Establishment Of Regeneration System And Researches On Biological Activity Of Volatile Oil Of Echinacea Purpurea L

Echinacea purpurea L., also named as Echinacea, is a kind of perennial herb, belonging to composite family. It is a kind of medicinal plants which has great development value for its prominent value in immune function, anti-bacterial effects, anti-inflammatory effects and antiviral effects. The regeneration system of Echinacea purpurea L was established in this study by the technology of tissue culture. The chemical constituents of different parts of volatile oil of Echinacea purpurea L were also analyzed by GC-MS. The antioxidant effectbacteriostasic activity in vitro and inhibition of tyrosinase activity of the volatile oil from Echinacea purpurea L were researched to offer theoretic basis and technological support for the plantation and exploitation of E. purpurea. The research results are as follows:1、We build an effective and quick isolated regeneration system of Echinacea purpurea L. The experiments studied the optimum soaking concentration of gibberellin and the best treatment of the seed sterilization on seed germination, then the best way among them was chosed. Using leaves as sterile sprouts, the effects of different phytohormones as well as their combinations to callus induction, adventitious bud induction and proliferation and differentiation of adventive roots were studied, and the most suitable medium for the tissue culture of Echinacea purpurea L was chose. The outcomes were as follows:the seed germination was promoted with the gibberellin soaking of400mg/L. The best sterilization method for seed ofhinacea purea L. was75%alcohol for30seconds and0.1%mercuric chloride for10minutes. The optimum condition on callus induction was to culture in dark for one week, then culture in light and the optimum combination of hormone was MS medium+6-BA1.0mg/L+NAA1.5mg/L+2,4-D1.0mg/L. The best combination of hormone on induction of adventitious buds was MS medium+6-BA1.0mg/L. The suitable hormone concentrations on proliferation of adventitious buds was6-BA2.0mg/L+NAA0.1mg/L. MS medium+IBA0.5mg/L was the best medium for the differentiation of adventitious roots; The most suitable matrix of plantlets transplanting was vermiculite:perlite:peat (1:1:1).2、The chemical constituents of different parts of volatile oil of Echinacea purpurea L. was analysed by GC-MS. The result showed:36components were isolated and identified from the volatile oil.25components belong to terpenoid, the content of which was91.39%, and all were sesquiterpenes.1component (0.65%) belongs to aromatic compound,10 components belong to aliphatic compound (7.53%). The main components of the flower were beta-ylangene (15.8%), cubebene (8.13%), Germacrene D (20.84%).30components were isolated and identified from the volatile oil of the root.10components belong to aliphatic compound, accounting for65.84%of the total volatile oil.19components belong to terpenoid, accounting for33.49%,2,4-decadienal (E, E)-was monoterpene and the rest were sesquiterpenes.1component (0.68%) belongs to aromatic compound. The main components of the root were1,11-dodecadiene (12.49%), n-hexadecanoic acid (36.13%),9,12-octadecadienoic acid (Z,Z)-(10.44%).14components were the same between the flower and the root:alfa-copaene,(-)-beta-elemene, alpha-bergamotene,1,11-dodecadiene, cubedol,(+)-d-cadinene, caryophyllene oxid, alloaromadendrene, alpha-cadinol, naphthalene,1,6-dimethyl-4-(1-methylethyl)-,n-hexadecanoicacid,9,12-octadecadienoicaci d (Z,Z)-, tetracosane, eicosane.3、The antioxidant effects of volatile oil ofhinacea purpurea on rapeseed oil and lard were studied. We used peroxide value, acid value and TBA value as indexes, rapeseed oil and lard as mediums, compared with the synthetic antioxidants BHT and BHA. The results showed that within the scope of the study, the antioxidant effects of volatile oil on rapeseed oil and lard were existed and enhanced with the increasing amout. The antioxidant effect was obvious on rapeseed oil with the addition of2g/kg volatile oil and the influence was almost equivalent to0.2g/kg BHT and BHA. The antioxidant effect of the volatile oil on lard was less than rapeseed oil.4、The inhibition of volatile oil on the activity of tyrosinase was studied. The inhibition effect was determined by spectrophotometer, using L-tyrosine and catechol as the substrate, and the influence was compared with the arbutin. The results showed that the inhibition effects of the volatile oil on monophenolase and disphenolase activities were existed and enhanced with the increasing amout. The inhibition effects of the volatile oil were both stronger than arbutin.5、Bacteriostasic activity in vitro of the volatile oil was studied. The inhibition effects on acteria and mould were studied by cup method, the MIC of acteria and mould were also studied by fold dilution method, taking potassium sorbate as control group. Results showed that the volatile oil had inhibition effect on bacteria and the strength was Slaphylococcus aureus> E. coli>Bacillus subtilis>Micrococcus tetragenus. The MIC for Staphylococcus aureus, E. coli, Bacillus subtilis and Micrococcus tetragenus were0.625%、1.25%、2.5%and2.5%, respectively. The inhibiton effect on mould was not clear.