Fusinatto L A, Lopes S, Silva-Ferreira A, Alexandrino J, Haddad C F B, Rocha C F D, Sequeira F
Departamento de Ecologia, Instituto de Biologia Roberto Alcântara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ, Brasil.
Genet Mol Res. 2013 Jan 24;12(1):230-4. doi: 10.4238/2013.January.24.15.
The new-generation 454 GS-FLX Titanium pyrosequencing was used to isolate microsatellite markers for the Brazilian Guanabara frog, Euparkerella brasiliensis, an Atlantic forest endemic species. Three multiplex polymerase chain reaction sets were optimized for genotyping of 11 polymorphic (di- and tetranucleotide) microsatellite markers. Genetic diversity was assessed in 21 individuals from a population (Reserva Ecológica de Guapiaçu, REGUA) located in the central region of the Rio de Janeiro State, in Brazil. The mean number of alleles per locus ranged from 3 to 12. Observed and expected heterozygosities ranged from 0.095 to 0.905 and from 0.094 to 0.904, respectively. After using the Bonferroni correction for multiple tests, there was no evidence of linkage disequilibrium between pairs of loci but deviations for Hardy-Weinberg equilibrium were found in 4 loci. We found no evidence for allele dropouts or stuttering, but we detected the presence of null alleles at loci Eb10 and Eb36. These markers will be useful for analyses of fine-scale population structure and determination of relative effects of habitat loss and fragmentation on population genetic variability within species.
新一代454 GS-FLX Titanium焦磷酸测序技术被用于为巴西瓜纳巴拉蛙（Euparkerella brasiliensis）分离微卫星标记，该蛙是大西洋森林的特有物种。优化了三个多重聚合酶链式反应组，用于对11个多态性（二核苷酸和四核苷酸）微卫星标记进行基因分型。对来自巴西里约热内卢州中部一个种群（瓜皮阿苏生态保护区，REGUA）的21个个体进行了遗传多样性评估。每个位点的等位基因平均数在3到12之间。观察到的杂合度和预期杂合度分别在0.095到0.905和0.094到0.904之间。在对多重检验使用邦费罗尼校正后，没有证据表明位点对之间存在连锁不平衡，但在4个位点发现了偏离哈迪-温伯格平衡的情况。我们没有发现等位基因缺失或结巴的证据，但在Eb10和Eb36位点检测到了无效等位基因的存在。这些标记将有助于分析精细尺度的种群结构，以及确定栖息地丧失和破碎化对物种内种群遗传变异性的相对影响。
