基于转录组的白沙蒿SSR分子标记的开发

Development of SSR molecular markers for Artemisia sphaerocephala based on transcriptome

[目的]利用转录组数据开发白沙蒿的SSR分子标记,探索其分布特点并对SSR引物进行有效性检测。[方法]以9个不同样品白沙蒿转录组数据为基础,利用MISA软件对1kb以上的Unigenes进行筛选;把重复序列单元相同、重复次数存在差异、侧翼序列长度完全相同的SSR位点作为引物合成的标准,采用PCR技术和聚丙烯酰胺凝胶电泳技术对其有效性进行初步验证。[结果]在20 149条Unigenes中共检测出5 692个SSR位点,发生频率为22.95%,平均每隔6.66 kb出现1个位点。优势重复单元单、三、二核苷酸分别占总SSR位点的52.09%、28.30%和17.38%。经过验证筛选出具有多态性且扩增条带清晰的引物18对。[结论]印证了利用白沙蒿转录组测序产生的Unigene信息可作为开发SSR标记的有效来源,且开发的有效SSR引物可用于后续群体的相关研究中。

[Objective] The SSR markers of Artemisia sphaerocephala were developed by using transcriptome data,explored the distribution characteristics and detected the effectiveness of SSR primers. [Methods] Based on the transcriptome data of 9 different samples of Artemisia sphaerocephala,more than 1 kb Unigenes were screened by MISA software. The SSR loci with the same repeat units,difference of the repeat numbers and the same length of flanking sequences in the 9 samples were used as the standard for primer synthesis,and used the PCR amplification and polyacrylamide gel electrophoresis to verified its effectiveness. [Results]A total of 5 692 SSRloci were detected in 20 149 unigenes,with a frequency of 22. 95%,average distance of6. 66 kb. Mononucleotides,trinucleotides and dinucleotides were major types,accounted for 52. 09%,28. 30% and 17. 38%.After verification,18 pairs of primers with clear amplification products and polymorphic were selected. [Conclusion]It was confirmed that the Unigene information generated by the transcriptome of Artemisia sphaerocephala can be used as an effective source for development of SSR markers,and the development of effective SSR primers can be used in subsequent research.