Establishment Of Transform System In Eucommia Ulmoides Oliv. And Studies On Hairy Root's Microstructure And Its Secondary Metabolite

Eucommia ulmoides Oliv.is a very old and well-known drug in traditional Chinese medicine, which is the only arbor of Eucommiaceae.The high physiological activity of the root extracts is a consequence of the presence of most iridoids,including encommiol,genipin,geniposidic acid, geniposide,aucubin,et al.Over the last decade,the antimicrobial,cardiovascular system,immune system,digestive system,sex hormone use and anticancer of Eucommia ulmoides Oliv.have been found and proved by clinical testing.The price of Eucommia ulmoides Oliv.is becoming more and more expensive with the increasing demand from the consumption markets.At present Eucommia ulmoides Oliv.in wild resource mainly.As a result of absoluteness digging,Eucommia ulmoides Oliv.of lots of areas were close to deracinate.In addition,growth cycle of Eucommia ulmoides Oliv.is long,thus comeback of yield cound not act immediately.So it is necessarily to find a new medicine resource.The objective of this study was to initiate a Eucommia ulmoides Oily.hairy root culture using the disarmed agrobacterium tumefaciens strain C58C1 and yield iridoids(aucubin).This study showed that aseptic seedling systems establish,transformed systems establish,the choiceness hairy root lines select,research of hairy root' microstructure and mineral elementBased on the establishment of aseptic seedling systems of Eucommia ulmoides Oliv.,and mediated by the disarmed agrobacterium tumefaciens strain C58C1 which harbors agrobacterium rhizogenes Ri plasmid pRiA4,hairy roots were obtained.The conditions of transformation were optimized.Among different parts explants,such as leaves,cotyledons,and embryonic axis, embryonic axis had the highest induced rate as 76.67%.Immerged by bacteriumand 20 minutes and co-cultured with agrobacterium rhizogenes for 3 days had the highest inducing rate(80%,73%).They were selected in the half-strength MS medium added 500mg/L Cefotaxime.Polymerase chain reaction(PCR)analysis of rolB,rolC concentration analysis were performed.PCR primarily confirmed the integration of rolB,rolC.Among of the yield and contents of acucubin and biomass of twelve hairy root lines,E₁ lines presented the significantly higher content of aucubin(32744.231μg/g).Though research of hairy root's microstructure and mineral element,confirm that the structure characteristic and growth trait are the same as natural root.Meanwhile,there is less starch grain in hairy root confirm why hairy root can grow antrorse,as the statolith theory.Though research of hairy root' mineral element content and natural root',the metabolize power of hairy root is thinner.High level of aluminium element is harmful to succesive transfer culture of hairy root.The establishment of aseptic seedling and plant regeneration systems of Eucommia ulmoides Oliv.,the transformation system based on Eucommia ulmoides Oliv.hairy roots,the selected higheriridoids-production hairy root line give a new pathway to resolve the shortage of iridoids of Eucommia ulmoides Oliv.In addition,the research of hairy root'structure can offer some theoretics for other researchers.