DNA, ( ) , ; 21 , ) H1 yungningensis Hand2Mazz1 : H1 hemsleyanum Diels : H1 cendicans Wall1 ex DC1 :

Transcription

1 H1 yungningensis Hand2Mazz1 : H1 cendicans Wall1 ex DC1 :, H1 dissectifolium K1 T1Fu1 : 1, 2,,, 2 2, H1 hemsleyanum Diels : 3 12, , 5,, , 1,, 2,415 cm,39 cm,, H1 millofolium Diels : 2,3715 cm,1215 cm,,, 3, 113, : Aralia cordata Thunb1 :,,, 1 1 ( ) 1 : 2, Aralia kansaensis Hoo : (,, ) ( ) 1 :, ( ) DNA (11, ; 21, ) : : DNA (RAPD) DNA : DNA, DNA :RAPD, RAPD

2 , (21226,7421,5804,5634,4878,3530 bp) (A2 ( Magnolia officinalis Rehd1 et Wils1) ( M1 garose) Gene TaqDNA dntps officinalis Rehd. et Wils. var. biloba Rehd1 et Wils1),, ;DNA/ EcoRI + Hind DNA Ladder ( 21227, 5148, 4973, 4268, 3530, 2027,1904,1584,1375,947,831,564,125 bp), (SABC) ; (magnolol) (honokiol) 2 ( 2eudesmol) Gene Amp PCR System 2400 ( Perkin, Elmer Corporation) ; Ultraspec 2000 UV/ Visible Spec2, trophotometer ( Pharmacia Biotech) ; TGL216G, ( ) ;DK28D, 6 30 ( ) ; PAC 3000 (Bio2 2, rad Corporation) ; ( ) DNA 1 g,, ml Eppendorf 47 DNA, 1 ml - 20,, DNA 67 s,8000 r/ min 10 s,, DNA(RAPD) %CTAB 1 ml, ,60 4 h, 015 h 14, l : (24 1) 10 min, r/ min 10 min,, DNA(RAPD) DNA 2/ 3 4,,-, %4, , : 11 ( M1 officinalis Rehd1 et Wils1), 21 ( M1 officinalis Rehd. et Wils. var. biloba Rehd. et Wils. Cheng),31 M1 de2 nudata Desr,41 M1 Biondii Pamp,51 M1 sprengeri Pamp,61 M1 liliflora Desr,71 M1 delavayi Franch,81 M1 chingii Dandy,91 M1 insignis (Wall) Bl 101 ; r/ min 5 min,,, 800 l min,3000 r/ min 10 min,, 015 ml Eppendorf, 1 h50 l TE(pH 810) Glass2milk Kit DNA50 l DNA 3, 10 l,, 5 min 8000 r/ min, 125 l, Liriodenron chinense Hemsl Snrg,, RAPD TE (ph 810) 20 l,,60 5, min, r/ min,, (South Chi2 DNA 40 l na Botanical Garden, Guangzhou), 212 DNA, DNA :7 l DNA 3 l 112 DNA, 114 % ( (CTAB) Tris ( ) 110g/ ml),5 V/ cm 1 h,254 nm 2mercaptoethenol ( 2 ) SIGMA DNA DNA/ Hind DNA/ Hind Promega DNA : Pharmacia

3 Biotech Ultraspec 2000,5 l DNA 213 DNA Gene Amp PCR System l, 20, (Perkin Elmer Corporation),, 260 nm 280 nm 320 nm 25 l, 115M, DNA 25 ng/ l,, (A A 320 ) / (A A 320 ) DNA,A A 320 [ (A A 320 ) mm KCl, 10 mm Tris1 HCl, 011 % Triton X2100, ph 50g/ ml ],DNA l/ 1 813) 215 l,110 Unit TaqDNA polymerase ( Sangon Com2 1000, ng/ mg (DNA 50 l TE, 20, 1 g) :95 3 min 20 sec, DNA 10 min A 260 A 280 A 320 ng/ l ng/ mg dntps 200 M,MgCl mm,10 buffer (10 pany) DNA :94 1 min,36 1 min,72 2min, PCR RAPD 115 %, :25 l 5 l, DNA/ EcoRI + Hind DNA Lad2 der 5 l,5 V/ cm 60 min,254 nm 215 ( F) F = 2N XY / (N X + N Y ), N X N Y X Y DNA,N XY X Y DNA 12F, UPGMA, 2 RAPD 1 vs 2 1 vs 3 1 vs 4 1 vs 5 1 vs 6 1 vs 7 1 vs 8 1 vs 9 1 vs 10 S S S S S S S S S S S S S S S S S S S S :11 1 ;

