Identification of consumed stone flounder, Kareius bicoloratus (Basilewsky), from the stomach contents of sand shrimp, Crangon affinis (De Haan) using mitochondrial DNA analysis
Identification of consumed stone flounder, Kareius bicoloratus (Basilewsky), from the stomach contents of sand shrimp, Crangon affinis (De Haan) using mitochondrial DNA analysis
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DOI：
10.1016/s0022-0981(97)00039-7
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发表时间：
1997-10-01
影响因子：
2
通讯作者：
Kobayashi, T
中科院分区：
文献类型：
--
作者：
Asahida, T;Yamashita, Y;Kobayashi, T
建立了一种从沙虾Crangon affinis（De哈安）胃内容物中鉴别食用石鲽Kareius bicoloratus（Basilewsky）仔、稚鱼的方法。我们成功地分离石鲽DNA的胃内容物的沙虾，使用细胞裂解缓冲液含有8 M尿素。即使在捕食结束后5小时分离胃内容物，也可从胃内容物中获得用于PCR分析的DNA。然而，总DNA产量下降，捕食后的时间进行。利用鱼类通用引物PCR和线粒体DNA限制性内切酶分析，从沙虾胃内容物中鉴别出石鲽。石鲽mtDNA的成功扩增是可能的，直到4小时后，捕食终止，但没有扩增是可能的5小时后，捕食终止。此外，还克隆了石鲽mtDNA的部分D-loop区，并进行了序列测定，设计了石鲽种特异性PCR引物。PCR产物没有获得从其他可能的猎物的沙虾的总DNA：11种鱼类，2糠虾种，一个端足类物种，和2多毛类物种。本文讨论了PCR结合限制性酶切分析方法和物种特异性PCR方法鉴定捕食者肠道中被消耗的仔幼鱼物种的优缺点。(C)1997年Elsevier Science B.V.
We have developed a method for the identification of consumed stone flounder, Kareius bicoloratus (Basilewsky) larvae and juveniles from the stomach contents of sand shrimp, Crangon affinis (De Haan). We succeeded in isolating stone flounder DNA from the stomach contents of sand shrimp, using a cell lysis buffer containing 8 M urea. DNA for PCR analysis was obtained from stomach contents even when it was isolated 5 hours after the termination of predation. However, the total DNA yield decreased as the time after predation progressed. Stone flounder was distinguished from other prey in the stomach contents of sand shrimp using PCR with universal primers for fish and restriction analysis of mtDNA. Successful amplification of stone flounder mtDNA was possible up until 4 h after the termination of predation, but no amplification was possible 5 h after the termination of predation. In addition, a part of the D-loop region of stone flounder mtDNA was cloned, sequenced, and used to design species-specific PCR primers that allow amplification for stone flounder. PCR products were not obtained from the total DNA of other possible prey of the sand shrimp: 11 fish species; 2 mysid species; one amphipod species; and 2 polychaete species. In this paper, we discuss the advantages and disadvantages of using the PCR method coupled with restriction analysis, and the species-specific PCR method to identify consumed larval and juvenile fish species in the guts of predators. (C) 1997 Elsevier Science B.V.