Graphic Abstract

Establishment of quantitative PCR methods for the quantification of geosmin-producing potential and *Anabaena* sp. in freshwater systems

摘要

Geosmin has often been associated with off-flavor problems in drinking water with Anabaena sp. as the major producer. Rapid on-site detection of geosmin-producers as well as geosmin is important for a timely management response to potential off-flavor events. In this study, quantitative polymerase chain reaction (qPCR) methods were developed to detect the levels of Anabaena sp. and geosmin, respectively, by designing two PCR primer sets to quantify the rpoC₁ gene (ARG) and geosmin synthase one (GSG) in Anabaena sp. in freshwater systems. The ARG density determined by qPCR assay is highly related to microscopic cell count (r² = 0.726, p < 0.001), and the limit of detection (LOD) and limit of quantification (LOQ) of the qPCR method were 0.02 pg and 0.2 pg of DNA, respectively. At the same time, the relationship between geosmin concentrations measured by gas chromatography–mass spectrometry (GC–MS) and GSG copies was also established (r² = 0.742, p < 0.001) with similar LOD and LOQ values. Using the two qPCR protocols, we succeeded in measuring different levels of ARG and GSG copies in different freshwater systems with high incidence environmental substrata and diverse ecological conditions, showing that the methods developed could be applied for environmental monitoring. Moreover, comparing to the microscopic count and GC–MS analytical methods, the qPCR methods can reduce the time-to-results from several days to a few hours and require considerably less traditional algal identification and taxonomic expertise.

出版物
In Water Research.

Highlights

  • We designed primers targeting for Anabaena rpoC₁ gene and geosmin synthase gene.
  • We constructed two qPCR assays aim for quantifying general Anabaena population and geosmin-produce potential.
  • Both qPCR were verified by simulated culture bloom and field samples, hence are applicable.
  • Compared to traditional methods, the two qPCR assays provide fast, simple and on-site approach.
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苏命
副研究员

我的研究方向:水源地水质安全保障,主要针对水源地由于藻类爆发引起的水体嗅味问题开展研究。

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