Lee G P, Lee C H, Kim C S
Institute of Natural Science, Seoul Women's University, 139-240 Seoul, Korea.
Theor Appl Genet. 2004 May;108(8):1487-91. doi: 10.1007/s00122-003-1582-8. Epub 2004 Jan 29.
We generated RAPD, SCAR, and conserved 18S rDNA markers for classifying and identifying cultivars of Pyrus pyrifolia (Japanese pear) and P. communis (European pear). PCR amplification with selected specific primers-LCH327UP and LCH327DOWN-was performed using DNA extracted from 25 P. pyrifolia and P. communis cultivars. The 1,380-bp fragment was amplified from P. communis cvs. Beurre Giffard, Cascade, Conference, Clapp's Favorite, Packhams Triumph, and Winter Nelis. RAPD has only a dominant single band of 1,380-bp, however, SCAR has one or more band of the same size. Amplification involving sequence-specific primer pairs LCH346UP and LCH346DOWN resulted in a loss of polymorphism. The 1,190-bp fragment was amplified from all P. pyrifolia cultivars. The conserved sequences of the 18S rDNA fragment of 25 pear cultivars were amplified and analyzed with 42 restriction enzymes. Compared with P. pyrifolia cultivars, they lacked the restriction enzyme site of KpnI and had one less RsaI site. Cultivar Gamcheonbae had a specific PstI restriction site, while cvs. Mansoo and Conference pear digested with AluI showed a different presentation than other cultivars. For the Okusankichi and Shinil pears TaqI was best marker for identification in P. pyrifolia. These results can be adopted for identifying pear cultivars; to date there is no standard marker for identifying the cultivars of fruit trees in Korean fruit tree breeding programs.
我们生成了随机扩增多态性DNA(RAPD)、序列特异性扩增区域(SCAR)和保守的18S核糖体DNA(rDNA)标记,用于对沙梨(日本梨)和西洋梨(欧洲梨)品种进行分类和鉴定。使用从25个沙梨和西洋梨品种中提取的DNA,用选定的特异性引物LCH327UP和LCH327DOWN进行聚合酶链反应(PCR)扩增。从西洋梨品种白来发、卡司开德、康佛伦斯、克拉普最爱、帕克斯胜利和冬梨中扩增出了1380碱基对的片段。然而,RAPD只有一条1380碱基对的显性单带,而SCAR有一条或多条相同大小的带。涉及序列特异性引物对LCH346UP和LCH346DOWN的扩增导致多态性丧失。从所有沙梨品种中扩增出了1190碱基对的片段。用42种限制性内切酶对25个梨品种的18S rDNA片段的保守序列进行了扩增和分析。与沙梨品种相比,它们缺乏KpnI的限制性酶切位点,且RsaI位点少一个。品种甘泉梨有一个特定的PstI限制性酶切位点,而用AluI消化的万秀梨和康佛伦斯梨与其他品种呈现出不同的图谱。对于奥山金吉梨和新日梨,TaqI是沙梨品种鉴定的最佳标记。这些结果可用于梨品种的鉴定;在韩国果树育种计划中,目前尚无用于鉴定果树品种的标准标记。