Paz-García David A, Munguía-Vega Adrián, Plomozo-Lugo Tomas, Weaver Amy Hudson
Department of Biological Sciences, Louisiana State University, Baton Rouge, LA, USA.
Laboratorio de Necton y Ecología de arrecifes. Centro de Investigaciones Biológicas del Noroeste, Instituto Politécnico Nacional, 195, Col. Playa Palo de Santa Rita Sur, La Paz, Baja California Sur, Mexico.
Mol Biol Rep. 2017 Apr;44(2):251-256. doi: 10.1007/s11033-017-4105-4. Epub 2017 Apr 27.
We developed a set of hypervariable microsatellite markers for the Pacific red snapper (Lutjanus peru), an economically important marine fish for small-scale fisheries in the west coast of Mexico. We performed shotgun genome sequencing with the 454 XL titanium chemistry and used bioinformatic tools to search for perfect microsatellite loci. We selected 66 primer pairs that were synthesized and genotyped in an ABI PRISM 3730XL DNA sequencer in 32 individuals from the Gulf of California. We estimated levels of genetic diversity, deviations from linkage and Hardy-Weinberg equilibrium, estimated the frequency of null alleles and the probability of individual identity for the new markers. We reanalyzed 16 loci in 16 individuals to estimate genotyping error rates. Eighteen loci failed to amplify, 16 loci were discarded due to unspecific amplifications and 32 loci (14 tetranucleotide and 18 dinucleotide) were successfully scored. The average number of alleles per locus was 21 (±6.87, SD) and ranged from 8 to 34. The average observed and expected heterozygosities were 0.787 (±0.144 SD, range 0.250-0.935) and 0.909 (±0.122 SD, range 0.381-0.965), respectively. No significant linkage was detected. Eight loci showed deviations from Hardy-Weinberg equilibrium, and from these, four loci showed moderate null allele frequencies (0.104-0.220). The probability of individual identity for the new loci was 1.46. Genotyping error rates averaged 9.58%. The new markers will be useful to investigate patterns of larval dispersal, metapopulation dynamics, fine-scale genetic structure and diversity aimed to inform the implementation of spatially explicit fisheries management strategies in the Gulf of California.
我们为太平洋红鲷(Lutjanus peru)开发了一组高变微卫星标记,太平洋红鲷是墨西哥西海岸小规模渔业中一种具有重要经济价值的海洋鱼类。我们使用454 XL钛化学法进行了鸟枪法基因组测序,并利用生物信息学工具搜索完美的微卫星位点。我们选择了66对引物对,在来自加利福尼亚湾的32个个体中进行合成并在ABI PRISM 3730XL DNA测序仪上进行基因分型。我们估计了遗传多样性水平、连锁不平衡和哈迪 - 温伯格平衡的偏差,估计了无效等位基因的频率以及新标记的个体识别概率。我们对16个个体中的16个位点进行重新分析以估计基因分型错误率。18个位点未能扩增,16个位点因非特异性扩增而被舍弃,32个位点(14个四核苷酸和18个二核苷酸)成功分型。每个位点的平均等位基因数为21(±6.87,标准差),范围从8到34。观察到的平均杂合度和预期杂合度分别为0.787(±0.144标准差,范围0.250 - 0.935)和0.909(±0.122标准差,范围0.381 - 0.965)。未检测到显著连锁。8个位点显示出偏离哈迪 - 温伯格平衡,其中4个位点显示出中等的无效等位基因频率(0.104 - 0.220)。新位点的个体识别概率为1.46。基因分型错误率平均为9.58%。这些新标记将有助于研究幼体扩散模式、集合种群动态、精细尺度的遗传结构和多样性,旨在为加利福尼亚湾空间明确的渔业管理策略的实施提供信息。