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中国农学通报

中国农学通报 ›› 2016, Vol. 32 ›› Issue (36): 104-108.doi: 10.11924/j.issn.1000-6850.casb16050162

所属专题： 园艺

• 生物技术科学 • 上一篇    下一篇

甜菜EST-SSR引物的快速筛选

吴则东1,2,3,邹兰兰4,刘乃新1,2,3,马龙彪1,2,3,倪洪涛4   

  1. (1黑龙江省普通高校甜菜遗传育种重点试验室/黑龙江大学,哈尔滨 150080；2中国农业科学院甜菜研究所/黑龙江大学农作物研究院,哈尔滨 150080；3中国农业科学院北方糖料作物资源与利用重点开放试验室,哈尔滨 150080;4黑龙江大学农业资源与环境学院,哈尔滨 150080）
  • 收稿日期:2016-05-27 修回日期:2016-12-15 接受日期:2016-08-25 出版日期:2016-12-26 发布日期:2016-12-26
  • 通讯作者: 倪洪涛
  • 基金资助:
    甜菜现代产业技术体系建设项目东北区育种岗位专家(CARS-210104)。

A Rapid Screening of EST-SSR Primers in Beet

Wu Zedong1,2,3, Zou Lanlan4, Liu Naixin1,2,3, Ma Longbiao1,2,3, Ni Hongtao4   

  1. (1Key Laboratory of Sugar Beet Genetic Breeding/Heilongjiang University, Harbin 150080; 2Sugar Beet Research Institute of Chinese Academy of Agricultural Sciences/Crop Academy of Heilongjiang University, Harbin 150080; 3Key Laboratory of North Sugar Crop Resource and Utilization, Chinese Academy of Agricultural Sciences, Harbin 150080; 4College of Agricultural Resources and Environment Science, Heilongjiang University, Harbin 150080)
  • Received:2016-05-27 Revised:2016-12-15 Accepted:2016-08-25 Online:2016-12-26 Published:2016-12-26

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摘要： 为了快速从甜菜EST-SSR引物中筛选出适合甜菜分子生物学的引物，利用12个不同的甜菜品种对80对甜菜EST-SSR引物进行扩增，同时设置12个不同的退火温度，利用8%的非变性聚丙烯酰胺凝胶对PCR产物进行分离。结果表明：80对EST-SSR引物虽然均能够扩增出产物，但绝大部分引物扩增的条带由于没有多态性或者条带难以辨别而不能够使用，只有13对引物扩增出了清晰、易于识别的条带，有4对引物多态性较高，其余9对引物多态性均较低，而梯度退火温度则表明，退火温度对EST-SSR引物影响不大，只有个别引物会在较低退火温度条件下产生非特异性的条带，因此将所有引物的退火温度均设定为60℃。利用不同品种以及梯度退火温度可以实现EST-SSR引物的快速筛选，但要想筛选出多态性高的引物还需要对更多的EST-SSR引物进行筛选。

关键词: 牛, 牛, 体外成熟, 体细胞核移植, 化学激活, 胚胎培养

Abstract: To rapidly screen out the primers suitable for the molecular biology of sugar beet from EST-SSR primers of sugar beet, 12 varieties of sugar beet were used to amplify with 80 pairs of EST-SSR primers of sugar beet. At the same time, 12 kinds of annealing temperatures were set and 8% non-denaturing PAGE was used to separate the PCR products. The results showed that although all 80 pairs of EST-SSR primers could amplify products, most of them were useless because they were not polymorphic or difficult to distinguish. There were only 13 pairs of primers amplifying bands which were clear and easy to distinguish. Among them, 4 pairs had high polymorphism and the other 9 pairs had low polymorphism. While the gradient annealing temperature indicated that the annealing temperature had no obvious influence on EST-SSR primers and only several primers produced unspecific bands under low annealing temperature. So the annealing temperature of all primers was set to be 60 ℃. Quick screening of EST-SSR primers could be achieved using different varieties and gradient annealing temperature. But more EST-SSR primers should be screened to get primers with high polymorphism.

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