甜樱桃SSR体系的建立、优化及应用Establishment,optimization and application of SSR system in sweet cherry
张琪静;张新忠;李贺;谷大军;李春敏;张志宏;
摘要(Abstract)：
为摸索适宜甜樱桃的SSR反应体系,利用正交设计对Mg2+、dNTPs、引物、Taq酶和模板DNA等5种因素4个水平进行筛选和优化,20μL反应体系中,Mg2+、dNTPs和引物的最适浓度为1.5mmol/L、0.2mmol/L、0.4μmol/L,Taq酶宜加入1U,模板DNA应加入10～40ng;引物的最佳退火温度为56.0～62.8℃。用10对引物及建成的反应体系对10个供试品种扩增,6%的非变性聚丙烯酰胺凝胶电泳检测,品种间DNA谱带多态性丰富,共有29个等位位点,证实该体系稳定可靠。
关键词(KeyWords)： 甜樱桃;正交试验;SSR;反应体系
基金项目(Foundation):
作者(Author): 张琪静;张新忠;李贺;谷大军;李春敏;张志宏;
Email:
参考文献(References)：
- [1]WEBSTER A D.The taxonomic classification of sweet and sour cherries and a brief history of their cultivation[M]//In:Webster A D,Looney N E(eds)Cherries:Crop physiology,production and uses,CAB.International:Wallingford,Oxon,UK,1996:3-24.
- [2]MARTINEZ-GOMEZ P,ARULSEKAR S,POTTER D,DEJONG T,GRADZIEL T M,BLISS F A.SSR variation within and between peach and almond cultivars[M].San Diego,California,2000:541.
- [3]ARANZANA MJ,GARCIA-MAS J,CARBO J,ARUS P.Develop-ment and variability analysis of microsatellite markers in peach[J].Plant Breed,2002,121:87-92.
- [4]CIPRIANI G,LOT G,HUANG W-G,MARRAZZO MT,PETER-LUNGER E,TESTOLINR.AC/GTand AG/CT microsatellite repeats in peach[Prunus persica(L)Batsch]:isolation,characterization and cross-species amplification in Prunus[J].Theor Appl Genet,1999,99:65-72.
- [5]DEVROOCQ V,FAV M G,HAGEN L,BORDENAVE L,DE-CROOCQ S D.Evelopment and transferability of apricot and grape ESTmicrosatellite markers across taxa[J].Theor Appl Genet,2003,106:912-922.
- [6]SOSINSKI B,GANNAVARAPU M,HAGER L D,BECK L E,KING G J,RYDER C D,RAJAPAKSE S,BAIRD W V,BAL-LARD R E,ABBOTT A G.Characterization of microsatellite mark-ersin peach[Prunus persica(L.)Batsch][J].Theor Appl Genet,2000,101:421-428.
- [7]STUSS D,AHMAD R,SOUTHWICK S M.Analysis of sweet cherry cultivars(Prunus avium L.)using SSR and AFLP markers[J].J Amer Soc Hort Sci,2003,128:904-909.
- [8]TESTOLIN R,MARRAZZO T,CIPRIANI G,QUARTA R,VERDE I,DETTORI M,PANCALDI M,SANSAVINI S.Microsatellite DNA in peach[Prunus persica(L.)Batsch]and its use in fingerprintingand testingthegeneticoriginofcultivars[J].Genome,2000,43:512-520.
- [9]YU ML,MA R J,XU J L,SHEN Z J,ZHANG Z.Identification of genetic relationship of peach species by SSR[J].Journal of Fruit Sci-ence,2004,21:106-112.(in Chinese)俞明亮,马瑞娟,许建兰,沈志军,章镇.桃种间亲缘关系的鉴定[J].果树学报,2004,21:106-112.
- [10]XU X X,LIANG H Y,ZHEN Z X,YANG M S.Establishment of SSR analysis systemin apple[J].Journal of Fruit Science,2006,23(2):161-164.(in Chinese)徐兴兴,梁海永,甄志先,杨敏生.苹果SSR反应体系的建立[J].果树学报,2006,23(2):161-164.
- [11]REDDY MP,SARIL N,SIDDIQ E A.Inter simple sequence repeat(ISSR)polymorphism and its application in plant breeding[J].Eu-phytica,2002,128(1):9-17.
- [12]TANG Q Y,FENG M G.PS data processing system for practical statistics[M].Beijing:Science Press,2002:147-148.(in Chinese)唐启义,冯明光.实用统计分析及其DPS数据处理系统[M].北京:科学出版社,2002:147-148.