Health Innovations Research Institute, School of Applied Sciences, RMIT University, Melbourne, VIC 3000, Australia.
Mol Biotechnol. 2013 Jul;54(3):770-83. doi: 10.1007/s12033-012-9625-5.
Salvia is an important genus from the Lamiaceae with approximately 1,000 species. This genus is distributed globally and cultivated for ornamental, culinary, and medicinal uses. We report the construction of the first fingerprinting array for Salvia species enriched with polymorphic and divergent DNA sequences and demonstrate the potential of this array for fingerprinting several economically important members of this genus. In order to generate the Salvia subtracted diversity array (SDA) a suppression subtractive hybridization (SSH) was performed between a pool of Salvia species and a pool of angiosperms and non-angiosperms to selectively isolate Salvia-specific sequences. A total of 285-subtracted genomic DNA (gDNA) fragments were amplified and arrayed. DNA fingerprints were obtained for fifteen Salvia genotypes including three that were not part of the original subtraction pool. Hierarchical cluster analysis indicated that the Salvia-specific SDA was capable of differentiating S. officinalis and S. miltiorrhiza from their closely related species and was also able to reveal genetic relationships consistent with geographical origins. In addition, this approach was capable of isolating highly polymorphic sequences from chloroplast and nuclear DNA without preliminary sequence information. Therefore, SDA is a powerful technique for fingerprinting non-model plants and for identifying new polymorphic loci that may be developed as potential molecular markers.
鼠尾草是唇形科的一个重要属，约有 1000 种。该属分布于全球，被广泛用于观赏、烹饪和药用。我们报告了第一个富含多态性和发散性 DNA 序列的鼠尾草属指纹图谱阵列的构建，并展示了该阵列在鉴定该属几个经济重要成员方面的潜力。为了生成鼠尾草消减多样性阵列（SDA），我们在一组鼠尾草物种和一组被子植物和非被子植物之间进行了抑制性消减杂交（SSH），以选择性地分离出鼠尾草特异性序列。共扩增和排列了 285 个消减基因组 DNA（gDNA）片段。对包括三个未参与原始消减池的 15 个鼠尾草基因型进行了 DNA 指纹分析。层次聚类分析表明，鼠尾草特异性 SDA 能够区分丹参和紫丹参与其亲缘关系较近的物种，并且还能够揭示与地理起源一致的遗传关系。此外，这种方法能够从叶绿体和核 DNA 中分离出高度多态性的序列，而无需事先的序列信息。因此，SDA 是一种用于非模式植物指纹分析和鉴定新的多态性基因座的强大技术，这些基因座可能被开发为潜在的分子标记。
