Studies On Induction Of Arnebia Euchroma (Royle) Johnst Hairy Roots

Arnebiaeuchroma(Royle)Johnst is a plant with wide uses. Shikon in derivatives,extracted from Arnebiaeuchroma(Royle)Johnst, were extensively applied in medical treatment,food and other f ields.Agrobacterium rhizogenes were employed to induce hairy roots from explants of Arnebia euchroma (Royle) Johnst . Effect of Agrobacterium rhizogenes,OD₆₀₀,days shoot,explant,medium and AS on hairy root induction was investigated.1. The effects on the cotyledons of Arnebiaeuchroma(Royle)Johnst with different concentration of kanamycin and hygroycin were investigated. The results indicated that the quality concentration of kanamycin for selecting the cotyledons was 60mg/L and adequat concentration of Cefotaxime was 600-1000mg/L. The explants were subcultured every 12d.If the concentration of Cefotaxime changed higher, the subcultured time would be shortened.2.The hairy roots were induced from infected cotyledon explants of Arnebiaeuchrom(Ryl- e)Johnst when the concentration of Agrobacterium rhizogenes MSU440 was OD₆₀₀0.5.The red hairy roots could grow rapidly and stabilitly on hormone free medium.The result of PCR showed that rolc gene in Riplasmid was integrated into genome of Arnebiaeuchroma(R- oyle)Johnst.3. The hairy roots in B5 medium were subcultured every 25-30d. The growth rate of hairy roots was 7～9 times in one month. A technical system of preservation hairy roots in solid medium were established.4. Hairy roots were obtained by using Agrobacterium rhizogenes MSU440 to infect five explants of the Arnebiaeuchroma(Royle)Johnst for the frist time.There are some significant characteristics of the hairy root such as growth in hormone free media,fast growth capacity,more branches,and more root hairs. A culture system for hairy root was preliminarily established. It is the basis for producing shikonin by using hairy roots.