2007, 34 (2) : 305-310 Acta Horticulturae Sinica SSR 1, 1, 1, 2, 1, 1, 23 ( 1, 125100; 2, 100093) : SSR 41, 12 SSR 114, 915 011517 017079, 014630 (BGT23b CHO2D11) 41 2, ; ;,,, : ; ; ; SSR : S 66112 : A : 05132353X (2007) 0220305206 SSR Ana lysis of Genetic D iversity of Pear Cultivars CAO Yu2fen 1, L IU Feng2zhi 1, GAO Yuan 1, J IANG L i2jie 2, WANG Kun 1, MA Zhi2yong 1, and ZHANG Kai2chun 23 ( 1 Research Institute of Pom ology, Chinese A cadem y of A gricultural Sciences, X ingcheng, L iaoning 125100, China; 2 Institute of Forestry and Pom ology, B eijing A cadem y of A griculture and Forestry Sciences, B eijing 100093, China) Abstract: Genetic diversity of 41 pear cultivars was analyzed by using SSR markers. Twelve SSR markers amp lified 114 alleles, w ith an average of 915 alleles per locus. Locus heterozygosity was 011517-017079, and the genetic diversity index was 014630. Two pairs of p rimer (BGT23b and CHO2D11) could distinguish all of the pear cultivars excep t the mutant cultivars. UPGMA cluster analysis showed that the cultivars could be classified into two group s, the occidental pear and oriental pear. Py rus bretschneideri Rehd., P. ussu riensis M axim., and some P. py rifolia ( B urm. f. ) Nakai cultivars originated in China interveined each other; Kuerlexiang and other P. sinkiangensis Y varieties could be clustered together. Chi2 nese P. py rifolia Baozhu and Deshengxiang, Japanese P. py rifolia Shinseiki and Housui, Korean P. py rifolia W hangkeum bae were clustered together in the dendrogram. The coefficient between Q iubai and M ili was high, and they belonged to the same population, so as Nanguo, M anyuanxiang, Q iuzi and B alixiang. J inhua origina2 ted correlatively w ith J inchuanxue and Cangxixue. Key words: Pear; Cultivar; Genetic diversity; SSR ( Pyrus L. ) (Bailey, : 2006-07 - 11; : 2006-12 - 04 3 Author for correspondence ( E2mail: zhangkaichun@baafs1net1cn) 1917), ( P. bretschneideri Rehd. ) [ P. pyrifolia (Burm. f. ) Nakai] ( P. ussuriensismaxim. ) ( P. comm unis L. ) ( P. sinkiangensis Y ), ( Yamamoto et al., 2001, 2002a, 2002b) SSR SSR,
306 34 1 41, ( 1) 1 Table 1 Cultivars of pear for SSR ana lysis ( ) Species (parentage) Code Cultivar O rigin ( ) Species (parentage) Code Cultivar O rigin 1 22 P. bretschneideri Dangshan Suli Dangshan, Anhui Dananguo L iaoning Rehd. 2 23 Jinchuanxue Jinchuan, Sichuan P. pyrifolia Baozhu Chenggong, Yunnan 3 (Burm. f. ) 24 Jinli W anrong, Shanxi Nakai Deshengxiang Sichuan 4 25 Lanzhou Dongguo Longzhong, Gansu Cangxixue Cangxi, Sichuan 5 26 M ili Hebei Shinseiki Japan 6 27 Pingguoli Yanbian, Jilin Housui Japan 7 28 Q iubai Hebei Whangkeumbae Korea 8 29 Xuehua D ingxian, Hebei P. sinkiangensis Kuerlexiang Kuerle, Xinjiang 9 Y 30 Chili Chip ing, Shandong Kunqieke Yecheng, Xinjiang 10 31 Jinfeng Xingcheng, L iaoning Hongnahe Tulufan, Xinjiang 11 32 Jinhua Jinchuan, Sichuan Zaoshujuju Kuche, Xinjiang 12 33 Yali Hebei P. comm unis L. Bartlett B ritain 13 34 Chuizhi Yali Shijiazhuang, Hebei Flem ish BeautyBelgium 14 35 Jinxian Dayali Jinxian, Hebei Clapp Favorite America 15 36 P. ussuriensis Jianba L iaoning Yali Bartlett W ujiuxiang Xingcheng, L iaoning Maxim. 