使用两种方法(生物传感器和实时 PCR)利用历史样本绘制选定的新出现的海洋产毒生物图谱:分辨率比较

Gerado Mengs, Rowena F. Stern, J. Clarke, Matthew Faith, Linda K. Medlin
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引用次数: 0

摘要

浮游生物连续记录仪(CPR)调查范围广、时间长,是绘制浮游生物分布变化图和了解有害藻类生态学的宝贵资源。使用福尔马林保存 DNA 的方法会使其降解,因此很难将其用作存档海洋样本的分子工具。从伊比利亚半岛沿线的巡航轨迹采集的 CPR 样品中,分别提取了采集后立即、7 个月后和保存 9 年后的 DNA。将九年时间点的 PCR 反应与电化学生物传感器中的探针杂交,并与两个较早时间点的 RT-PCR 结果进行比较。结果表明,成功地鉴定出了 Pseudo-nitzschia spp.、Prorocentrum lima、Alexandrium minutum、Alexandrium ostenfeldii、Gambierdiscus spp.和 Coolia spp.。生物传感器的分析结果优于 RT-PCR,使我们能够记录某些热带有毒甲藻,即 Gambierdiscus 和 Coolia,它们分别产生人类雪卡毒素和 Coolia 毒素。这些非本地藻类毒素会积累起来,渗透到食物网中,对人类的粮食安全产生负面影响。这证明随着伊比利亚半岛近海水域的气候变化,微藻会向北移动。这项研究强调了生物传感器是近海监测有害藻类繁殖的一种经济有效的工具,并推动了分子技术在有害藻类记录有限的长期氯化石蜡数据集上的应用。福尔马林保存的 CPR 样本中的 DNA 会降解,因此使用短小的多探针生物传感器可以利用现代方法增加浮游生物的历史记录,同时还能捕获地表水中不常出现的底栖生物分类群。基于探针的生物传感器技术的集成为探索浮游生物响应环境变化的动态提供了一个前景广阔的途径。
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Mapping Selected Emergent Marine Toxin-Producing Organisms Using Historical Samples with Two Methods (Biosensors and Real-Time PCR): A Comparison of Resolution
The Continuous Plankton Recorder (CPR) survey is a valuable resource for mapping changes in plankton distribution and understanding harmful algal ecology because of its breadth and longevity. Preservation methods with formalin degrade DNA, making it difficult to use as a molecular tool for archived marine samples. DNA was extracted from CPR samples immediately after collection, seven months later and after nine years of storage from a cruise track along the Iberian Peninsula. PCR reactions performed from the nine-year timepoint were hybridized to probes in an electrochemical biosensor and compared to results obtained from RT-PCR performed at two earlier time points. The successful identification of Pseudo-nitzschia spp., Prorocentrum lima, Alexandrium minutum, Alexandrium ostenfeldii, Gambierdiscus spp. and Coolia spp. was documented. The biosensor analysis outperformed RT-PCR, allowing us to document certain tropical toxic dinoflagellates, viz., Gambierdiscus and Coolia, that produce human ciguatoxins and Coolia toxins, respectively. These non-native algal toxins can accumulate, pervade the food web and negatively impact human food security. This supports the northerly movement of microalgae with climate change in offshore Iberian peninsular waters. This study highlights biosensors as a cost-effective tool for the offshore monitoring of HAB species and advances molecular technologies for long-term CPR datasets that have limited records of harmful algae. DNA from formalin-preserved CPR samples is degraded, so the use of a short, multiprobe biosensor can augment historical plankton records with contemporary methods that also capture infrequently occurring benthic taxa carried in surface waters. The integration of probe-based biosensor technologies offers a promising avenue for exploring plankton dynamics in response to environmental changes.
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