% 8 F ! 8 y # F _ 栗孟飞,李null 唯*( j ! = (= c ), 50 d s Y 33null 17a12null97a4null 55nullg/mLb null y % 8 F ! 8 , % 8 ! 3 F V b1 o M :% 8 ; ? ; F ; HPLC m s | :R286null 1null null null D S M :A null null null c I | : 0253null2670( 2010)08null1366null05Establishment of tissue culture system of Sinopodophyllum emodiand identification of podophyllotoxinLI Mengnullfei, LI Wei( Collegeof Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, China)Abstract: Objectivenull To establish the tissueculture system of Sinopodophyllumemodi andtheextracnulltion and identification methods of podophyllotoxin. Methods null The effects of seedling induced rate bydifferent concentrations of hormones and activated charcoal ( AC) were studied, taking the separated manullture embryo as material; Then the effect of growth and development of excised rhizome (2. 0 cm) of theseedling subnullcultured were conducted by different concentrations of hormones; In addition, the contents ofpodophyllotoxinin rhizome, culturedmedia, and leaf (leafstalk) were extractedandidentified by HPLC atregular stages. Resultsnull After 20 dthe seedling induced rate reached92.0% with better growth anddevenulllopment with the separatedmature embryo of S. emodi seeds cultured on the Murashige andSkoogemedinullum (MS) supplemented with 2.0% sugar, 0.4% agar, 1. 0 mg/ mL GA3, and 1.0 g/L activated charcoal(AC); After 40 d subnullculture of excised rhizome ( 2. 0 cm), the subnullcultured plantlets showed bettergrowth and development with root length 11. 59 cm and the cultured medium was MS supplemented with2.0% sugar, 0.4% agar, 1.0 mg/ mL IBA, and1. 0g/ L AC, In addition, thepodophyllotoxin indifferentparts of S. emodi including cultured media at different growth stages was extracted and identified byHPLC. The results indicated that the content of podophyllotoxin was rhizome cultured media leaf(leafstalk) with 33.17, 12.97, and 4.55 nullg/ mL, respectively after 50 d. Conclusionnull The tissue culturesystem of S. emodi could be established andthe method of production podophyllotoxin by root culture ofS. emodi is feasible.Key words: Sinopodophyllum emodi (Wall. ) Ying; root subnullculture; podophyllotoxin; HPLCnull null % 8 Sinopodophyllum emodi ( Wall.)Ying l S % 8 M 3 , 1 s S 1 , S W 0 , 1 e b a A e a 5 2 , M ? C ,%8 # 1 c ! a a0 a , ! F ( podophyllonulltoxin) K ( 2null7 4null2%)3b1 King Sullinullvan F l 6 aT , R 1 ,i B D a3 4b F ! = (= c ); H 10 50 d= , a= (= c ) # ! F ( v 9 F , ! 50 d ,F G Q 33null17a12null97a4null55 nullg/ mLb V , K Q 3 F ;6 , 5 ! F A , V 3 ? V 9 | F d b ! , V % 8 $ , % 8 ? ! F 4 | b4null ) null null % 8 F % ! 1 Z ,S= X , Y V F 10a% ! Z E 4 | 3 F 11,13 ,r % % 8 7 3 C bVnull1369null中草药null ChineseTraditional andHerbal Drugsnull 41 8 2010M 8V 1null 3 H % 8 a= (= c ) # ! F Table 1null Content of podophyllotoxininroot, leaf (leafstalk),and culturedmedium of S. emodi at different stages null F / ( nullg nullmL- 1)10 d 20d 30d 40d 50d 4null22# 0null 25 a 7null 52# 0null 07 a 12null36 # 0null23a 22null90 # 0null14a 33null17# 0null 85 a= (= c ) 1null02# 0null 14 b 1null 63# 0null 23 b 3null23 # 0null32c 4null02 # 0null22c 4null55# 0null 62 c ! 1null31# 0null 11 b 3null 22# 0null 49 b 7null49 # 0null22b 9null81 # 0null23b 12null97# 0null 38 bnull null 3 V U Duncan(s k A ( P Z + , null , S i , null % 8 Z Jnull 0 , 2004, 35(1): 98null100null2 null null , null , a; o null 2 % 8 L . DALDs Jnull 0 , 2009, 40(6): 951null955null3 null , S * , + , null HPLC E F 3 0 F ! c Jnull S 0 S v , 1996, 27(4):219null222null4 null King L S, Sullivan Mnull Similari