SSR, 2 SSR, 150 bp (Bn92A),250bp (Bn72A). , nitens ; 2.

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1 41 1 () Vol. 41 No Journal of Xiamen University (Natural Science) Jan : (2002) SSR,,, (, ) :, GEN E BAN K, 2 SSR (Bn92A,Bn72A). PCR ( Touchdown PCR),,,, 12 DNA PCR.,PCR SSR,150 bp (Bn92A),250bp (Bn72A). SSR.,,2 SSR. :SSR ; ;; : Q 753 : A SSR ( simple sequence repeats) DNA [ 1, 2 ] DNA,16,( GT) n,( GA TA) n., PCR,,SSR,,, Dietrich [3 ] SSR, 1. 1cM ( Centi2Morgen). Akkaya [4 ] 96,29 SSR, RFL P 23.,,SSR RFL P. SSR SSR. DNA, DNA,, SSR.,,.,SSR, [5,6 ].,,,GEN E BAN K.,.,, SSR, SSR,, SSR : 2 58, 63 ; 23 # ; 2 98 ; 227,5 ; 2, ; 2RS21, YP1 ; 2 Moricandia nitens ; 2. : : ( ) :( ),,.

2 90 () : Taq DNA (dn TPs) Marker Promega ; SSR 2Bn92A (A) 28,Bn72A ( TAA) 5 ( GA) 4 [ 5,6 ] ;. : PCR : PE,9600 ; :752 ;:B IO2RAD,Power300. :UVP, GDS DNA 5 g,, 50 ml, 25 ml DNA (0. 1 mol/ L Tris2HCl, p H 8. 0 ; mol/ L ED TA, p H 8. 0 ; 1 mol/ L NaCl ; mol/ L 2ME) 20 % SDS 2. 5 ml, min,, 5 mol/ L KAc5 ml,, 30 min g 10 min,,,4 000 g 10 min., 10 mmol/ L N H 4 Ac 70 % DNA g 10 min,. 2 ml TE, DNA. 15 L RNaseA (10 mg/ ml ), 37 1 h,,, h DNA. DNA,70 %, 30 min,. 400L TE, 4. DNA TE 260 nm 280 nm, DNA, % DNA, DNA PCR PCR Touchdown PCR, : 1 PCR buffer, mmol/ L MgCl 2, mmol/ L dn TP,1. 0mol/ L SSR,1 U Taq,30 ng DNA. 20 L. : 94 1 min, 30 sec,72 45 sec, 68,1, ,20., (10 mmol/ L NaOH,95 % formamide,0. 05 %,0. 05 % ), 5 min,. 4 %6 %,100 W 0. 5 h,5l, 100 W 1. 5 h.,, 1 L (10 % ), 30 min., 1L ( 1 g/ L AgNO 3, % Formaldehyde). 30 min,, 1 L (2. 5 % Na 2 CO 3, % Na 2 SO 3, % formaldehyde), 1 L,3 min DNA Tab. 1 The results of the DNA extraction of 12 plant samples A 260 A 280 A 260 / A 280 (g/ g FW) # RS YP Moricandia nitens

3 1 :SSR DNA (112 :1. 58 ; ; 3. 3 # ; ; ; 6. 5 ; 7. ; 8. ; 9. RS21 ; 10. YP1 ; 11. Moricandia nitens ; 12. ; M :DNA/ Hind ) Fig. 1 DNA electrophosis examination of 12 plant species 2 SSR 12 (112 :1. 58 ; ; 3. 3 # ; ; ; 6. 5 ; 7. ; 8. ; 9. RS21 ; 10. YP1 ; 11. Moricandia nitens, ; 12. ; M :1kb ) Fig. 2 Amplification of 12 different species DNA with SSR primers of B rassica. napus, PCR DNA 12 DNA A 260 A 280 1, 1. 1, 12 DNA A 260 / A ,DNA. 1, DNA,23 kb. 12 DNA PCR SSR PCR Bn92A Bn72A SSR 12 DNA PCR, 2. 2, Bn92A,(, ) ; (RS21, YP1) ; ( Moricandia nitens), ()2 ; ( 58, 63), (27,5)2; (3 # )2 ; ( 98)2,150 bp. Bn72A YP1,

