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首页> 外文期刊>Aquatic Microbial Ecology >Detection of the bloom-forming cold-water dinoflagellate Biecheleria baltica in the Baltic Sea using LSU rRNA probes
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Detection of the bloom-forming cold-water dinoflagellate Biecheleria baltica in the Baltic Sea using LSU rRNA probes

机译:使用LSU rRNA探针检测波罗的海中形成水华的水生鞭毛藻(Biecheleria baltica)

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摘要

ABSTRACT: Cold-water dinoflagellates frequently dominate the spring phytoplankton community of the northern Baltic Sea and contribute substantially to the spring primary production. These dinoflagellate communities are largely composed of 3 different species (Biecheleria baltica, Scrippsiella hangoei, Gymnodinium corollarium) that cannot be unambiguously separated by conventional light microscopy. In this study, a fluorescence in situ hybridization (FISH) assay was developed for the detection of B. baltica in field samples. The probe-based method was tested and optimized in vitro, and potential effects of the B. baltica life-cycle transitions were evaluated by examining the labeling quality of the probes on different life-cycle stages before the assay was applied in a field study. The BbRNA4 probe had the highest specificity and was chosen for downstream applications. The life-cycle experiment showed significant differences in labeling efficiency between cultured cells from different growth phases, particularly for the nutrient-limited treatment compared to the control. Tests with spiked field samples revealed that cell recovery in the FISH assay was low (30%), resulting in a considerable underestimation of in situ abundances. However, a strong relationship between FISH and Utermöhl counts of field samples suggests that trends of the bloom dynamics can be followed reliably by this method. By applying the FISH detection method in a field survey, we found that the dynamics of the B. baltica bloom follows that established earlier for the Scrippsiella/Biecheleria/ Gymnodinium complex, with 2 peaks throughout the season. When corrected for processing losses, FISH-based abundance estimates suggest that B. baltica comprised the major fraction of the total dinoflagellate complex during the spring bloom at the southwest coast of Finland.
机译:摘要:冷水鞭毛虫经常在波罗的海北部的春季浮游植物群落中占主导地位,并为春季的初级生产做出重要贡献。这些鞭毛鞭毛虫群落主要由3种不同的物种组成( Balcheleria baltica , Scrippsiella hangoei Gymnodinium花冠),这些物种无法通过常规光学显微镜明确区分。 。在这项研究中,开发了一种荧光原位杂交(FISH)测定法来检测 B。现场样品中的波罗的海。测试并优化了基于探针的方法体外以及 B的潜在作用。在将分析应用于现场研究之前,通过检查探针在不同生命周期阶段的标记质量来评估baltica的生命周期过渡。 BbRNA4探针具有最高的特异性,并被选择用于下游应用。生命周期实验显示,来自不同生长期的培养细胞之间的标记效率存在显着差异,尤其是与对照相比,营养受限的处理更是如此。对加标的野外样品进行的测试表明,FISH分析中的细胞回收率很低(30%),从而大大低估了原位丰度。但是,FISH和田间样品的Utermöhl计数之间有很强的关系,表明该方法可以可靠地跟踪水华动态。通过在现场调查中应用FISH检测方法,我们发现 B的动力学。波罗的海盛开的花朵早于S草/白屈菌/裸子草复合体,在整个季节都有2个高峰。如果对加工损失进行校正,则基于FISH的丰度估计值表明B。在芬兰西南海岸的春季开花期间,波罗的海占总鞭毛虫复合物的大部分。

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