Development of microsatellite markers in Primula sieboldii E. Morren, a threatened Japanese perennial herb
Development of microsatellite markers in Primula sieboldii E. Morren, a threatened Japanese perennial herb
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DOI：
10.1023/b:coge.0000006121.82993.5f
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发表时间：
2003-11
影响因子：
2.2
通讯作者：
S. Ueno;Y. Tsumura;I. Washitani
中科院分区：
文献类型：
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作者：
S. Ueno;Y. Tsumura;I. Washitani
微卫星标记越来越多地应用于许多研究领域，例如基因组作图、群体遗传学、法医学和受威胁物种保护（Goldstein 和 Schlotterer 1999）。微卫星的高度多态性使其适合识别克隆，以及分析繁殖系统和亲缘关系，从而建立保护生物学的关键参数。报春花是一种多年生草本植物，生长在日本的一系列潮湿生境中。尽管西氏松曾经很常见，但它已经濒临灭绝，现在被日本环境厅（1997 年）列入日本植物红色名录中的易危物种。威胁它的主要过程包括园艺采集、自然演替和房地产开发造成的栖息地丧失（环境署 1997 年）。 Isagi等人(2001)首先开发了该物种的微卫星标记，但他们开发的微卫星中有一半以上显示出低水平的多态性，在他们的20个样本中只有两到三个等位基因。在这里，我们描述了七个高度多态性的微卫星标记，因此对于自然群体的遗传分析更有价值。通过改良的 CTAB 方法（Murray 和 Thompson 1980）提取西氏假单胞菌的基因组 DNA，并通过超速离心纯化。 DNA 样品 (10 µg) 用 NdeII 消化并在 1.2% 琼脂糖凝胶上进行电泳分离。回收了 300 至 1000 bp 的 DNA 片段。将片段（约 600 ng）连接至提供的 Sau3AI 盒（5 pmol）
Microsatellite markers are being increasingly used in many areas of research, such as genome mapping, population genetics, forensic science and conservation of threatened species (Goldstein and Schlotterer 1999). The highly polymorphic nature of microsatellites makes them suitable for identifying clones, as well as for analyzing breeding systems and kinship, and thus establishing critical parameters for conservation biology.Primula sieboldii E. Morren is a perennial herb that occurs in a range of moist habitats in Japan. Although P. sieboldii was once very common, it has become endangered and is now listed as Vulnerable in the Japanese plant red list by the Japanese Environment Agency (1997). The major processes threatening it include horticultural collection, natural succession and habitat loss through property development (Environment Agency 1997). Microsatellite markers for this species were first developed by Isagi et al.(2001), but more than half of the microsatellites they developed displayed low levels of polymorphism, with only two or three alleles among their 20 samples. Here, we describe seven microsatellte markers that are highly polymorphic and therefore more valuable for genetic analysis in natural populations. Genomic DNA of P. sieboldii was extracted by a modified CTAB method (Murray and Thompson 1980) and purified by ultracentrifugation. Samples of DNA (10 µg) were digested with NdeII and electrophoretically separated on a 1.2% agarose gel. DNA fragments ranging from 300 to 1000 bp were recovered. The fragments (approximately 600 ng) were ligated to Sau3AI cassette (5 pmol) supplied