The Optimization For The Induction And Growth Of Atractylodis Japonicae Koidz. Ex Kitam. And Celosia Argentea L. Hairy Root And Analysis Of Its Secondary Metabolites

In this paper，the use of Agrobacterium rhizogenes R1000and R1601strains to infectAtractylodis Japonicae Koidz. ex Kitam. explants，the use of Agrobacterium rhizogenesR1,A4,1025,ATCC15834,R1000and R1601strains to infect explants, optimizaed for theinduction and growth of Atractylodis Japonicae Koidz. ex Kitam. and Celosia Argentea L. hairyRoot, established Atractylodis Japonicae Koidz. ex Kitam. and Celosia Argentea L.culturesystem of hairy roots. Then determined the content of polysaccharide and flavone fromAtractylodis Japonicae Koidz. ex Kitam.and Celosia Argentea L.ofsecondary metabolites inhairy root，and its antibacterial role of basic research.The results show that，Atractylodis Japonicae Koidz. ex Kitam. leaves and root wouldproduce hairy roots. While then R1000strains the highest infection ratio, the best inducingmaterial is Atractylodis Japonicae Koidz. ex Kitam. leaves, and the best inductivity is62%±2.58%,0.1%corrosive sublimate disinfect6min of Atractylodis Japonicae Koidz. exKitam.seed. Atractylodis Japonicae Koidz. ex Kitam. explants leaf area cut out1.5×1.5cm2,Rhizoma Atractylodis Japonicae explants pre-culture36~48h, Agrobacterium rhizogenes R1000and R1601infect6min of Atractylodis Japonicae Koidz. ex Kitam. explants. Culture ofAtractylodis Japonicae Koidz. ex Kitam. hairy roots is white, Atractylodis Japonicae Koidz. exKitam. explants age to21d. Celosia Argentea L.’s leaves, stems, cotyledon, regeneration plantand flowers would produce hairy roots by R1, A4,1025, ATCC15834, R1000and R1601.0.1%corrosive sublimate disinfect2min of Celosia Argentea L. seed. MS solid medium pH value of6.0, Celosia Argentea L.leaf area cut out2.0×2.0cm2, Celosia Argentea L. explants pre-culture12~24h, Agrobacterium rhizogenes R1, A4,1025, ATCC15834, R1000and R1601infect4minof Celosia Argentea L. explants, While then A4strains the highest infection ratio, the bestinducing material is Celosia Argentea L. leaves, and the best inductivity is98±1.61%, Culture ofCelosia Argentea L. hairy roots is white, sucrose concentration is4%. PCR results show: rol Bgene of R1000and R1601are integrated into the hairy roots of Rhizoma Atractylodis Japonicaeexplants of leaves and root; rol B gene of A4and R1are integrated into the hairy roots ofCelosia Argentea L. explants of leaves.Atractylodis Japonicae Koidz. ex Kitam. hairy roots polysaccharide content of126.73mg/g,Atractylodis Japonicae Koidz. ex Kitam. hairy roots flavonoids content of4.76mg/g, CelosiaArgentea L. hairy roots polysaccharide content of68.81mg/g, Celosia Argentea L. seeds ofpolysaccharide content of56.64mg/g, Celosia Argentea L. hairy roots polysaccharide is Celosia Argentea L. seeds polysaccharide1.2times, Celosia Argentea L. hairy roots flavonoids content of103.42mg/g, Celosia Argentea L. seeds flavonoids content of60.15mg/g, Celosia Argentea L.hairy roots flavonoids is Celosia Argentea L. seeds flavonoids1.7times.When extracting solution of Celosia Argentea L. hairy roots flavonoids is50mg/ml,Escherichia coli inhibition is47.24%, Salmonella inhibition is46.44%, Pseudomonasaeruginosa inhibition is37.47%, MRSA inhibition is37.24%, Streptococcus inhibition is35.20%. Extracting solution of Celosia Argentea L. hairy roots flavonoids inhibition concentratethe same kind of bacteria:50mg/ml>25mg/ml>12.5mg/ml.