Department of Forest Genetics, Forestry and Forest Products Research Institute, Tsukuba, Ibaraki, Japan.
Am J Bot. 2012 Jan;99(1):e28-32. doi: 10.3732/ajb.1100299. Epub 2011 Dec 27.
Robust markers are required (inter alia) for assessing origins of Calophyllum inophyllum populations on the Bonin Islands, Japan. Therefore, informative expressed sequence tag (EST)-based microsatellite or simple sequence repeat (SSRs) markers in the species were sought.
Using 135378 ESTs derived from de novo pyrosequencing, primers for 475 EST-SSRs were developed, 48 of which were tested for PCR amplification. Thirty-six of the 48 primers showed clear amplification, with 23 displaying polymorphism in sampled populations. Expected heterozygosity in the samples from the Bonin Islands and Ryukyu Islands populations ranged from 0.041 to 0.697 and from 0.041 to 0.773, respectively.
As EST-SSRs are potentially tightly linked with functional genes, and reportedly more transferable to related species than anonymous genomic SSRs, the developed primers have utility for future studies of the origins, genetic structure, and conservation of C. inophyllum and related species.
需要稳健的标记物(除其他外)来评估日本小笠原群岛上的厚叶红厚壳种群的起源。因此,在该物种中寻找基于表达序列标签(EST)的信息丰富的微卫星或简单重复序列(SSR)标记物。
使用从头焦磷酸测序获得的 135378 个 EST,开发了 475 个 EST-SSR 的引物,其中 48 个进行了 PCR 扩增测试。在采样的小笠原群岛和琉球群岛种群中,有 48 个引物中的 36 个显示出清晰的扩增,其中 23 个显示出多态性。小笠原群岛和琉球群岛种群的样本中的预期杂合度范围分别为 0.041 至 0.697 和 0.041 至 0.773。
由于 EST-SSR 可能与功能基因紧密连锁,并且据报道比匿名基因组 SSR 更能转移到相关物种,因此开发的引物可用于未来对厚叶红厚壳及其相关物种的起源、遗传结构和保护的研究。