Abstract
The use of microsatellite markers provides a window into the evolutionary processes of a given species. As such, these markers are widely used in scientific and applied research and are praised for their practicality and ease of use; however, the unavoidable incidence of genotyping deviations has been broadly neglected in the literature. Therefore, the present study aimed to estimate the rate of null alleles, mutations, and genotyping errors in microsatellite loci, using Araucaria angustifolia, a threatened species, as a case study. We estimated the rates of the different types of genotyping deviations using mother-progeny genotype comparison from 50 seed-trees and their respective progeny (seeds). A total of 2336 A. angustifolia samples were genotyped, and we found that the rate of null alleles was 0.045. From the 1972 mother-progeny comparisons, the overall genotype deviation rate was 1.58%, consisting of 145 inconsistences (mutations), 339 null alleles, and 210 genotyping errors. In terms of seed numbers, 128 (6.5%) showed inconsistencies in at least one locus, 118 (6.0%) null alleles, and 321 (16.3%) genotyping errors. This is the first study to describe the inconsistences (mutations) between mother-progeny genotypes for A. angustifolia, and the outcome makes it clear that an understanding of these genotyping deviations must be considered in assessing the accuracy of inferences made based on population genetics analyses.
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Acknowledgments
We thank the Santa Catarina State University (UDESC) and Laboratório DNA UDESC, for logistical support, and, finally, landowners M. Duarte and J.A.R. Ribeiro for allowing access to their property.
Funding
We would like to thank the Fundação de Amparo a Pesquisa e Inovação do Estado de Santa Catarina (FAPESC) for providing financial support (project 14848/2011-2); the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for doctoral scholarships to N.C.F.C., M.B.L., and L.I.B.S. (Finance Code 001); and to the Conselho Nacional de Desenvolvimento Científico e Tecnológico—CNPq for the scholarship awarded to R.O.N.
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This research represents part of a Ph.D. thesis of N.C.F.C. that designed the research with the contribution of M.B.L., L.I.B.S. and R.O.N.; N.C.F.C. and L.I.B.S. collected the samples and performed the laboratory procedures; N.C.F.C. performed the analysis and wrote the draft of the manuscript. All authors have contributed towards manuscript writing, read and approved the final manuscript.
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The data archiving has been completed. We submitted the data to the TreeGenes Database and data accession is avaliable at https://doi.org/10.5281/zenodo.3406797.
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da Costa, N.C.F., Stedille, L.I.B., Lauterjung, M.B. et al. Distinguishing mutations and null alleles from genotyping errors using mother progeny comparisons in Brazilian pine (Araucaria angustifolia). Tree Genetics & Genomes 15, 78 (2019). https://doi.org/10.1007/s11295-019-1388-8
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DOI: https://doi.org/10.1007/s11295-019-1388-8