Characterization of 14 novel microsatellite loci in the Argentine black and white tegu (Salvator merianae) via 454 pyrosequencing

We have developed 14 novel microsatellite loci for the Argentine black and white tegu (Salvator merianae), using samples from invasive populations from the US state of Florida. Pyrosequencing was used to identify 3154 potentially amplifiable microsatellite loci and to subsequently develop 14 informative markers. These 14 markers were screened for variation in 40 individuals from Miami-Dade County, Florida. All loci were polymorphic and contained between 2 and 4 alleles per locus (mean ± SE = 2.71 ± 0.24), with observed heterozygosity ranging from 0.00 to 0.88 (mean ± SE = 0.38 ± 0.07). Four of the loci (Teg4, Teg5, Teg17, Teg19) significantly deviated from Hardy-Weinberg proportions and three of these loci (Teg4, Teg5, Teg19) showed evidence of null alleles. In addition, there was statistical evidence for genotypic disequilibrium between Teg14 and Teg19. BLASTn searches of NCBI’s ‘nr/nt’ database using microsatellite containing 454 fragments as queries were largely uninformative; however, it is likely that some of these markers will be of utility in S. merinae’s native range.

Supplementary Data File 1: Zip archive containing FASTA files of the 454 sequences generated during the course of our study.Supplementary Data File 2: Excel spreadsheet containing the primers associated with the potentially amplifiable loci identified during the course of our study.

本研究针对阿根廷黑白鬣蜥（Salvator merianae）成功构建了14个新型微卫星标记位点，这些位点由美国佛罗里达州入侵种群样本提供。通过焦磷酸测序技术，我们识别出3154个潜在的扩增微卫星位点，并在此基础上开发出14个具有信息量的标记。这些标记在佛罗里达州迈阿密-戴德县的40个个体中进行了变异筛选。所有标记位点均呈现多态性，每个位点包含2至4个等位基因（平均±标准误=2.71±0.24），观察到的杂合度介于0.00至0.88之间（平均±标准误=0.38±0.07）。其中，四个位点（Teg4、Teg5、Teg17、Teg19）与哈迪-温伯格比例存在显著差异，且其中三个位点（Teg4、Teg5、Teg19）显示出缺失等位基因的证据。此外，Teg14与Teg19之间还存在着基因型不平衡的统计证据。利用含有454片段的微卫星序列作为查询，在NCBI的‘nr/nt’数据库中进行BLASTn搜索，结果大多缺乏信息；然而，预计这些标记中的一些将在S. merinae的原生区域内具有实际应用价值。

补充数据文件1：包含本研究过程中生成的454序列的FASTA文件压缩包。

补充数据文件2：包含本研究过程中识别的潜在可扩增位点相关引物的Excel电子表格。