A universal molecular method for identifying underground plant parts to species
A universal molecular method for identifying underground plant parts to species
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DOI：
10.1046/j.1365-294x.2000.01034.x
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发表时间：
2000-10-01
影响因子：
4.9
通讯作者：
Jackson, RB
中科院分区：
文献类型：
--
作者：
Linder, CR;Moore, LA;Jackson, RB
作为确定德克萨斯州中部爱德华兹高原的根深和资源吸收大型项目的一部分，我们开发了一种基于 DNA 的技术，可以将所有植物的地下部分识别到属的水平，通常是物种的水平。通过将来自地下植物材料的 18S-26S 核核糖体 DNA 重复序列的内部转录间隔区 (ITS) 区域的 DNA 序列与某个地点植物的参考 ITS 区域数据库进行比较来实现鉴定。该方法适用于整个植物，因为可以使用一组通用引物对植物 ITS 区域进行 PCR 扩增。由于 ITS 区域进化相对较快，因此通常可以识别同属物种。在我们的研究中，所有的根都很容易被识别到属的水平；大多数同属物种仅通过 ITS 序列差异来识别，但有些物种需要结合 ITS 序列数据和对某个地点的物种进行地上调查。除了展示我们的技术的可行性和有效性之外，我们还将其与另一种用于识别地下植物部分的基于 DNA 的技术进行了比较。最后，我们还描述了一种 DNA 提取和纯化技术，该技术能够可靠地从根中提供足够数量的高质量 DNA，以便可以轻松完成 PCR。我们的技术应该能够识别任何系统中植物的地下部分，从而为地下植物群落的研究开辟新的可能性。
As part of a large project to determine rooting depth and resource uptake on the Edwards Plateau of central Texas, we developed a DNA-based technique that allows the below-ground parts of all plants to be identified to the level of genus and usually to species. Identification is achieved by comparing DNA sequences of the internal transcribed spacer (ITS) region of the 18S-26S nuclear ribosomal DNA repeat, derived from below-ground plant material, with a reference ITS region database for plants at a site. The method works throughout plants because the plant ITS region can be PCR amplified using a set of universal primers. Congeneric species can usually be identified because the ITS region evolves relatively rapidly. In our study, all roots were easily identified to the level of genus; most congeneric species were identified solely by ITS sequence differences but some required a combination of ITS sequence data and above-ground surveys of species at a site. In addition to showing the feasibility and efficacy of our technique, we compare it with another DNA-based technique used to identify belowground plant parts. Finally, we also describe a DNA extraction and purification technique that reliably provides high-quality DNA of sufficient quantity from roots so that PCR can be readily accomplished. Our technique should allow the below-ground parts of plants in any system to be identified and thereby open new possibilities for the study of below-ground plant communities.