Lozano-Duque Yesid, Richlen Mindy L, Smith Tyler B, Anderson Donald M, Erdner Deana L
Marine Science Institute - University of Texas at Austin, Port Aransas, USA.
Biology Department, Woods Hole Oceanographic Institution, Woods Hole, Massachusetts, USA.
J Appl Phycol. 2018 Dec;30(6):3529-3540. Epub 2018 May 7.
The genus is a recognized group of marine epiphytic-benthic dinoflagellates that produce the toxins that cause ciguatera fish poisoning (CFP). To date, thirteen species and six ribotypes of have been identified, and multiple species commonly co-occur within a single site or epiphyte community. Toxicity can vary by species, thus it is important to be able to differentiate among species for research and monitoring purposes. species have very similar morphological characteristics and are difficult or impossible to distinguish using light microscopy. DNA sequencing has been an important tool in the definition of species, but it can be time-consuming and relatively expensive. To provide an alternative approach, a PCR-RFLP protocol was developed for efficient, rapid, and cost-effective identification of strains isolated from the Gulf of Mexico and Caribbean Sea, where CFP cases and spp. have been reported. The assay targets the D1-D2 hypervariable regions of the large subunit ribosomal RNA gene and uses a single restriction enzyme, BsrI. This method produces distinct RFLP banding patterns for the six species of reported from the Gulf of Mexico and Caribbean Sea, and also distinguishes them from four Pacific endemic species. This method was successfully used to type 465 clonal isolates of from the U.S. Virgin Islands and Akumal Beach - Mexico This BsrI PCR-RFLP method expands the tools available to researchers and managers engaged in monitoring activities and ecological studies.
该属是一类公认的海洋附生底栖甲藻，它们产生的毒素会导致雪卡鱼中毒（CFP）。迄今为止，已鉴定出该属的13个物种和6种核糖型，并且多个物种通常在单个地点或附生植物群落中共存。毒性可能因物种而异，因此出于研究和监测目的，能够区分不同物种非常重要。该属物种具有非常相似的形态特征，使用光学显微镜很难或无法区分。DNA测序一直是定义该属物种的重要工具，但它可能耗时且相对昂贵。为了提供一种替代方法，开发了一种PCR-RFLP方案，用于高效、快速且经济高效地鉴定从墨西哥湾和加勒比海分离的该属菌株，在这些地区已报告了雪卡鱼中毒病例和该属物种。该检测针对大亚基核糖体RNA基因的D1-D2高变区，并使用单一限制酶BsrI。该方法为从墨西哥湾和加勒比海报告的6种该属物种产生了独特的RFLP条带模式，还将它们与4种太平洋特有物种区分开来。该方法已成功用于对来自美属维尔京群岛和墨西哥阿库马尔海滩的465株该属克隆分离株进行分型。这种BsrI PCR-RFLP方法扩展了从事监测活动和生态研究的研究人员和管理人员可用的工具。
