Search bioRxiv⌕ Search

bioRxiv · 10.1101/155861

Multilocus DNA barcoding - Species Identification with Multilocus Data

Abstract

Species identification using DNA sequences, known as DNA barcoding has been widely used in many applied fields. Current barcoding methods are usually based on a single mitochondrial locus, such as cytochrome c oxidase subunit I (COI). This type of barcoding is not always effective when applied to species separated by short divergence times or that contain introgressed genes from closely related species. Herein we introduce a more effective multi-locus barcoding framework that is based on gene capture and \"next-generation\" sequencing and provide both empirical and simulation tests of its efficacy. We examine genetic distinctness in two pairs of fishes that are sister-species: Siniperca chuatsi vs. S. kneri and Sicydium altum vs. S. adelum, where the COI barcoding approach failed species identification in both cases. Results revealed that distinctness between S. chuatsi and S. kneri increased as more independent loci were added. By contrast S. altum and S. adelum could not be distinguished even with all loci. Analyses of population structure and gene flow suggested that the two species of Siniperca diverged from each other a long time ago but have unidirectional gene flow, whereas the two species of Sicydium are not separated from each other and have high bidirectional gene flow. Simulations demonstrate that under limited gene flow (< 0.00001 per gene per generation) and enough separation time (> 100000 generation), we can correctly identify species using more than 90 loci. Finally, we selected 500 independent nuclear markers for ray-finned fishes and designed a three-step pipeline for multilocus DNA barcoding.

Source connections

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Liu, J., Jiang, J., Song, S., Tornabene, L., Chabarria, R., Naylor, G. J., Li, C.. 2017-06-27. Multilocus DNA barcoding - Species Identification with Multilocus Data. https://doi.org/10.1101/155861

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Integrative taxonomy of the North American pink flat-backed millipedes (Polydesmidae: Pseudopolydesmus Attems, 1898)

Millipedes of the genus Pseudopolydesmus Attems, 1898 are common millipedes in North America. They are the largest, in terms of body size, and most geographically wide-ranging of the family Polydesmidae, and are ubiquitous in eastern and central North American forests. Eight species have been named based on the morphology of the genitalia, but these have not been investigated genetically. Here, we take an integrated taxonomic approach, using five genes to infer a molecular phylogeny of Pseudopolydesmus species, and analyze a dataset of 15 morphological characters in an effort to delimit species in the genus. 1,148 Pseudopolydesmus specimens were newly collected for this study, and all known species were sampled. These specimens were used for analysis, and were combined with museum specimens and literature records into a database of 3,462 unique specimen records. This database was used to produce distribution maps for each species in the genus. The maximum likelihood phylogeny found support for the eight species in the genus, supporting previous morphological species hypotheses. However, previous morphology-based hypotheses regarding evolutionary relationships between the species were not supported by the analysis. The morphological analysis, performed with a modified PCA, did not delimit species clusters as well as the genitalic and genetic differences did, due to comparatively uniform somatic morphology in the genus. However, our results show that genitalic morphology, combined with genetic analysis, provide congruent species delimitation hypotheses for Pseudopolydesmus, and these lines of evidence may prove useful in delimiting species and generic boundaries in other Polydesmidae worldwide.

evolutionary biology↗

The pupal stage shapes life-history trade-offs across metamorphosis

Dealing with ecological stressors during development often comes at the expense of later-life performance. Although such trade-offs are well studied as two-stage systems such as larva-adult trade-offs, they are rarely examined across multiple developmental stages, limiting our understanding of how ecological effects are integrated throughout complex life cycles. We addressed this gap using a multistage experimental framework in Aedes aegypti, where we exposed individuals to predation risk during the larval stage, the pupal stage, both stages, or neither, and quantified developmental, physiological, and adult fitness traits. Contrary to expectations, predation risk across successive stages did not produce cumulative fitness costs, and traits varied in their responses. Both males and females emerged faster when exposed to risk, with lighter bodies, but males paid an additional cost of reduced starvation resistance when the pupal stage was exposed. Surprisingly, despite their lower body mass, females showed enhanced reproductive performance when both stages were exposed to risk. Overall, responses were stage- and sex-specific, with exposure during the non-feeding pupal stage imposing greater life-history costs than larval exposure alone. Measurements across the larva-pupa-adult continuum further revealed that traits such as body mass and energy reserves respond differently before and after metamorphosis, highlighting the role of the pupa in mediating trade-offs. Our findings identify the pupal stage as a critical mediator of life-history trade-offs and demonstrate the value of a multistage framework for understanding the role of each stage in life-history evolution.

evolutionary biology↗

Cell-lineage selection interacts with development and life history to shape the rates and identity of heritable mutations

Mutation rates determine the supply of heritable variation and are central to evolutionary theory, yet they are usually treated as properties of molecular mutational processes. We hypothesised that mutation rates are also shaped by selection among cell lineages within organisms and that the strength of this process depends on developmental and life-history traits. To test the extent of this idea, we combined mathematical modelling with experiments in Drosophila melanogaster and treated mutation rates as emergent properties of interactions among molecular mutagenesis, development, life history, and cell-lineage selection. The model predicts that even modest effects of mutations on cellular fitness can substantially alter the probability that mutations are transmitted, and that these effects depend strongly on developmental architecture, germline dynamics, and time to reproduction. In Drosophila, we experimentally demonstrate biased transmission of a mutation affecting cellular proliferation and show that this bias changes over the reproductive lifespan of the individual. Extending the model genome-wide reveals that the cellular distribution of fitness effects and its interaction with developmental parameters influence both the estimated mutation rate and the composition of transmitted mutations, producing transmission biases that favour mutations with particular cellular fitness effects. These results suggest that developmental and life-history parameters determine how cellular fitness effects translate into transmission bias; therefore, estimates of heritable mutational input can vary from species to species even under constant molecular mutagenesis. By filtering mutations before they enter populations, developmental and life-history evolution can modify both the rate and the nature of heritable variation and thereby influence evolutionary trajectories.

evolutionary biology↗