| 引用本文: | 吴 欣,沈和定,王冬凤,刘 宸,杨铁柱,刁 亚.基于Illumina HiseqTM 2000高通量转录组测序的里氏拟石磺SSR 标记开发[J].海洋科学,2015,39(11):20-25. |
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| 摘要: |
| 为给石磺科贝类系统进化研究和资源保育工作提供更有效的分子标记, 使用Illumina HiseqTM 2000高通量测序技术对里氏拟石磺进行转录组测序, 以所得的拼接序列为基础, 利用MISA 和SSR Hunter1.3软件进行微卫星序列的查找, 对选出的51 个微卫星位点进行引物设计, 经PCR扩增得到单一清晰扩增条带的位点24个, 研究湛江里氏拟石磺群体扩增。结果表明, 其中14个位点表现多态性。14个多态性位点得到的等位基因数为2～4, 其中观测杂合度(Ho)和期望杂合度(He)分别为0.031 3～0.714 3 和0.031 3～0.708 4,多态性信息含量(PIC)为0.089 5～0.640 5, 14个位点中的2个位点表现为高度多态性(PIC>0.5), 10个位点表现为中度多态性(0.5>PIC>0.25), 另外2个位点表现为低度多态性(PIC<0.25)。经Sequential Bonferroni校正的Hardy-Weinberg平衡检验, 9个位点不偏离平衡(P>0.05)。开发的SSR标记可用于里氏拟石磺群体遗传学和石磺科贝类间的系统发生研究, 并为里氏拟石磺的群体种质保护提供数据支持。 |
| 关键词: 里氏拟石磺 转录组 微卫星标记 |
| DOI：10.11759/hykx20150803002 |
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| 基金项目:国家自然科学基金(41276157), 上海高校水产学一流学科建设项目 |
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| Development of microsatellite markers for Paraoncidium reevesii based on transcriptome sequencing Illumina HiseqTM 2000 |
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| Abstract: |
| Repeats in the transcriptome sequences of Paraoncidium reevesii were searched using MISA and SSR Hunter1.3, 51 sequences with SSR were selected for primier designing. 24 primers were produced clearly defined bands after preparatory screening. The further test in Zhanjiang population indicated that 14 loci were showed polymorphism. The allele numbers per locus ranged from 2 to 4 and the observed and expected heterozygosity ranged from 0.0313 to 0.7143 and from 0.0313 to 0.7084, respectively. The polymorphic information contents (PIC) of each locus were between 0.0895 and 0.6405. Among 14 loci, 2 had a PIC>0.5, 10 had a 0.5>PIC>0.25, 2 had a PIC<0.25. 9 loci were detected not deviated from HWE(P>0.05)in 14 after Bonferroni correction. No pairs of loci showed significant linkage disequilibrium. These loci can be used for the research of population genetic and phylogenetic analysis of Onchidiidae. Furthermore, it will provide molecular data to conservation of P. reevesii resources. |
| Key words: Paraoncidium reevesii transcriptome microsatellite marker |
