Saju J M, Németh Sz, Szűcs Réka, Sukumaran Rashmi, Lim Z, Wong L, Orbán L, Bercsényi M
National University of Singapore Reproductive Genomics Group, Strategic Research Program, Temasek Life Sciences Laboratory, 1 Research Link Singapore 117604 Singapore.
University of Pannonia Department of Animal Sciences and Breeding, Georgikon Faculty Deák Ferenc u. 16 H-8230 Keszthely Hungary.
Acta Biol Hung. 2014 Jun;65(2):132-43. doi: 10.1556/ABiol.65.2014.2.2.
The identification of three scorpionfish species, the black scorpionfish (Scorpaena porcus Linnaeus, 1758), the large-scaled scorpionfish (S. scrofa Linnaeus, 1758) and the small red scorpionfish (S. notata Rafinesque, 1810) is possible in adults by morphometry, but often problematic in juveniles due to their similar phenotypes. To develop a molecular species identification tool, first, we have analyzed the genetic similarity of the three species by a PCR-based 'blind method' that amplified bands from various locations of the genome. We found high levels of nucleotide similarity between S. porcus and S. scrofa, whereas S. notata showed a higher level of divergence from the other two species. Then, we have searched these patterns for differences between the genomes of Adriatic specimen of these three species and identified several species-specific products in two of them. For the third one a species-specific primer pair amplifying from the 16S ribosomal DNA was designed. One marker for each species was cloned, sequenced and converted into Sequence Characterized Amplified Region (SCAR) markers amplified by specific primer pairs. The SCAR markers amplified robust bands of limited variability from the target species, while no or only occasional weak products were obtained from the other two, proving that they can be used for molecular identification of these three species. These markers can help the conservation and future analysis of these three species as well as their possible selection programs for aquaculture purposes.
通过形态测量法在成体中识别三种鲉鱼物种，即黑鲉（Scorpaena porcus Linnaeus，1758年）、大鳞鲉（S. scrofa Linnaeus，1758年）和小红鲉（S. notata Rafinesque，1810年）是可行的，但在幼体中往往存在问题，因为它们的表型相似。为了开发一种分子物种识别工具，首先，我们通过一种基于PCR的“盲法”分析了这三种物种的遗传相似性，该方法从基因组的不同位置扩增条带。我们发现黑鲉和大鳞鲉之间的核苷酸相似性水平很高，而小红鲉与其他两个物种的差异水平更高。然后，我们在这三种物种的亚得里亚海标本的基因组之间寻找这些模式的差异，并在其中两种中鉴定出几种物种特异性产物。对于第三种，设计了一对从16S核糖体DNA扩增的物种特异性引物对。为每个物种克隆了一个标记，进行了测序，并将其转化为由特异性引物对扩增的序列特征扩增区域（SCAR）标记。SCAR标记从目标物种中扩增出变异性有限的稳健条带，而从其他两个物种中未获得或仅偶尔获得弱产物，这证明它们可用于这三种物种的分子鉴定。这些标记有助于这三种物种的保护和未来分析，以及它们可能用于水产养殖目的的选择计划。
