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产菌氨酸类似物(MAAs)的铜绿微囊藻在伊利湖中的研究:定量 PCR 检测方法的建立及其生态学研究。

Mycosporine-like amino acids (MAAs)-producing Microcystis in Lake Erie: Development of a qPCR assay and insight into its ecology.

机构信息

Division of Environmental Health Sciences, College of Public Health, The Ohio State University, Columbus, OH, USA.

Aquatic Ecology Laboratory, Department of Evolution, Ecology and Organismal Biology, The Ohio State University, Columbus, OH, USA.

出版信息

Harmful Algae. 2018 Jul;77:1-10. doi: 10.1016/j.hal.2018.05.010. Epub 2018 Jun 1.

Abstract

Mycosporine-like amino acids (MAAs) are UV-absorbing metabolites found in cyanobacteria. While their protective role from UV in Microcystis has been studied in a laboratory setting, a full understanding of the ecology of MAA-producing versus non-MAA-producing Microcystis in natural environments is lacking. This study presents a new tool for quantifying MAA-producing Microcystis and applies it to obtain insight into the dynamics of MAA-producing and non-MAA-producing Microcystis in Lake Erie. This study first developed a sensitive, specific TaqMan real-time PCR assay that targets MAA synthetase gene C (mysC) of Microcystis (quantitative range: 1.7 × 10 to 1.7 × 10 copies/assay). Using this assay, Microcystis was quantified with a MAA-producing genotype (mysC) in water samples (n = 96) collected during March-November 2013 from 21 Lake Erie sites (undetectable - 8.4 × 10 copies/ml). The mysC genotype comprised 0.3-37.8% of the Microcystis population in Lake Erie during the study period. The proportion of the mysC genotype during high solar UV irradiation periods (mean = 18.8%) was significantly higher than that during lower UV periods (mean = 9.7%). Among the MAAs, shinorine (major) and porphyra (minor) were detected with HPLC-PDA-MS/MS from the Microcystis isolates and water samples. However, no significant difference in the MAA concentrations existed between higher and lower solar UV periods when the MAA concentrations were normalized with Microcystis mysC abundance. Collectively, this study's findings suggest that the MAA-producing Microcystis are present in Lake Erie, and they may be ecologically advantageous under high UV conditions, but not to the point that they exclusively predominate over the non-MAA-producers.

摘要

我的克肖酸类似氨基酸(MAAs)是在蓝藻中发现的紫外线吸收代谢物。虽然已经在实验室环境中研究了微囊藻中 MAAs 对紫外线的保护作用,但对自然环境中产生 MAAs 的微囊藻与不产生 MAAs 的微囊藻的生态学了解还不够充分。本研究提出了一种新的工具来量化产生 MAAs 的微囊藻,并应用该工具深入了解安大略湖产生 MAAs 的微囊藻和不产生 MAAs 的微囊藻的动态。

本研究首先开发了一种敏感、特异的 TaqMan 实时 PCR 检测方法,该方法靶向微囊藻的 MAAs 合成酶基因 C(mycS)(定量范围:1.7×10 至 1.7×10 拷贝/分析)。使用该检测方法,在 2013 年 3 月至 11 月期间从安大略湖 21 个地点采集的水样(不可检测-8.4×10 拷贝/ml)中定量检测到具有 MAAs 产生基因型(mycS)的微囊藻(n=96)。在研究期间,mycS 基因型占安大略湖微囊藻种群的 0.3-37.8%。在高太阳紫外线辐射期间(平均值=18.8%),mycS 基因型的比例明显高于低紫外线辐射期间(平均值=9.7%)。

在 HPLC-PDA-MS/MS 从微囊藻分离物和水样中检测到 shinorine(主要)和 porphyra(次要)作为 MAAs。然而,当将 MAAs 浓度与微囊藻 mycS 丰度归一化时,高太阳紫外线和低太阳紫外线期间的 MAAs 浓度没有显著差异。

综上所述,本研究的结果表明,产生 MAAs 的微囊藻存在于安大略湖中,它们在高 UV 条件下可能具有生态优势,但并没有使其完全占据不产生 MAAs 的微囊藻。

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