甜菜EST-SSR引物的开发与应用  

Development and Utilization of EST-SSR Marker in Sugarbeet(Beta vulgaris L.)

作　　者：史树德[1] 魏磊[1] 张子义[2] 邵金旺[3] 田自华[1] 

机构地区：[1]内蒙古农业大学农学院,呼和浩特010019 [2]内蒙古农业大学生命科学院,呼和浩特010018 [3]内蒙古农业大学甜菜生理研究所,呼和浩特010018

出　　处：《中国糖料》2011年第3期1-5,共5页Sugar Crops of China

基　　金：国家甜菜产业技术体系(CARS-210402);内蒙古自治区高等学校科学研究项目(NJzy08045)

摘　　要：利用NCBI公共数据库现有的甜菜(Beta vulgaris L.)表达序列标签(expressed sequence tags,EST)数据信息,开发了甜菜EST-SSR标记。在所有的29830条甜菜EST序列中共确认得到20109条非冗余EST序列,总长为11287.6kb。在含有微卫星重复的6951条EST序列中按照SSR引物设计要求,最终获得了2845个EST-SSR,平均每3.96 kb含有1个SSR。EST-SSR的分布频率和特征分析表明,A/T单碱基重复最多,其次是AAG/CTT三核苷酸重复,AG/CT二核苷酸重复,ACCTCC/AGGTGG等六核苷酸重复最少。随机合成了100对SSR引物,并分别选用6个甜菜品种进行多态性检验,将其按遗传相似性分为两组,多态信息含量(polymorphism information content,PIC)平均值为0.47。本研究证实这种全新的开发甜菜SSR标记的方法具有高效、多态性较高的特点,在甜菜遗传多样性分析、功能基因定位、遗传图谱构建以及比较基因组等研究方面有广阔的利用前景。The increasing availability of expressed sequence tags（ESTs） sequences provides an invaluable resource of EST-SSR markers.The EST sequences encoding transcription used in this study included 6951 ESTs with SSR repeats downloaded from NCBI databases.A total of 4338 microsatellites（SSRs） were isolated from the 6951 sugarbeet expressed sequence tags（ESTs）.According to primer design criteria,2845 primer pairs for EST-SSR were designed.Under an appropriate polymerase chain reaction（PCR） system,randomly synthesized 100 EST-SSR primer pairs were screened against genomic DNA of 6 sugarbeet varieties,and 47 EST-SSR primer pairs out of the above 100 resulted in PCR products.Then,all 47 pairs were detected in six sugarbeet varieties for polymorphisms,three native and three foreign cultivars were clustered two groups,and 25 pairs（53.2%） were found to be polymorphic,accounting for 0.97% of the total number of designed primer pairs.Those results demonstrate the possibility of developing EST-SSR markers using sugarbeet EST-SSR markers.

关 键 词：甜菜 微卫星 表达序列标签 遗传多样性 

分 类 号：S566.303[农业科学—作物学] Q78[生物学—分子生物学]