Establishment Of Transform System In Scutellaria Viscidula Bunge And Its Flavonoids Production

Scutellaria viscidula Bunge is a very old and well-known drug in traditional Chinese medicine, which belongs to the genus of Scutellaria, Labiatae. The high physiological activity of the root extracts is a consequence of the presence of most flavonoids, including baicalin,wogonin,wogonoside,et al.Over the last decade,the antimicrobial,hypotensive,sedative,antioxidant,antitumor properties of baicalein have been found and proved by clinical testing. At the same time baicalein has an inhibitory effect on human immunodeficiency virus(HIV) and HBV.The price of baicalin is becoming more and more expensive with the increasing demondment from the consumption markets.At present baicalin roots in wild resource mainly.As a result of absoluteness digging, Scutellaria viscidula Bunge of lots of areas were close to deracinate. Shanxi, Shandong,Henan,Sichuan have planted artificially, but the output was less.In addition, growth cycle of Scutellaria viscidula Bunge 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 Scutellaria viscidula Bunge hairy root culture using the disarmed Agrobacterium tumefaciens strain C58C1 and yield flavonoids. This study showed that In vitro systems establish, transformed systems establish, the choiceness hairy root lines select and the optimal culture condition of hairy roots systems establish.Based on the establishment of In vitro systems of Scutellaria viscidula Bunge, 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, stems and callus, stems had the highest induced rate as 88.9ï¼…. Immerged by bacteriumand 15 minutes and co-cultured with Agrobacterium rhizogenes for 2 days had the highest inducing rate(84ï¼…, 82ï¼…).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 flavonoids and baicalin of eight hairy root lines, Hq4 lines presented the significantly higher content of flavonoids (44.58 mg/flask) and higher content of baicalin(22.49 mg/flask).The effects of different mediums and exogenous phytohormones on hairy roots growth and biosynthesis of flavonids in the hairy root cultures of Scutellaria viscidula Bunge were studied. The results showed that contained 30g/L sucrose, 1/2MS with pH 5.8 was best medium and the 2,4-D,NAA,6-BA and GA all have obvious effects on the growth of hairy root cultures and the biosynthesis of flavonids.The growth of hairy roots and biosynthesis of flavonids were strongly restrained by 2,4-D. However, NAA,GA and 6-BA of appropriate concentration were favourable to hairy roots growth and flavonids production and the effect of GA was prominence.The establishment of In vitro and plant regeneration systems of Scutellaria viscidula Bunge, the transformation system based on Scutellaria viscidula Bunge hairy root, the selected higher-baicalin-production hairy root line and eminent Cultured-conditions of hairy root give a new pathway to resolve the shortage of baicalin of Scutellaria viscidula Bunge.