PCR primers that amplify fungal rRNA genes from environmental samples
PCR primers that amplify fungal rRNA genes from environmental samples
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DOI：
10.1128/aem.66.10.4356-4360.2000
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发表时间：
2000-10-01
影响因子：
4.4
通讯作者：
Hartin, RJ
中科院分区：
文献类型：
--
作者：
Borneman, J;Hartin, RJ
设计了两对PCR引物，用于扩增子囊菌门（Ascomycota）、担子菌门（Basidiomycota）、壶菌门（Chyhridomycota）和接合菌门（Zygomycota）4个主要真菌门的rRNA基因（rDNA）。为了测试引物从环境样品中扩增真菌rDNA的能力，构建并分析了来自两个鳄梨格罗夫土壤的克隆文库。这两种土壤对肉桂疫霉根腐病的抑制能力不同。它还揭示了一个显着不同的描述土壤真菌群落比由基于培养的分析，确认的价值，基于rDNA的方法来识别可能不容易在琼脂培养基上生长的生物体。通过鉴定与鳄梨叶相关的真菌获得了引物有用性的其他证据。在土壤和叶片分析中，没有发现非真菌rDNA序列，说明这些PCR引物的选择性。这项工作证明了两个新开发的PCR引物组从土壤和植物组织中扩增真菌rDNA的能力，从而提供了独特的工具来研究这个庞大的，大多数未描述的生物群落。
Two PCR primer pairs were designed to amplify rRNA genes (rDNA) from all four major phyla of fungi: Ascomycota, Basidiomycota, Chyhridomycota, and Zygomycota, PCRs performed with these primers showed that both pairs amplify DNA from organisms representing the major taxonomic groups of fungi but not from nonfungal sources. To test the ability of the primers to amplify fungal rDNA from environment samples, clone libraries from two avocado grove soils were constructed and analyzed. These soils possess different abilities to inhibit avocado root rot caused by Phythophthora cinnamomi, Analysis of the two rDNA clone libraries revealed differences in the two fungal communities. It also revealed a markedly different depiction of the soil fungal community than that generated by a culture-based analysis, confirming the value of rDNA-based approaches for identifying organisms that may not readily grow on agar media. Additional evidence of the usefulness of the primers was obtained by identifying fungi associated with avocado leaves. In both the soil and leaf analyses, no nonfungal rDNA sequences were identified, illustrating the selectivity of these PCR primers. This work demonstrates the ability of two newly developed PCR primer sets to amplify fungal rDNA from soil and plant tissue, thereby providing unique tools to examine this vast and mostly undescribed community of organisms.