4 92 () bp.. [13,14 ] SSR Bn92A Bn72A. SSR,., SSR. 3,., 648 [ 7 ]., [8,9 ] [10,11 ] [1215 ] [16 ] [17 ] [18 ],2,.,,,,.. SSR,.. Asya [19 ],() SSR.,,,. : [1 ] Litt M, Luty J A. A hypervariable microsatellite reeveae by in vitro amplification of a dinucleotide repeat within the cardiac muscle actin gene [ J ]. Am J Hum Genet, 1989, 44 : [ 2 ] Love J M, Knight A M. Towards construction of a high2 resolution map of the mouse genome using PCR2analyzed microsatellites [ J ]. Nucleic Acids Res., 1990, 18 : [3 ] Dietrich W F, Miller J, et al. A comprehensive genetic map of the mouse genome[j ]. Nature, 1996, 380 : [4 ] Akkaya M S, Cregan P B. Length polymorphisms of simple2sequence repeat DNA in soybean [ J ]. Genetics, 1992, 132 : [ 5 ] Kresovich S, Szewe2Mcfadden A K. Abundance and char2 acterization of simple2sequence repeats ( SSRs) isolated from a size2fractionate genomic library of B rassica napus L. (rapeseed) [J ]. Theor Appl Genet, 1995, 91 : [ 6 ] Szewc2Mcfadden A K. Kresovich S. Identification of polymorphic conserved simple2sequence repeats ( SSRs) in cultivated Brassica species [ J ]. Theor. Appl. Genet, 1996, 93 : [ 7 ] Obrien S, Seuaneze H N, Womax J. Mammalian genome organization: An evolutionary view [ J ]. Annu. Rev. Genet, 1988, 22 : [8 ] Bonierbale M W, Plaisted R L, Richter s,et al. RFL P maps based on a common set of clones reveal modes of chromosomal evolution in potato and tomato [J ]. Genet2 ics, 1988, 120 : [9 ] Tanksley S D. High density molecular linkage maps of the tomato and potato genomes : biological inferences and practical implications[j ]. Genetics, 1992, 132 : [10 ] Prince J, Pochard E, Tanksley S D. Construction of a molecular linkage map of pepper and a comparison of syteny with tomato [J ]. Genome, 1993, 36 : [11 ] Tanksley S D, Bernatzky R. Conservation of gene repertoire but not gene order in pepper and tomato [J ]. Proc Natl Acad Sci. USA, 1988, 85 : [12 ] Guimares C. Comparative mapping of sugarcane, maize and sorghum. Plant Genome IV [ M ]. San Diego, USA. : Garland Publishing, [13 ] Hulbert S H, Richter T E. Genetic mapping and char2 acterization of sorghum and related corps by means of maize DNA probes[j ]. Proc. Natl. Acad. Sci. USA, 1990, 87 : [14 ] Periera M G, Lee M, Bramel P. Construction of an RFL P map in sorghum and comparative mapping in maize[j ]. Genome, 1993, 37 : [15 ] Witkus R, Womak E. Comparative genome mapping of

5 1 :SSR 93 sorghum and maize[j ]. Genetic, 1992, 132 : [ 16 ] Ahn S, Tanksley S D. Comparative linkage maps of the maize and rice genomes[j ]. Mol. Gen. Genet, 1993, 247 : [17 ] Ahn S, Anderson J A. Homoeologous relationships of rice, wheat and maize chromosome [ J ]. Mol. Gen. Genet., 1993, 241 : [18 ] Teutonico R A, Osborn T C. Mapping of RFL P and qualitative trait loci in B rassica rapa and comparision to the linkage maps of B. napus, B. oleracea and A ra2 bidopsis thaliana[j ]. Theor. Appl. Genet, 1994, 89 : [19 ] Asya R, Yujuni K. Family of crucifurous[j ]. Planta, 1997, 34 : Common SSR Primers for Molecular Marker in Different Crops ZHOU Han2tao, ZHEN G Wen2zhu, ZHOU Yi2ting, XIA Tao ( School of Life Sciences, Xiamen University, Xiamen , China) Abstract : Using 2 pairs of SSR (Simple Sequence Repeats) primers form B rassica napus2bn92a Bn72A, syn2 thesized according to the reports on GEN E BAN K, DNAs extracted from 12 different crops (rice, maize, cot2 ton, rape, orange, cabbage, et al. ) were amplified bytouchdownpcr. The result shows that, the main bands of 150 bp and 250 bp amplified by Bn92A Bn72A respectively appear in all 12 crops and are the same size in B rassica napus by the same SSR primers. The results indicated that different kinds of genomes of crops have the certain extent comparability, and can use the common SSR primers for the molecular markers on genome comparative mapping. Key words : SSR ; molecular marker ; genome comparative mapping ; crops