Jérôme Marc, Macé Sabrina, Dousset Xavier, Pot Bruno, Joffraud Jean-Jacques
Ifremer, Laboratoire Ecosystèmes Microbiens et Molécules Marines pour les Biotechnologies (EM(3)B), BP21105, 44311 Nantes, France.
University of Liège, Food Science Department, Fundamental and Applied Research for Animal and Health, Food Microbiology, Sart-Tilman B43b, B-4000 Liège, Belgium.
Int J Food Microbiol. 2016 Jan 18;217:101-9. doi: 10.1016/j.ijfoodmicro.2015.10.018. Epub 2015 Oct 19.
An accurate amplified fragment length polymorphism (AFLP) method, including three primer sets for the selective amplification step, was developed to display the phylogenetic position of Photobacterium isolates collected from salmon products. This method was efficient for discriminating the three species Photobacterium phosphoreum, Photobacterium iliopiscarium and Photobacterium kishitanii, until now indistinctly gathered in the P. phosphoreum species group known to be strongly responsible for seafood spoilage. The AFLP fingerprints enabled the isolates to be separated into two main clusters that, according to the type strains, were assigned to the two species P. phosphoreum and P. iliopiscarium. P. kishitanii was not found in the collection. The accuracy of the method was validated by using gyrB-gene sequencing and luxA-gene PCR amplification, which confirmed the species delineation. Most of the isolates of each species were clonally distinct and even those that were isolated from the same source showed some diversity. Moreover, this AFLP method may be an excellent tool for genotyping isolates in bacterial communities and for clarifying our knowledge of the role of the different members of the Photobacterium species group in seafood spoilage.
开发了一种准确的扩增片段长度多态性（AFLP）方法，该方法在选择性扩增步骤中包括三组引物，用于显示从鲑鱼产品中分离出的发光杆菌菌株的系统发育位置。这种方法对于区分磷光发光杆菌、鱼发光杆菌和岸田发光杆菌这三个物种非常有效，到目前为止，它们一直被模糊地归为磷光发光杆菌物种组，而该物种组被认为是海鲜腐败的主要原因。AFLP指纹图谱能够将分离菌株分为两个主要聚类，根据模式菌株，这两个聚类分别被归为磷光发光杆菌和鱼发光杆菌这两个物种。在该样本中未发现岸田发光杆菌。通过使用gyrB基因测序和luxA基因PCR扩增验证了该方法的准确性，这证实了物种的划分。每个物种的大多数分离菌株在克隆上是不同的，即使是从同一来源分离出的菌株也表现出一定的多样性。此外，这种AFLP方法可能是对细菌群落中的分离菌株进行基因分型以及明确我们对发光杆菌物种组不同成员在海鲜腐败中所起作用的认识的一种优秀工具。
