首页 | 本学科首页   官方微博 | 高级检索  
     检索      

甜樱桃SSR体系的建立、优化及应用
引用本文:张琪静,张新忠,李贺,谷大军,李春敏,张志宏.甜樱桃SSR体系的建立、优化及应用[J].果树学报,2007,24(6):770-773.
作者姓名:张琪静  张新忠  李贺  谷大军  李春敏  张志宏
作者单位:1. 沈阳农业大学园艺学院,沈阳,110161;辽宁省果树科学研究所,辽宁熊岳,115214
2. 河北省农林科学院昌黎果树研究所,河北昌黎,066600
3. 沈阳农业大学园艺学院,沈阳,110161
4. 辽宁省果树科学研究所,辽宁熊岳,115214
摘    要: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:甜樱桃  正交试验  SSR  反应体系
文章编号:1009-9980(2007)06-770-04
修稿时间:2007-05-15

Establishment,optimization and application of SSR system in sweet cherry
ZHANG Qi-jing,ZHANG Xin-zhong,LI He,GU Da-jun,LI Chun-min,ZHANG Zhi-hong.Establishment,optimization and application of SSR system in sweet cherry[J].Journal of Fruit Science,2007,24(6):770-773.
Authors:ZHANG Qi-jing  ZHANG Xin-zhong  LI He  GU Da-jun  LI Chun-min  ZHANG Zhi-hong
Institution:1 College of Horticulture ,Shenyong Agrieultural University, Shenyang, Liaoning 110161 China; 2 Liaoning Institute of Pomology, Xiongyue, Liaoning 115214 China; 3 Chongli Fruit Research Institute, Hebei Academy of Agricultural and Forestry Sciences ,Chongli, Hebei 066600 China
Abstract:Using orthogonal design,five factors including Mg2+,dNTPs,primer,Taq polymerase as well as template DNA at four content levels were screened and optimized in order to establish SSR reaction system in sweet cherry. The results showed that 1.5 mmol/L Mg2+,0.2 mmol/L dNTPs,0.4 μmol/L primer,1 U Taq polymerase,10 to 40 ng template DNA were the optimal conditions in the reaction of 20 μL volume. The optimal annealing temperature ranged from 56.0 ℃ to 62.8 ℃. Under optimal conditions,SSR reactions were carried out with 10 cultivars of sweet cherry using 10 pairs of primers. PCR products were tested by electrophoresis using 6% nondenatured polyacrylamide gels. The results revealed high polymorphism with 29 alleles in whole which demonstrated that the reaction system established in this study was stable and reliable.
Keywords:Sweet cherry  Orthogonal design  SSR  Analysis system
本文献已被 CNKI 维普 万方数据 等数据库收录!
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号