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DNA barcoding a taxonomically complex hemiparasitic genus reveals deep divergence between ploidy levels but lack of species-level resolution.

ABSTRACT: DNA barcoding is emerging as a useful tool not only for species identification but also for studying evolutionary and ecological processes. Although plant DNA barcodes do not always provide species-level resolution, the generation of large DNA barcode data sets can provide insights into the mechanisms underlying the generation of species diversity. Here, we study evolutionary processes in taxonomically complex British Euphrasia (Orobanchaceae), a group with multiple ploidy levels, frequent self-fertilization, young species divergence and widespread hybridization. We use a phylogenetic approach to investigate the colonization history of British Euphrasia, followed by a DNA barcoding survey and population genetic analyses to reveal the causes of shared sequence variation. Phylogenetic analysis shows Euphrasia have colonized Britain from mainland Europe on multiple occasions. DNA barcoding reveals that no British Euphrasia species has a consistent diagnostic sequence profile, and instead, plastid haplotypes are either widespread across species, or are population specific. The partitioning of nuclear genetic variation suggests differences in ploidy act as a barrier to gene exchange, while the divergence between diploid and tetraploid ITS sequences supports the polyploids being allotetraploid in origin. Overall, these results show that even when lacking species-level resolution, analyses of DNA barcoding data can reveal evolutionary patterns in taxonomically complex genera.

SUBMITTER: Wang X

PROVIDER: S-EPMC5941139 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature

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DNA barcoding a taxonomically complex hemiparasitic genus reveals deep divergence between ploidy levels but lack of species-level resolution.

Wang Xumei X Gussarova Galina G Ruhsam Markus M de Vere Natasha N Metherell Chris C Hollingsworth Peter M PM Twyford Alex D AD

AoB PLANTS 20180424 3

DNA barcoding is emerging as a useful tool not only for species identification but also for studying evolutionary and ecological processes. Although plant DNA barcodes do not always provide species-level resolution, the generation of large DNA barcode data sets can provide insights into the mechanisms underlying the generation of species diversity. Here, we study evolutionary processes in taxonomically complex British <i>Euphrasia</i> (Orobanchaceae), a group with multiple ploidy levels, frequen ...[more]

PMID: 29765588

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Project description:Premise of the studyThe internal transcribed spacer (ITS) region is situated between 18S and 26S in a polycistronic rRNA precursor transcript. It had been proved to be the most commonly sequenced region across plant species to resolve phylogenetic relationships ranging from shallow to deep taxonomic levels. Despite several taxonomical revisions in Cassiinae, a stable phylogeny remains elusive at the molecular level, particularly concerning the delineation of species in the genera Cassia, Senna and Chamaecrista. This study addresses the comparative potential of ITS datasets (ITS1, ITS2 and concatenated) in resolving the underlying morphological disparity in the highly complex genera, to assess their discriminatory power as potential barcode candidates in Cassiinae.MethodologyA combination of experimental data and an in-silico approach based on threshold genetic distances, sequence similarity based and hierarchical tree-based methods was performed to decipher the discriminating power of ITS datasets on 18 different species of Cassiinae complex. Lab-generated sequences were compared against those available in the GenBank using BLAST and were aligned through MUSCLE 3.8.31 and analysed in PAUP 4.0 and BEAST1.8 using parsimony ratchet, maximum likelihood and Bayesian inference (BI) methods of gene and species tree reconciliation with bootstrapping. DNA barcoding gap was realized based on the Kimura two-parameter distance model (K2P) in TaxonDNA and MEGA.Principal findingsBased on the K2P distance, significant divergences between the inter- and intra-specific genetic distances were observed, while the presence of a DNA barcoding gap was obvious. The ITS1 region efficiently identified 81.63% and 90% of species using TaxonDNA and BI methods, respectively. The PWG-distance method based on simple pairwise matching indicated the significance of ITS1 whereby highest number of variable (210) and informative sites (206) were obtained. The BI tree-based methods outperformed the similarity-based methods producing well-resolved phylogenetic trees with many nodes well supported by bootstrap analyses.ConclusionThe reticulated phylogenetic hypothesis using the ITS1 region mainly supported the relationship between the species of Cassiinae established by traditional morphological methods. The ITS1 region showed a higher discrimination power and desirable characteristics as compared to ITS2 and ITS1 + 2, thereby concluding to be the locus of choice. Considering the complexity of the group and the underlying biological ambiguities, the results presented here are encouraging for developing DNA barcoding as a useful tool for resolving taxonomical challenges in corroboration with morphological framework.

Project description:Accurate taxonomic identification of alien species is crucial to detect new incursions, prevent or reduce the arrival of new invaders and implement management options such as biological control. Globally, the taxonomy of non-native Prosopis species is problematic due to misidentification and extensive hybridization. We performed a genetic analysis on several Prosopis species, and their putative hybrids, including both native and non-native populations, with a special focus on Prosopis invasions in Eastern Africa (Ethiopia, Kenya and Tanzania). We aimed to clarify the taxonomic placement of non-native populations and to infer the introduction histories of Prosopis in Eastern Africa. DNA sequencing data from nuclear and chloroplast markers showed high homology (almost 100 %) between most species analysed. Analyses based on seven nuclear microsatellites confirmed weak population genetic structure among Prosopis species. Hybrids and polyploid individuals were recorded in both native and non-native populations. Invasive genotypes of Prosopis juliflora in Kenya and Ethiopia could have a similar native Mexican origin, while Tanzanian genotypes likely are from a different source. Native Peruvian Prosopis pallida genotypes showed high similarity with non-invasive genotypes from Kenya. Levels of introduced genetic diversity, relative to native populations, suggest that multiple introductions of P. juliflora and P. pallida occurred in Eastern Africa. Polyploidy may explain the successful invasion of P. juliflora in Eastern Africa. The polyploid P. juliflora was highly differentiated from the rest of the (diploid) species within the genus. The lack of genetic differentiation between most diploid species in their native ranges supports the notion that hybridization between allopatric species may occur frequently when they are co-introduced into non-native areas. For regulatory purposes, we propose to treat diploid Prosopis taxa from the Americas as a single taxonomic unit in non-native ranges.

Dataset Information

DNA barcoding a taxonomically complex hemiparasitic genus reveals deep divergence between ploidy levels but lack of species-level resolution.

Publications

DNA barcoding a taxonomically complex hemiparasitic genus reveals deep divergence between ploidy levels but lack of species-level resolution.

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