仿刺参的微卫星标记

Microsatellite markers of Apostichopus japonicus

为了评价种质资源及基础生物学研究的需要，本文开发了仿刺参的微卫星标记。NCBI数据库中共有20个含有仿刺参微卫星的序列，从中选取8个设计引物，发现6个微卫星位点有多态性。不同的引物获得的等位基因数为3～9个不等，6个位点共获得了31个等位基因，每个位点平均获得5．2个等位基因。6个位点的平均观测杂合度（Ho）为0．3611，平均期望杂合度（He）为0．6402。位点AJMS004提供的多态性信息含量值较低，为0．4862；其他5个位点均在0．5以上。另外，还尝试了红海参（Parastichopus californicus）微卫星标记在仿刺参的通用性。实验结果表明，在较高的退火温度下，5对引物均能扩增仿刺参的基因组DNA并具多态性。5个位点共获得了22个等位基因，每个位点平均获得4．4个等位基因。5个位点的平均观测杂合度（Ho）为0．1733，平均期望杂合度（He）为0．4201。其中位点Psc2的多态性信息含量值最高，为0．8500。

Sea cucumber （Apostichopus japonicus ） is regarded as a tonic food and traditional medicine in China. In order to aid in investigation of the population structure and marker assisted breeding, 11 microsatellite markers were developed and evaluated with sixty individuals caught from the offing in Shandong Province, China. In the NCBI database, 20 sequences of Apostichopus japonicus were found to contain microsatellites when data mining was performed. Eight microsatellite-containing sequences were chosen for designing the PCR primers. The results showed that six primer pairs successfully amplified scorable PCR products and revealed polymorphism with 5.2 alleles per locus. The locus AJMS007 had the greatest number of alleles （9）, while the locus AJMS004 had the lowest number of alleles （3）. The average Ho and He values of six loci were 0.3611 and 0.6402, respectively. PIC value was 0.4862 for locus AJMS004, and more than 0.5 for the other five loci. Five microsatellite markers developed and characterized in Parastichopus californicu were used to assess for transferability analysis in Apostichopus joponicus. The results showed that all the five primer pairs could amplify clear and scorable PCR products with expected size in Apostichopus japonicus. A total of 22 alleles were obtained for the five loci, with 4.4 alleles per locus. The locus Psc 2 had the greatest number of alleles （9） and the locus Psc 3 had the lowest number of alleles （2）. The average Ho and He values of the five transferred loci were 0. 1733 and 0.4201, respectively. And Psc 2 showed the highest PIC value of 0.8500.