摘要
实验采用SAM法,以"台农17号"菠萝作为开发基因组SSR分子标记的植物材料,并对开发获得的SSR标记在16个菠萝品种中进行了验证。利用两对锚定引物和12对接头引物筛选菠萝基因组SAM文库,从中开发得到86条含有SSR位点的序列,对这86条序列用MISA软件搜索后得到94个SSR位点,其中单核苷酸重复3个,二核苷酸重复91个。二核苷酸重复仅有2种重复单元,一种是GA/CT,占二核苷酸重复总数的57.14%(52/91),另一种是AC/GT,占二核苷酸重复总数的42.86%(39/91)。研究未发现三核苷酸及以上的重复单元。36对SSR引物对16个材料的扩增结果显示,24对引物能够扩增出清晰、重复性好且符合预期大小的条带,其中13对引物具有多态性,可成为下一步构建菠萝指纹图谱、遗传多样性分析及基因定位等研究中有用的SSR标记。
We developed genomic SSR markers from a pineapple cultivar Tainong 17 using selectively amplified microsatellite （SAM） method and checked the polymorphism of these SSR markers in a collection of 16 pineapple accessions. In the study, a SAM library from Tainong 17 was screened with two anchored primers and 12 adapter primers. A total of 86 sequences were developed. And 94 SSR motifs from 86 sequences were identified using MISA software. 91 of 94 SSR motifs were dinucleotide repeats （DNRs）. The GA/TC motif was the most common among DNRs, accounting for 57.14% （52/91）, followed by AC/GT （42.86%）. The rest 3 SSR motifs were single nucleotide repeat . Motifs of the three and more nucleotide repeats were not found. Of the 36 primer pairs, 24 successfully amplified clear, reproducible and expected size products among 16 pineapple accessions. Of the 24 functional primers, 13 amplified polymorphic DNA among all the accessions, which could be the useful SSR markers for the research such as fingerprint, genetic diversity and gene location etc.
出处
《果树学报》
CAS
CSCD
北大核心
2010年第4期551-555,共5页
Journal of Fruit Science
基金
国家科技基础条件平台重点项目(2005DKA21000-5-43)
