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一种使用本地水生物种定量生物可利用磷的qPCR方法。

A qPCR method to quantify bioavailable phosphorus using indigenous aquatic species.

作者信息

Yang Yanan, Yang Jianghua, Zhang Xiaowei

机构信息

State Key Laboratory of Pollution Control & Resource Reuse, School of the Environment, Nanjing University, 163 Xianlin Avenue, Nanjing, 210023 China.

出版信息

Environ Sci Eur. 2018;30(1):32. doi: 10.1186/s12302-018-0163-z. Epub 2018 Sep 4.

Abstract

BACKGROUND

Bioavailable phosphorus (BAP) represents the sum of phosphorus that is readily available for algae growth and is useful to indicate the severity of eutrophication in aquatic environments.

RESULTS

Here, a quantitative real-time PCR (qPCR)-based bioassay was developed to quantify BAP using the indigenous cyanobacterium species sp. of Lake Tai, a large and shallow eutrophic lake in the Yangtze Valley, China. Primers were designed to quantify the gene expression of alkaline phosphatase (/- and phosphate transporter () genes of . The specificity and efficiency of the primer sets were evaluated by gel electrophoresis and real-time PCR. The results showed that the primers developed here could successfully be used to measure BAP in the water. The linear range of BAP measurements by the gene after 2 h incubation was 0.125-2.00 mg/L. Then, the qPCR-based bioassay was applied to analyze water samples from Tai Lake, which had BAP levels in the range of 0.239-0.459 mg/L.

CONCLUSIONS

The qPCR-based bioassay represents a promising biomonitoring tool that can quantify phosphorus bioavailability in aquatic environments.

摘要

背景

生物可利用磷(BAP)代表易于被藻类利用的磷的总和,有助于指示水生环境富营养化的严重程度。

结果

在此,开发了一种基于定量实时PCR(qPCR)的生物测定法,利用中国长江流域大型浅水富营养化湖泊太湖的本土蓝藻物种来定量BAP。设计引物以定量碱性磷酸酶(/ - )和磷酸盐转运蛋白()基因的基因表达。通过凝胶电泳和实时PCR评估引物组的特异性和效率。结果表明,此处开发的引物可成功用于测量水中的BAP。孵育2小时后,通过基因测量BAP的线性范围为0.125 - 2.00mg/L。然后,基于qPCR的生物测定法用于分析太湖的水样,其BAP水平在0.239 - 0.459mg/L范围内。

结论

基于qPCR的生物测定法是一种很有前景的生物监测工具,可以量化水生环境中的磷生物可利用性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3556/6132795/460e59905b90/12302_2018_163_Fig1_HTML.jpg

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