Gachon Claire M M, Strittmatter Martina, Müller Dieter G, Kleinteich Julia, Küpper Frithjof C
Culture Collection of Algae and Protozoa, Scottish Association for Marine Science, Dunstaffnage Marine Laboratory, Oban, Argyll PA37 1QA, Scotland, United Kingdom.
Appl Environ Microbiol. 2009 Jan;75(2):322-8. doi: 10.1128/AEM.01885-08. Epub 2008 Nov 14.
In the marine environment, a growing body of evidence points to parasites as key players in the control of population dynamics and overall ecosystem structure. However, their prevalence and impact on marine macroalgal communities remain virtually unknown. Indeed, infectious diseases of seaweeds are largely underdocumented, partly because of the expertise required to diagnose them with a microscope. Over the last few years, however, real-time quantitative PCR (qPCR) has emerged as a rapid and reliable alternative to visual symptom scoring for monitoring pathogens. Thus, we present here a qPCR assay suitable for the detection and quantification of the intracellular oomycete pathogen Eurychasma dicksonii in its ectocarpalean and laminarialean brown algal hosts. qPCR and microscopic observations made of laboratory-controlled cultures revealed that clonal brown algal strains exhibit different levels of resistance against Eurychasma, ranging from high susceptibility to complete absence of symptoms. This observation strongly argues for the existence of a genetic determinism for disease resistance in brown algae, which would have broad implications for the dynamics and genetic structure of natural populations. We also used qPCR for the rapid detection of Eurychasma in filamentous brown algae collected in Northern Europe and South America and found that the assay is specific, robust, and widely applicable to field samples. Hence, this study opens the perspective of combining large-scale disease monitoring in the field with laboratory-controlled experiments on the genome model seaweed Ectocarpus siliculosus to improve our understanding of brown algal diseases.
在海洋环境中,越来越多的证据表明寄生虫是控制种群动态和整体生态系统结构的关键因素。然而,它们在海洋大型藻类群落中的流行情况及其影响实际上仍不为人知。事实上,海藻的传染病在很大程度上记录不足,部分原因是需要显微镜诊断的专业知识。然而,在过去几年中,实时定量PCR(qPCR)已成为一种快速可靠的替代方法,可用于监测病原体,替代视觉症状评分。因此,我们在此展示一种qPCR检测方法,适用于检测和定量细胞内卵菌病原体迪克逊藻(Eurychasma dicksonii)在其外果皮藻和海带目褐藻宿主中的情况。对实验室控制培养物进行的qPCR和显微镜观察表明,克隆褐藻菌株对迪克逊藻表现出不同程度的抗性,从高敏感性到完全无症状。这一观察结果有力地证明了褐藻中存在抗病性的遗传决定因素,这将对自然种群的动态和遗传结构产生广泛影响。我们还使用qPCR快速检测在北欧和南美采集的丝状褐藻中的迪克逊藻,发现该检测方法具有特异性、稳健性,并且广泛适用于野外样本。因此,本研究开启了将野外大规模疾病监测与对基因组模型海藻——硅藻(Ectocarpus siliculosus)进行实验室控制实验相结合的前景,以增进我们对褐藻疾病的理解。