RAPD. ( B rassica rapa) RFL P ( Restriction Fragment Length Polymorphism) RFL P ; RFL P, rapa. CHIN ESE BIODIV ERSIT Y 6 (2) :99 104, May, 1998 RAPD

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1 6,2, CHIN ESE BIODIV ERSIT Y 6 (2) :99104, May, 1998 Ξ RAPD (, ) 2 (, ) RAPD, 34,Ward s : ; ;DNA 34 8,, RAPD Genetic diversity among Chinese oilseed accessions of Brassica rapa as determined by RAPD markers/ ZHU Li 1), L I Ru2Gang 1), WU Xiao2Ming 2), WU Ning2Feng 1), FAN Yun2Liu 1), QIAN Xiu2Zhen 2) Abstract The genetic diversity among 34 Chinese oilseed accessions ( mainly from Hunan and Hubei Provinces) of B. rapa was analyzed by RAPD technique and statistical methods. A general molecular method for assessment of genetic diversity was established from this research. Cluster analysis showed that the genetic variation among oilseed accessions of B. rapa was closely related with their eco2geographic distribution ; exten2 sive variation existed among the accessions from Hunan and Hubei Provinces ; 34 accessions were divided into eight groups at DNA level. The application of results to selection of parents in breeding program was dis2 cussed. Key words genetic diversity, oilseed B rassica rapa, RAPD Author s address 1) Biotechnology Research Center, Chinese Academy of Agricultural Sciences, Beijing ) Institute of Oil Crop, Chinese Academy of Agricultural Sciences, Wuhan ( B rassica rapa), RFL P ( Restriction Fragment Length Polymorphism) [1 ], B. rapa :, ; [2, ],,,, RFL P RAPD, [3 ],,, ; RFL P,, RAPD DNA [4 ],90, Ξ : ; : Ξ

2 100 6 [57 ] [8,RAPD ] : [9 ] ;Jain [10 ] [11 ;Demeke ],,, 34,, (1),, 5, 170,4,, Table 1 Plant materials Order Ξ No. Acc. Cultivars of B rassica rapa (Changde sweet) (Lixian Changsha yellow) (Cili white) (Cili foreign) (Dayong flowery) (Quanhu) (Changsha Yigu fragrant) (Changsha Guizhou zi) (Songluo small) (Suixian earth) Origin ( Yuanjiang Guilin zi) ( Hanshou white) ( Hanshou Nanjing zi) ( Hanshou Wanshou County sweet) ( Youxian premature short) (1) ( Yueyang sweet 1) (2) ( Yueyang sweet 2) ( Yueyang black zi) ( Hengyang) ( Hengyang sweet) ( Yicheng da) ( Yicheng white) (1) (Suixian da 1) (2) (Suixian da 2) ( Echeng white) ( Echeng sweet)

3 2 : RAPD () Table 1 (continued) (Zhongxiang native) ( Yichang partition) (Shimen flowery seed) (Anhui) (Simao No. 3) ( Yunnan) (Simao No. 4) ( Yunnan) (Short big pole) (Zhejiang) (Doujiang) (Zhejiang) (Black leaves Chinese cabbage) ( Guangdong) Ξ Number of accession incatalog of Oilseed Brassica Cultivars in China 112 [ DNA Colosi Schaal ], 0. 1 g, [13 4 mm, DNA Hillis ],CTAB, DNA 100 ml TE (p H 8. 0), DNA RAPD 5 DNA, 5 ng/ ml, PCR 25 ml, 10 mmol/ L Tris2HCL (p H8. 0) ;2. 5 mmol/ L MgCl 2 ; 0. 2 mmol/ L da TP,0. 2 mmol/ L dctp,0. 2 mmol/ L d GTP,0. 2 mmol/ L dttp ( BM ) ;0. 64 mmol/ L ( ) ; TaqDNA ( Promega ) 25 ng DNA ;PTC2100 DNA ( J M Research ) :94 1 min,38 5 min,72 2 min, min ( [ ),45 ] DNA 1 TAE, 1. 5 %, 100 (bp) DNA ( bp) ( GIBCO2 BRL ) 11214,Ward s RAPD DNA RAPD CTAB DNA PCR,, DNA 212 RAPD, 2 DNA, :1) DNA ;2) 2 ;3) DNA 3 40, 19 DNA (2) 213 PCR DNA PCR (1) DNA, 23, DNA, RAPD, 200 bp 2000 bp DNA,19, 117 DNA, 102

4 Table 2 Oligonucleotide primers used in the test Primer (5 3 ) Sequence (5 3 ) Primer (5 3 ) Sequence (5 3 ) Primer (5 3 ) Sequence(5 3 ) 1 GCG GCT GGA G 8 A GG ACG TGC C 15 TGC GCC GGT G 2 CCA A GA TGC T 9 CTC CTC CCC C 16 GCG GA G GTC C 3 CAA GGG A GG T 10 GCT CCC CCA C 17 GCG TCG A GG G 4 AA G GCA CCA T 11 TA G GAC A GT C 18 A GT GAC CGG C 5 GTG CGT GGC T 12 CCC GCC GTT G 19 A GC ACG GGC A 6 TGC TA G CCT C 13 CTC TCG TGC G 7 GCA A GT CAC T 14 GCC ACC TCC T DNA Fig. 1 DNA fragments amplified by primer 19 Ξ Ξ Stands for the order of the accessions RAPD, 34 (2) 34 8 (3) 3 8 Table 3 Eight groups of oilseed B rassica rapa Group Cultivar ( B rassica rapa) Origin

5 2 : RAPD Fig. 2 The relationship among 34 accessions of B rassica rapa as determined by Ward s cluster method from RAPD data,,,,,,, 1, ( ) [14 ] ( ) [15 ], ( ) [16 ],,, DNA RAPD 5 DNA

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