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利用菜薹转录组分析检测到48 975条unigene(38.17 Mb)序列数据,运用MIcro SAtellite(MISA)工具发现其中具有1~6个核苷酸重复类型的SSR位点11 879个,SSR位点发生频率为1/3.2 kb(312.5/Mb)。6种SSR位点类型中主要为1~3个核苷酸重复,占总SSR位点数的99.01%,其中单、二和三核苷酸类型分别为41.11%、28.23%和29.67%。发现重复序列的基序58个,重复次数在5~23之间,其中出现频率高的基序主要有A/T、AG/CT、AT/AT、AC/GT、AAG/CTT和AGG/CCT。重复序列长度在10~60 bp之间,大多小于20 bp,大于20 bp的仅有7.9%(938个SSR位点)。设计出676对具有潜在多态性的SSR引物组合,从中随机抽取30对引物进行PCR扩增验证其可用性,发现22对引物在4份菜薹种质中的扩增产物条带清晰稳定,其中12对引物具有多态性。
A total of 48 975 unigene (38.17 Mb) sequence data were detected by flowering transcriptome analysis. 11 879 SSR loci with 1 ~ 6 nucleotide repeat types were detected by MIcro SAtellite The frequency is 1 / 3.2 kb (312.5 / Mb). The six SSR loci were mainly 1 to 3 nucleotide repeats, accounting for 99.01% of the total SSR loci, of which the single, double and triple nucleotide types were 41.11%, 28.23% and 29.67%, respectively. The motifs of repeat sequences were found to be 58 and the number of repeats was between 5 and 23. The most frequent motifs were A / T, AG / CT, AT / AT, AC / GT, AAG / CTT and AGG / CCT . The length of the repeats was between 10 and 60 bp, mostly less than 20 bp, and only 7.9% (938 SSR sites) of> 20 bp. Sixty-six pairs of SSR primer combinations with potential polymorphism were designed. 30 pairs of primers were randomly selected for PCR amplification to verify their availability. The bands of amplification products of 22 pairs of primers were found to be clear and stable, 12 pairs of primers with polymorphism.