珠江三种鲌亚科鱼类微卫星鉴别技术的建立

Establishment of microsatellite identification on there Culterinae fishes in Pearl River

应用微卫星标记技术鉴别珠江3种主要鲌亚科鱼类广东鲂（Megalobrama terminalis）、鳊（Parabamis pekinensis）、海南红鲌（Culter recurviceps）.从GenBank和文献资料中初选3种鲌亚科鱼类的微卫星标记,设计108对微卫星引物,用PCR扩增3种鱼的基因组DNA,并用聚丙烯酰胺凝胶电泳检测选出每种鱼的特异微卫星标记,共获得16个特异性标记.其中,微卫星标记F-524在广东鲂和海南红鲌中扩增的目的片段为309～320 bp,在鳊中无扩增条带;微卫星标记L-4在鳊和海南红鲌中扩增目的片段为160～201 bp,在广东鲂中无扩增条带;微卫星标记L-7在广东鲂和鳊中扩增的目的片段为160～190 bp,在海南红鲌中扩增的片段为123～143 bp.3个微卫星标记,单独使用1个特异性标记或几种特异性微卫星标记相结合,可快速从分子水平鉴别出广东鲂、鳊、海南红鲌,从而解决3种鲌亚科鱼类早期发育过程中形态比较相似,通过形态鉴定比较困难的问题.

Microsatellite markers were used to distinguish Megalobrama terminalis, Parabramis pekinensis and Culter recurviceps in Pearl River. Using genomic of M. terminalis, P. pekinensis and C. recurviceps as templates, PCR amplifications were performed by using 108 pairs of primers which were designed based on Cuherinae fishes microsatellite markers selected from GenBank and literature, and 16 special microsatellite makers were selected through PAGE glue. The result showed that, the maker F-524 could PCR an amplified DNA fragment between 309-320 bp in M. terminalis and C. recurviceps; the maker L-4 could amplify an amplified DNA fragment between 160-201 bp in C. recurviceps and P. pekinensis; the maker L-7 could amplify an amplified DNA fragment between 160-190 bp in M. terminalis and P. pekinensis, and it could also amplify an amplified DNA fragment between 123-143 bp in C. recurviceps. In early growth, the morphological of M. terminalis, P. pekinensis and C. recurviceps were almost the same, thus, we couldn＇t identify them, but when we used one specific marker alone or combined several specific markers together, we could quickly identify these three species on molecular level.