SSR, SSR Ana lysis of Genetic D iversity of Pear Cultivars

Transcript

1 2007, 34 (2) : Acta Horticulturae Sinica SSR 1, 1, 1, 2, 1, 1, 23 ( 1, ; 2, ) : SSR 41, 12 SSR 114, , (BGT23b CHO2D11) 41 2, ; ;,,, : ; ; ; SSR : S : A : X (2007) 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 , China; 2 Institute of Forestry and Pom ology, B eijing A cadem y of A griculture and Forestry Sciences, B eijing , 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 , and the genetic diversity index was 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, : ; : 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,

2 , ( 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 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 Jinfeng Xingcheng, L iaoning Hongnahe Tulufan, Xinjiang Jinhua Jinchuan, Sichuan Zaoshujuju Kuche, Xinjiang Yali Hebei P. comm unis L. Bartlett B ritain Chuizhi Yali Shijiazhuang, Hebei Flem ish BeautyBelgium Jinxian Dayali Jinxian, Hebei Clapp Favorite America P. ussuriensis Jianba L iaoning Yali Bartlett W ujiuxiang Xingcheng, L iaoning Maxim Huagai L iaoning Nanguo Bartlett Jinxiang Xingcheng, L iaoning Manyuanxiang L iaoning Kuerle xiang Xuehua Yuluxiang Taiguo, Shanxi Q iuzi Beizhen, L iaoning Pingguoli M ishirazu Zaosu Xingcheng, Liaoning Balixiang L iaoning Clappπs Favorite Zaosu Bayuehong Meixian, Shaanxi Daxiangshui L iaoning Xuehua Shinseiki Huangguan Shijiazhuang, Hebei 21 Nanguo Anshan, Liaoning

3 2 : SSR 307 CTAB DNA, 1, DNA SSR ( Gianfranceschi et al., 1998; Yamamoto et al., 2001, 2002a, 2002b) Sangon SSR : 20 L ng DNA, 10 buffer 210 L, MgCl mmol/l, dntp 012 mmol/l, 015 mol/l, Taq 1 U PCR PE PTC100 PCR 94 4 m in; 94 1 m in, m in, 72 2 m in, 10, 015 ; 94 1 m in, 1 m in, 72 2 m in, 30 ; 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 ) 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: , ( 2) DNA,,, SSR, BGT23b 41 15, 30 SSR, BGT23b CHO2D11, 41 12, pbr322 DNA /M SP1 markers SSR 41 Table 2 Amplif ied results of 41 pear cultivars with 12 SSR primers Primer Annealing ( ) Number of alleles detected h Locus heterozygosity KU KA4b KA KA KB BGT23b NH004a NH015a NH009b NH013a NH017a CHO2D Average

4 ,, ;, ,,,,, , , D ice UPGMA ( 2) , ,, ;,, ( ) ( ), ( ),, 3 F ig D endrogram of pear cultivars ba sed on SSR da ta The same code as in Table SSR ( ) ( ), Yamamoto (2001) ( O sa Gold

5 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,, , , , ;,, ,,, (, 1963; Teng et al., 2001), 3, ,,, Teng (2001) 3, (, 2001), 415% (, 1983;, 1985;, 1986) NH015a NH015a CHO2D11 3, ,,, SSR 20,,, SSR,,, References Bailey L H Standard cyclopedia of horticulture. New York: Macm illan: Chen Rui2yang, L i Xiu2lan, TongDe2yao, L iu Yu2xin, W u Q iu2qin, W ang Feng2shan, Pu Fu2shen, Huang L i2sen Study on the chromo2 some number of pear ( Pyrus L. ) in China. Acta Horticulturae Sinica, 10 (1) : ( in Chinese) 1,,,,,,, , 10 (1) : Gianfranceschi L, Seglias N, Tarchini R, Kom janc M, Gessler C Simp le sequence repeats for the genetic analysis of app le. Theor. Appl. Genet., 96: Huang L i2sen, L i Shu2ling, Pu Fu2shen Study on the chromosome number of pear cultivars in China. China Fruit, (1) : ( in Chinese)

6 310 34,, , (1) : Kikuchi A A ssessment for Chinese pear species and cultivars. Collected Records for Horticultural Research, 3: ( in Japanese) , 3: Pu Fu2shen, Huang L i2sen, Sun B ing2jun, L i Shu2ling Study on the chromosome number of wild and cultivated pears ( Pyrus sp. ) in China. Acta Horticulturae Sinica, 12 (3) : ( in Chinese),,, , 12 (3) : Pu Fu2shen, Wang Yu2lin China fruit flora, volume (pear). Shanghai: Shanghai Scientific and Technical Press: 304. ( in Chinese), g g. : : 304. Teng Y, Tanabe K Reconsideration on the origin of cultivated pears native to East A sia. Acta Horticulturae, 634: Teng Y, Tanabe K, Tamura F, Itai A Genetic relationship s of pear cultivars in Xinjiang, China as measured by RAPD markers. J. Hort. Sci. B iotech., 76: Yamamoto T, Kimura T, Sawamura Y, Kotobuki K, Ban Y, Hayashi T, Matsuta N SSR s isolated from app le can identify polymorphism and genetic diversity in pear. Theor. App l. Genet., 102: Yamamoto T, Kimura T, Sawamura Y, Manabe T, Kotobuki K, Hayashi T, Ban Y, Matsuta N. 2002a. Simp le sequence repeats for genetic anal2 ysis in pear Euphytica, 124: Yamamoto T, Kimura T, Shod AaM, Ban Y, Hayashi T, Ban Y, Matsuta N. 2002b. Development ofm icrosatellite markers in the Japanese pear ( P. pyrifolia Nakai). Molecular Ecology Notes, 2: Yang Ji The formation and evolution of polyploidy genomes in plants. Acta Phytotaxonom ica Sinica, 39 (4) : ( in Chinese) , 39 (4) : News CNKI ( ) 1.,,, /1982 / , /1991 / ,,,, /1989 / RAPD,,,, /1998 / , /1989 / AFLP,,,, /2000 / RAPD,, /1998 / ,, /1994 / ,,,, /1996 / RAPD,,,, /1998 / RAPD,, /1999 / RAPD,,, /1998 / ABA,,,, /1998 / ,,,, /1998 / ,, /1987 / , /1990 / , /1998 / ,,, /1998 / ,, /1994 / ,,,, /1997 /03 70 ( 402 )