Jones Matthew W, O'Reilly Patrick T, McPherson Arran A, McParland Tara L, Armstrong Dawn E, Cox Andrea J, Spence Koren R, Kenchington Ellen L, Taggart Chris T, Bentzen Paul
Marine Gene Probe Laboratory, Dalhousie University, Halifax, NS B3H 4J1, Canada.
Genome. 2003 Feb;46(1):59-69. doi: 10.1139/g02-105.
Effective management of exploited species demands contemporary knowledge of population structure and mating patterns. Genetic markers can prove useful in providing this knowledge. Despite its commercial importance, genetic markers for American lobster (Homarus americanus) are limited. We developed 12 tetra- and 1 trinucleotide microsatellite loci for American lobster that exhibit little stuttering after PCR amplification. Gene diversity of these loci ranged from 0.516 to 0.929. A four-locus multiplex permits rapid genotyping of progeny in parentage experiments with a paternity exclusion probability over the four loci of 97.8%. We examined the loci for conformity to Hardy-Weinberg expectations (HWE) and linkage using individuals from one location and found that four loci deviated from HWE. We also tested inheritance and pairwise linkage using 48 embryos from each of two females. With the exception of two loci that were derived from the same clone and separated by 72 bp, no evidence of linkage was found. We, for the first time, demonstrate the occurrence of multiple paternity in American lobster. We also observed an apparent occurrence of dispermic androgenesis, possibly the first documentation of such an event within a species. Ten of the loci amplified in European lobster (Homarus gammarus), although two were monomorphic and one deviated significantly from HWE. We quantified mitochondrial DNA (mtDNA) sequence variation through the use of PCR amplification of two DNA fragments, followed by digestion with restriction enzymes; eight haplotypes were detected. One of the two fragments amplified in European lobster. Both sets of markers should prove useful for population discrimination purposes, and the microsatellites, in particular the four-locus multiplex, should prove highly amenable to rapidly addressing questions about mating patterns.
对受开发利用物种的有效管理需要掌握有关种群结构和交配模式的当代知识。遗传标记有助于提供此类知识。尽管美洲龙虾具有商业重要性，但其遗传标记却很有限。我们开发了12个四核苷酸和1个三核苷酸微卫星位点用于美洲龙虾，这些位点在PCR扩增后几乎没有拖尾现象。这些位点的基因多样性范围为0.516至0.929。一个四位点多重体系允许在亲权实验中对后代进行快速基因分型，四个位点的父权排除概率超过97.8%。我们使用来自一个地点的个体检查这些位点是否符合哈迪-温伯格平衡期望（HWE）和连锁关系，发现四个位点偏离了HWE。我们还使用来自两只雌性龙虾的各48个胚胎测试了遗传和成对连锁关系。除了两个来自同一克隆且相隔72 bp的位点外，未发现连锁证据。我们首次证明了美洲龙虾中存在多重父权现象。我们还观察到明显的双精卵孤雌生殖现象，这可能是该物种内此类事件的首次记录。其中10个位点在欧洲龙虾（欧洲螯龙虾）中也能扩增，不过有两个位点是单态的，还有一个位点显著偏离HWE。我们通过PCR扩增两个DNA片段，然后用限制性酶消化来量化线粒体DNA（mtDNA）序列变异；检测到了8种单倍型。其中一个片段在欧洲龙虾中也能扩增。这两组标记对于种群鉴别都应是有用的，特别是微卫星标记，尤其是四位点多重体系，对于快速解决有关交配模式的问题应是非常适用的。
