大黄鱼微卫星标记引物在石首鱼科几个近缘种中的通用性研究

Cross-species amplification of microsatellite markers from Pseudosciaena crocea in Sciaenidae

本研究分析了8对大黄鱼(Pseudosciaena crocea)微卫星标记引物在石首鱼科5个亚科8属10个种中的通用性。结果表明,大黄鱼微卫星标记引物在黄鱼亚科鱼类中有较高的通用性,其中位点PC1C4,PC4H12,PC5E11,PC8F5,PC10F10,PC10G6的引物可在小黄鱼(P.polyactis)和棘头梅童鱼(Collichthys lucidus)中获得清晰扩增条带,位点PC7H4引物也可在小黄鱼中扩增,上述可扩增位点在两个种中均得到多个等位基因,表现出较高的多态性;大黄鱼微卫星标记引物在白姑鱼亚科、叫姑鱼亚科、牙亚科及石首鱼亚科鱼类中也有较高的通用性,其中在银牙(Otolithes argenteus)中得到6个可扩增位点(PC1C4,PC4H12,PC7A2,PC7H4,PC10F10,PC10G6),在鮸状黄姑鱼(Nibea miichthioides)、丁氏(Wak tingi)和眼斑拟石首鱼(Sciaenops ocellatus)中各得到5个可扩增位点,在浅色黄姑鱼(N.Chui)和尖头黄鳍牙(Chrysochir aureus)中各得到4个可扩增位点,在皮氏叫姑鱼(Johnius belengerii)中得到3个可扩增位点(PC7H4,PC10F10,PC10G6)。位点PC10F10和PC10G6引物在10个种中都可得到清晰的扩增产物,可作为10种石首鱼的通用标记。引物PC5E11和PC8F5只在黄鱼亚科中得到扩增产物,为黄鱼亚科特异性标记。

The objectives of this study were to assess the ability of 8 pairs of microsatellite primers developed from Pseudosciaena crocea to amplify microsatellites in ten species of Sciaenidae fishes which were collected from coast of Zhejiang Province and Fujian Province. It was found that the rate of successful amplification in species belonging to the same genus or the same subfamily as the target species was high. In the subfamily of Pseudosciaeninae, 7 loci （ PC1C4, PC4H12, PC5E11, PCTH4, PC8F5, PC10F10, PC10G6 ） were amplified successfully in P. polyactis, and 6 loci were amplified successfully in Collichthys lucidus exoept PC4H12 and PCTA2. These loci which could be amplified revealed polymorphism in Pseudosciaeninae. Primers of these loci were also useful in species ofArgyrosominae, Johniinae, Otolithinae and Sciaeninae. 6 loci （PC1C4, PC4H12, PC7A2, PCTH4, PC10F10, PC10G6） of 8 loci were amplified successfully in Otolithes argenteus. Loci PC1C4, PC4H12, PCTH4, PC10F10, PC10G6 could be amplified successfully in Nibea miichthioides and Sciaenops ocellatus, while PC1C4, PC7A2, PCTH4, PC10F10, PC10G6 in Wak tingi. Loci PC1C4, PC7A2, PC10F10, PC10G6 and loci PC1C4, PCTH4, PC10F10, PC10G6 have been amplified in N. chui and Chrysochir aureus, respectively. Meanwhile, only 3 loci （PCTH4, PC10F10, PC10G6）were amplified suecessfully in Johnias belengerii. Our cross-species amplification data indicated that allelic diversity was very low in other species compared with P crocea. For example, locus PCIOFIO has high allelic diversity in P. crocea but it has only one allele in S. ocellatus. Loci PC10F10 and PC10G6 were extremely conservative, they could be amplified in all species used in the test. The result suggested that these two loci had potential value in inter-specific molecular comparisons and phylogeny reconstruction among different Sciaenidae fishes. Loci PC5E11 and PC8F5 were only amplified in subfamily Pseudosciaeninae. Maybe they were specific loci in Pseudosciaeninae, which indicated we could use these loci to identify species of Pseudosciaeninae from other Sciaenidae fishes. In additional, we found that there were differences between closely related Sciaenidae fishes in average allelic length and the number of alleles which had been scored, thus the microsatellite＇ s footprints of each taxonomic species might be used to discriminate closely related species, but it need more data to confirm the inferrer. [Journal of Fishery Sciences of China, 2008, 15 （2） ： 237-243]