栽培种花生EST-SSR引物的开发及应用  

Development and Utilization of EST-SSR Primers in Peanut(Arachis hypogaea L.)

作　　者：肖洋[1] 廖伯寿[1] 晏立英[1] 黄家权[1] 任小平[1] 雷永[1] 姜慧芳[1] 

机构地区：[1]中国农业科学院油料作物研究所/油料作物生物学重点开放实验室,武汉430062

出　　处：《湖北农业科学》2010年第11期2625-2628,共4页Hubei Agricultural Sciences

基　　金：国家"863"计划项目(2006AA10A115;2007AA100106);国家自然科学基金项目(30871626)

摘　　要：从高油抗青枯病花生新品系"06-4104"构建的5个cDNA文库获得了63207条EST,经序列拼接,获得14547条Uni-EST。经MISA检索,在这些Uni-EST中共检测出2643个SSR位点,分布于2250条EST中,发生频率为15.5%,平均每3.1kbEST序列含有一个SSR位点。其中,三核苷酸重复类型出现频率最高,占SSR总数的49.1%,其次是二核苷酸重复类型,占SSR总数的45.8%。在发现的46类重复基序中,AG/TC重复基序的频率最高,AAG/TTC次之。利用Primer3从含有SSR的2250条EST中共设计引物100对,利用这些引物检测花生栽培品种的多态性。结果表明,在所设计的100对引物中有91对在供试的6个花生栽培品种中得到有效扩增,其中13对得到了多态性的扩增产物,每对引物检测出的等位基因数2～3个,平均2.2个。A total of 63 207 ESTs were produced from 5 cDNA libraries of RNAs of 06-4104,a peanut（Arachis hypogaea L.） line with high oil content and resistance to bacterial wilt.A total of 2 643 SSRs were identified,distributed in 2 250 ESTs.On average,one SSR was contained in 3.1 kb EST sequence with tri-nucleotide motifs（49.1%） as the most abundant motif types followed by di-nucleotide motifs（45.8%）.AG/TC motifs＇ frequency is highest with AAG/TTC the next in 46 motifs found.Based on the 2 250 SSR-EST,100 primer pairs were designed and synthesized to test the polymorphisms of in 6 peanut cultivars.The results showed that 91 primer pairs could be amplified effectively in the peanut cultivars,13 primer pairs of which exhibited polymorphisms with 2～3 alleles,an average of 2.2 alleles each.

关 键 词：花生 EST-SSR 标记开发 

分 类 号：S565.2[农业科学—作物学] Q789[生物学—分子生物学]