Studies On Hairy Roots Induced Scutellaria Baicalensis Georgi And Separation Purification Of Baicalin

Dried root of Scutellaria baicalensis Georgi，a member of Labiatae，is considered as oneof well known drug in Chinese medicine(TCM)witll many pharmacological effects.Scutellaria baicalensis Georg’s nature is cold,gout is bitter. Scutellaria baicalensis Georgwere repored that Baicalin had multiple biological activities such as effect heat anddampness,detoxification,bleeding tocolysis.There are many Scutellaria baicalensis Georgisgrow in the sunny slopes,forest edge and sparse grass.They are heat,cold-resistant,droughtand avoid water. They can through winter at-35℃a nd resistance high temperature at35℃.The main active ingredient of Scutellaria baicalin (Baicalin), is extracted fromScutellaria root out of a separation of flavonoids, has significant biological activity. In recentyears, Medicinal Scutellaria increased year by year, limited resources have been a wildpredatory excavation, shortage of resources to high prices. The use of hairy root cultures ofScutellaria baicalin production will achieve industrialization, has an important economicvalue.Using genetic transformation and plant tissue culture technology, Hairy Root ofScutellaria baicalensis Georgi was induced by Ri-Plasmid of Agrobacterium rhizogenesstrains1.2556and co-cultured. Effects of different factors such as explants、preculture time、infecting time、Acetosyringone on frequency of transformation of Scutellaria baicalensisGeorgi were studied.Based on the use of Box-Benhnkcn design and response surface analysisof the incubation time, temperature, rotation speed, inoculum culture conditions and otherfactors that affect the analysis in the single-factor experiment then skullcap hairy root culturesystem in vitro. Baicalin of Hairy Root of Scutellaria baicalensis Georgi was induced byRi-Plasmid application of macroporous resin for separation and purification then detected byHPLC.Studies have shown that achieved by co-culture method skullcap Genetic transformationof hairy roots. The hairy root growed densely and horizontally and was ageotropism. Hairyroot was achieved in MS medium without any plant horomons.Explants skullcap stems, rootsare able to produce hairy roots, but induced the highest rate of stems, up to47%. The optimalconditions for induction: two days pre-culture of Explants. infection time of10-15min add100μmol/L AS, Co-culture two days. The transformation frequency could be raised by culturemedia with400mg/L Ceftriaxone. The optimum liquid medium for hairy root of Scutellaria baicalensis Georgi wasobtained.1/2MS medium was determined by the comparison of hairy root fresh weight. Theresult showed that the optimum condition were1/2MS medium, sucrose30g/L, pH6.0，incubation time25d,temperature27.4℃, rotation speed123r/min, inoculum0.5g as rotationalspeed. Conducive to the growth of hairy roots and the biosynthesis of baicalin in the dark.By macroporous resin on hairy root extract separation and purification, to determine thehairy root extract contains baicalin. Purified using HPLC for detect baicalin showed thatbaicalin of hairy roots content of up to5.37%in growth of25days.