Development and utility of EST-SSR markers in Ulva prolifera of the South Yellow Sea
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摘要: 石莼属绿藻由于海水富营养化大量增殖形成绿潮以及海洋污染,有关绿潮生物大规模增殖爆发的原因和机制亟待解决。因此,本研究采用EST-SSR分子标记对中国南黄海几个浒苔群体进行遗传多样性分析。从8179条石莼科EST序列(新测定6203条,NCBI数据库下载1976条)中共搜索到238个微卫星位点,设计出37对引物并从中成功筛选出11对引物对南黄海69份近岸浒苔样品和13份漂流样品进行遗传多样性分析。引物通用性分析结果表明,有6对引物在石莼属其它5个物种均有良好的扩增,平均扩增位点4.67个。UPGMA聚类分析结果表明:所有82份材料能够明显的区分开来,来自同一采样地的样品大多数都能够按照各自所在群体聚为一支,显示出良好的地理相关性。其中13份漂流样品区别于其它近岸样品独自聚为一支(HT-001-02除外)。
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关键词:
- 浒苔 /
- 表达序列标签(EST) /
- 微卫星 /
- 遗传多样性 /
- 海藻 /
- 简单序列重复(SSR)
Abstract: Ulva species can grow rapidly in nutrient-rich habitats causing green tides and marine fouling. A more complete understanding of the reasons behind these outbreaks is urgently required. Accordingly, this study attempts to use microsatellite markers based expressed sequence tag (EST) to analyze the genetic variation of several Ulva prolifera populations in the South Yellow Sea of China. Two hundred and thirty-eight SSRs were identified from 8 179 unique ESTs (6 203 newly sequenced and 1 976 downloaded from NCBI database) and 37 primer pairs were successfully designed according to the ESTs; 11 pairs were selected to detect the genetic diversity and relationship of 69 attached U. prolifera samples and 13 free-floating samples collected from coastal and off-coast areas of the South Yellow Sea. The results of cross-species transferability showed that six of the 11 EST-SSR primers could give good amplification in other five Ulva species and the average allele number was 4.67. Genetic variation analysis indicated that all 82 U. prolifera samples were clearly divided and most samples collected from the same site clustered together as a group in the dendrogram tree produced by unweighted pair-group mean analysis (UPGMA) method and the cluster results showed some consistency with the geographical origins. In addition, 13 free-floating samples (except HT-001-2) were grouped as a single clade separated from the attached samples. -
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