6,2,1998 5 CHIN ESE BIODIV ERSIT Y 6 (2) :99104, May, 1998 Ξ RAPD 1 1 2 1 1 2 1 (, 100081) 2 (, 430062) 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 100081 2) Institute of Oil Crop, Chinese Academy of Agricultural Sciences, Wuhan 430062 ( B rassica rapa), RFL P ( Restriction Fragment Length Polymorphism) [1 ], B. rapa :, ; [2, ],,,, RFL P RAPD, [3 ],,, ; RFL P,, RAPD DNA [4 ],90, Ξ :1996-07 - 09 ; :1997-08 - 04 Ξ
100 6 [57 ] [8,RAPD ] : [9 ] ;Jain [10 ] [11 ;Demeke ],,, 34,, 1 111 34 (1),, 5, 170,4,, - 70 1 Table 1 Plant materials Order Ξ No. Acc. Cultivars of B rassica rapa 1 0264 (Changde sweet) 2 0265 3 0266 (Lixian Changsha yellow) 4 0267 5 0268 6 0269 7 0270 (Cili white) 8 0271 (Cili foreign) 9 0272 (Dayong flowery) 10 0312 (Quanhu) 11 0313 12 0314 (Changsha Yigu fragrant) 13 0315 (Changsha Guizhou zi) 14 0316 15 0317 16 0318 17 0319 18 0320 19 3675 (Songluo small) 20 1742 21 1743 22 1745 23 1746 24 1747 (Suixian earth) 25 1748 26 1749 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)
2 : RAPD 101 1 () Table 1 (continued) 27 1767 (Zhongxiang native) 28 1769 ( Yichang partition) 29 3640 (Shimen flowery seed) (Anhui) 30 3892 3 (Simao No. 3) ( Yunnan) 31 3893 4 (Simao No. 4) ( Yunnan) 32 1386 (Short big pole) (Zhejiang) 33 1387 (Doujiang) (Zhejiang) 34 3894 (Black leaves Chinese cabbage) ( Guangdong) Ξ Number of accession incatalog of Oilseed Brassica Cultivars in China 112 [12 11211 DNA Colosi Schaal ], 0. 1 g, [13 4 mm, DNA Hillis ],CTAB, DNA 100 ml TE (p H 8. 0), DNA 11212 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 ( ) ;0. 625 TaqDNA ( Promega ) 25 ng DNA ;PTC2100 DNA ( J M Research ) :94 1 min,38 5 min,72 2 min,38 72 3 min ( [9 0. 2 ),45 ] 11213 DNA 1 TAE, 1. 5 %, 100 (bp) DNA (1001500 bp) ( GIBCO2 BRL ) 11214,Ward s 2 211 RAPD DNA RAPD CTAB DNA PCR,, DNA 212 RAPD, 2 DNA, :1) DNA ;2) 2 ;3) DNA 3 40, 19 DNA (2) 213 PCR 19 34 DNA PCR (1) DNA, 23, DNA, RAPD, 200 bp 2000 bp DNA,19, 117 DNA, 102
102 6 2 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 214 1 19 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 1 2 3 4 1 2
2 : RAPD 103 3 2 34 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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