The United Graduate School of Agriculture Science, Gifu University, Gifu 501-1193, Japan.
Microbiol Res. 2010 Oct 20;165(8):695-705. doi: 10.1016/j.micres.2009.11.010. Epub 2010 Jan 29.
Pythium intermedium is known to play an important role in the carbon cycling of cool-temperate forest soils. In this study, a fast, precise and effective real-time PCR technique for estimating the population densities of P. intermedium from soils was developed using species-specific primers. Specificity was confirmed both with conventional PCR and real-time PCR. The detection limit (sensitivity) was determined and amplification standard curves were generated using SYBR Green II fluorescent dye. A rapid and accurate assay for quantification of P. intermedium in Takayama forest soils of Japan was developed using a combination of a new DNA extraction method and PCR primers were developed for real-time PCR. And the distribution of P. intermedium in forest soil was investigated with both soil plating method and the developed real-time PCR technique. This new technique will be a useful tool and can be applied to practical use for studying the role of Pythium species in forest and agricultural ecosystems.
中间茎点霉在冷温带森林土壤的碳循环中起着重要作用。本研究采用物种特异性引物，开发了一种快速、精确、有效的实时 PCR 技术，用于估计土壤中中间茎点霉的种群密度。通过常规 PCR 和实时 PCR 均证实了特异性。通过 SYBR Green II 荧光染料确定了检测限（灵敏度），并生成了扩增标准曲线。采用一种新的 DNA 提取方法和用于实时 PCR 的引物，建立了一种快速准确的定量日本高山森林土壤中间茎点霉的方法。采用土壤平板法和开发的实时 PCR 技术研究了中间茎点霉在森林土壤中的分布。该新技术将成为一种有用的工具，并可应用于研究茎点霉属在森林和农业生态系统中的作用的实际应用。
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