16 37 Huagai L iaoning Nanguo Bartlett Jinxiang Xingcheng, L iaoning 17 38 Manyuanxiang L iaoning Kuerle xiang Xuehua Yuluxiang Taiguo, Shanxi 18 39 Q iuzi Beizhen, L iaoning Pingguoli M ishirazu Zaosu Xingcheng, Liaoning 19 40 Balixiang L iaoning Clappπs Favorite Zaosu Bayuehong Meixian, Shaanxi 20 41 Daxiangshui L iaoning Xuehua Shinseiki Huangguan Shijiazhuang, Hebei 21 Nanguo Anshan, Liaoning
2 : SSR 307 CTAB DNA, 1, DNA SSR ( Gianfranceschi et al., 1998; Yamamoto et al., 2001, 2002a, 2002b) Sangon 22 12 SSR : 20 L 15 20 ng DNA, 10 buffer 210 L, MgCl 2 115 mmol/l, dntp 012 mmol/l, 015 mol/l, Taq 1 U PCR PE PTC100 PCR 94 4 m in; 94 1 m in, 61 58 1 m in, 72 2 m in, 10, 015 ; 94 1 m in, 1 m in, 72 2 m in, 30 ; 72 10 m in; 4 6%, SSR 1, 1, 1, 0 h = 1 - x 2 i, x i i, H = h / n, n NTSYS2pc211, D ice (UPGMA ) 2 211 SSR 12 SSR 41, 114, 3 15, 915 ( 1) F ig. 1 BGT23b 41 1 M: pbr322 DNA /M SP1 markers 1 Am plif ica tion of pr im er BGT23b in 41 pear cultivars The same code as in Table 1. M: 011517 017079, 014630 ( 2) 212 1 DNA,,, SSR, BGT23b 41 15, 30 SSR, BGT23b CHO2D11, 41 12, pbr322 DNA /M SP1 markers. 2 12 SSR 41 Table 2 Amplif ied results of 41 pear cultivars with 12 SSR primers Primer Annealing ( ) Number of alleles detected h Locus heterozygosity KU10 60 55 6 013403 KA4b 60 55 4 011553 KA14 60 55 5 012540 KA16 60 55 14 017079 KB16 60 55 3 011517 BGT23b 60 55 15 015848 NH004a 60 55 12 015140 NH015a 61 56 14 015509 NH009b 58 53 8 015235 NH013a 60 55 8 016401 NH017a 61 56 10 014961 CHO2D11 58 53 15 016371 Average 915 014630
308 34 015882,, ;, 015000 015714 015714,,,,, 015405 015556, 213 12, D ice UPGMA ( 2) 41 0 1100, 0133 0128,, ;,, ( ) ( ), ( ),, 3 F ig. 2 1 2 D endrogram of pear cultivars ba sed on SSR da ta The same code as in Table 1. 12 SSR ( ) ( ), Yamamoto (2001) ( O sa Gold
2 : SSR 309 ) ( ), SSR, SSR (Clapp Favorite) ( ) 12 SSR, D ice,,,,, (1946), ; Teng Tanabe ( 2004), P. pyrifolia var. sinensis (L indley) Y. Teng et K. Tanabe,, 016857, 015455, 015294 015455 015294 015806, 015405 015882;,, 015405 015556,,, (, 1963; Teng et al., 2001), 3, 015000 015714 015946,,, Teng (2001) 3, (, 2001), 415% (, 1983;, 1985;, 1986) NH015a NH015a CHO2D11 3, 12 1 2,,, SSR 20,,, SSR,,, References Bailey L H. 1917. Standard cyclopedia of horticulture. New York: Macm illan: 2865-2878. Chen Rui2yang, L i Xiu2lan, TongDe2yao, L iu Yu2xin, W u Q iu2qin, W ang Feng2shan, Pu Fu2shen, Huang L i2sen. 1983. Study on the chromo2 some number of pear ( Pyrus L. ) in China. Acta Horticulturae Sinica, 10 (1) : 13-15. ( in Chinese) 1,,,,,,,. 1983.., 10 (1) : 13-15. Gianfranceschi L, Seglias N, Tarchini R, Kom janc M, Gessler C. 1998. Simp le sequence repeats for the genetic analysis of app le. Theor. Appl. Genet., 96: 1069-1076. Huang L i2sen, L i Shu2ling, Pu Fu2shen. 1986. Study on the chromosome number of pear cultivars in China. China Fruit, (1) : 12-13. ( in Chinese)
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