口虾蛄转录组微卫星标记开发

Development of Microsatellite Markers in Oratosquilla oratoria Transcriptome

采用高通量测序技术获得的口虾蛄转录组测序数据,筛选具有多态性的SSR标记。结果共获得87355条Unigene,67657个微卫星位点,其中双碱基重复和三碱基重复的SSR较多,分别有30235个和17590个,占总数的44.69%和26%。获得230种重复类型,其中双碱基中的AC/GT重复和三碱基中的AGG/CCT重复是重复次数较多的重复类型。从中挑选碱基重复次数较多的微卫星引物共50对进行合成。用北海24个口虾蛄样品进行验证,获得条带清晰和多态性好的引物6对。6个微卫星位点在口虾蛄北海群体的遗传多样性分析中均表现出较高的多态性。平均有效等位基因数(k)为5.17,观测杂合度(Ho)为0.4372,平均期望杂合度(He)为0.6724,多态信息含量为(PIC)为0.6023,这说明开发的6对SSR标记均适合群体遗传多样性研究。为口虾蛄进一步遗传多样性的研究奠定了基础。

This paper, we use high-throughput sequencing technology to obtain the sequencing data of the Oratosquilla oratoria transcriptome and filter and develop polymorphic SSR markers. A total of 87 355 unigene were obtained, include 67 657 microsatellite loci, which mostly SSRs were Di-nucleotide repeats and Tri-nucleotide repeats, with 30 235 and 17 590, respectively. A total of 230 types of repetition were obtained. In Di-nucleotide SSR repeats, the most repeated type was AC/GT repetition In the Tri-nucleotide repeats of SSR, the repeated dominant type is AGG/CCT repetition. The 50 pairs of microsatellite primers with high number of base repetitions were selected to synthesize. A total of 6 pairs of SSR markers with clear bands and polymorphisms were identified by using 24 samples from Beihai. 6 microsatellite loci showed high polymorphism in the genetic diversity analysis of Beihai population.The average effective number of allele was 5.17, expected heterozygosities and Average observed (He and Ho) were 0.437 2 and 0.672 4. Average polymorphism information content (PIC) is 0.602 3, result showed all of the 6 pairs of SSR markers were suitable for population genetic diversity. It lays a foundation for the further study of genetic diversity of O. oratoria.