds were 15.54%and 2.491%.