Quantification of an arbuscular mycorrhizal fungus, Glomus mosseae, within plant roots by competitive polymerase chain reaction
Quantification of an arbuscular mycorrhizal fungus, Glomus mosseae, within plant roots by competitive polymerase chain reaction
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通过竞争性聚合酶链反应对植物根部的丛枝菌根真菌 Glomus mosseae 进行定量
DOI：
10.1017/s0953756297004218
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发表时间：
1997
期刊：
影响因子：
2.5
通讯作者：
J. Young
中科院分区：
文献类型：
--
作者：
S. Edwards;A. Fitter;J. Young
建立了一种基于竞争性聚合酶链式反应(PCR)的方法来定量测定植物根系中的丛枝菌根真菌(AM)--苔藓球菌。利用设计的甘蓝假单胞菌特异引物，在低严格的退火条件下扩增假单胞菌DNA，构建了异源内标。联合扩增大蒜根部提取物和内标DNA，可准确定量目标DNA。通过竞争性聚合酶链式反应和传统的定量方法显微镜，在一项比较研究中监测了大蒜根部的定植情况。这两种方法提供了在6周内从三个不同接种量水平获得的近乎平行的苔藓棉定殖率数据。结果表明，竞争性聚合酶链式反应是一种灵敏、准确的定量方法。竞争性聚合酶链式反应与显微镜相比的主要优势是它可以对特定的AM真菌进行定量。
An assay based on the competitive polymerase chain reaction (PCR) was developed to quantify Glomus mosseae , an arbuscular mycorrhizal (AM) fungus, within plant roots. Using previously designed G. mosseae specific primers, a heterologous internal standard was constructed by amplifying Pseudomonas DNA under low stringency annealing conditions. Co-amplification of G. mosseae and internal standard DNA within leek root extracts provided accurate quantification of target DNA. Colonization of leek roots by G. mosseae was monitored in a comparative study by competitive PCR and microscopy, a conventional method of quantification. These two methods gave closely parallel data for G. mosseae colonization from three different inoculum levels over a 6 week period. Results indicate that competitive PCR is a sensitive and accurate method of quantification. The major advantage of competitive PCR over microscopy is that it can quantify specific AM fungi.