4 3 2 12F ( ) RAPD ( ) / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / : 1 RAPD, 12F UP2 GMA Sangon, 3 MgCl mm,10 buffer (10 mm KCl,10 mm 311 DNA DNA Tris1HCl,011 %Triton X2100,pH 813) 215 l,110 Unit bp Taq DNA DNA,1 DNA DNA, RAPD 118, 40 ng/ mg DNA 10 DNA RNA RNA, 312 DNA 46 TE K DNA, DNA RNA RAPD, DNA DNA DNA,,, DNA 1429,,DNA, DNA 1169, %,DNA RAPD RAPD Williams 1990, DNA, RAPD, Taq DNA PCR Mg 2 + DNA PCR,,RAPD, RAPD,, Taq DNA PCR Mg ng 55 ng 10 ng, RAPD, PCR PCR, Gene Amp DNA, PCR System 2400 ( Perkin Elmer Corporation) DNA 15, Biometera UNO PCR,, ;

5 F 0160, PCR 3 / s, 2 /, s, RAPD, DNA 412 RAPD, 16, ( M1 delavayi) 8 DNA 011 %, DNA 1 3, ( ), ( M1 2, RAPD chingii) ( M1 insignis) DNA, ( M1 of2 17 RAPD, 12F ficinalis) ( M1 officinalis Rehd. et Wils , 1 var. biloba Rehd. et Wils. Cheng),,0185 ( M1 denudata ) ( M1, biondii) ( M1 sprengeri ) ( M1 lil2, iflora) ( M1 delavayi), , 19, ; 0160, RAPD DNA, ( Magnolia) (Subgen Magnolia),,,, 3 : : ( Magnolia ) ( Subgen Pleurochasma), :, ( M1 chingii) ( M1 insignis) 1,1964, ( Manglietia), 3, , 1,1984,19(3) 213 (Liriodenron chinense) 4,1,,RAPD 1,1996,27(1) , 5,1 1, 1992,15(2) ,1 8 ( Iiriodenron chi2 1,1994,19(8) 461 nense), 7,1 1,1992,17(7) 404, ( M1 sprengeri) 8,1 RAPD 7, ( M1 denudata ) ( M1 1,1996, biondii) ( M1 liliflora) 9,1 RAPD 1, ( M1 insignis),,1997,22 72 ( M1 chingii) 10,1RAPD 1, ( M1 delavayi),1998,29(1) ,1 3 RAPD 3 1 ( M1 delavayi) 12 1,1999,24(6)

6 12,1 RAPD 1,1999,24(8) ,1 DNA 1, 1996,19(12) Cao H, et al1 A molecular approach to identification of the Chinese drugpu2gong2ying ( Herba Taraxaci) and six adul2 terant by DNA fingerprinting using random2primed polymerase chain reaction1 J Chin Pharm Sci, 1996, William J G K1 DNA polymorphisms amplified by arbitrary primers are useful as genetic markers1 Nucl Acids Res, 1990,18(22) ,1 RAPD 1 ( ),1999,38(1) ,1 4 1,1994,19(10) ,1 1,1999,22 (8) ,1 1, 1998,21(4) 166 ( ) Studies on Random Amplified Polymorphic D NA Fingerprinting of Cortex Magnoliae Officinalis Liu Wensheng 1, Zhu Jianming 1, He Bin 1, Su Yingjuan 2 (11Guangdong Provincial People s Hospital, Guangzhou ; 21School of Life Sciences, Zhongshan University, Guangzhou ) Abstract Objective : To identify Chinese traditional medicine(ctm)hou2pu(cortex Magnoliae Officinalis), its counterfeits and sub2 stitues1 Methods : Total genomic DNA samples of ten plant species were amplified by randomly amplified polymorphic DNA(RAPD) 1 Results : Ten samples were able to be distinguished through their amplified DNA banding patterns on the agarose gels after electrophoresis1 Conclusion : RAPD is able to identifyhou2pu, its counterfeits and substitutes quickly and truly, which is also quite valuable for correctly introducing plant1 Key words Chinese traditional medicine ; Cortex Magnoliae Officinalis ; RAPD (, ) Ocimum basilicum var1 pilosum (Willd1) Benth1,,, Ocimum basilicum var1 pilosum (Willd1) Benth1,,,,,, ;,,5,,,, ( 2 mm) 1,,,, ; 1 26,, Ocimum basilicum var1 pilosum (Willd1) Benth1, ,15,,