A test of psbK-psbI and atpF-atpH as potential plant DNA barcodes using the flora of the Kruger National Park as a model system (South Africa)

A DNA barcode consists of a standardized short sequence of DNA (400-800bp) used to identify the taxonomic species a small organic fragment belongs to. Even though it has been easy to discriminate animal species by using the mitochondrial gene cox1, this is still difficult for plants seeing that the mitochondrial genome is not variable enough on the species level. During the Second International Barcode of Life Conference in Tapei (September 2007), different plastid regions were proposed as potential plant DNA barcodes, such as atpF-atpH and psbK-psbI, but no consensus on which region to use was reached during the meeting. The largest plant DNA barcoding study to date proposed matK as the best candidate and suggested that in combination with trnH-psbA a slight increase in performance could be achieved. However, no study has tested the suitability of the newly proposed psbK-psbI and atpF-atpH for plant barcoding purposes. Four potential DNA barcodes, matK, trnH-psbA, atpF-atpH, and psbK-psbI, were amplified and sequenced for a selective sampling including mainly trees and shrubs of the flora of the Kruger National Park Africa (South Africa). The performance of each region and also each possible combination of these were tested by applying a battery of metrics and statistical tests. Our results confirm that the second half (5’ end) of matK is the best candidate in a single locus barcoding approach reaching 87.5% of species correctly identified. Combining matK with trnH-psbA and psbK-psbI increased only slightly the performance in discriminating species. The results from this study show that the use of a ‘three-region barcode’ does not significantly outperform matK in a single-locus barcoding approach. We therefore argue against the ‘multiple barcode approach’ proposed by the plant working group, and instead propose to keep barcoding plants in line with the approach taken for animals, i.e. using one barcode: cox1 for animals and matK for plants

Evidence of constant diversification punctuated by a mass extinction in the African cycads

The recent evidence that extant cycads are not living fossils triggered a renewed search for a better understanding of their evolutionary history. In this study, we investigated the evolutionary diversification history of the genus Encephalartos, a monophyletic cycad endemic to Africa. We found an antisigmoidal pattern with a plateau and punctual explosive radiation. This pattern is typical of a constant radiation with mass extinction. The rate shift that we found may therefore be a result of a rapid recolonization of niches that have been emptied owing to mass extinction. Because the explosive radiation occurred during the transition Pliocene–Pleistocene, we argued that the processes might have been climatically mediated

Phylogeny of Iridaceae Subfamily Crocoideae Based on a Combined Multigene Plastid DNA Analysis

The phylogeny of Crocoideae, the largest of four subfamilies currently recognized in Tridaceae, has eluded resolution until sequences of two more plastid DNA regions were added here to a previously published matrix containing sequences from four DNA plastid regions. Sister to the core Nivenioideae, the woody Klattia, Nivenia, and Witsenia, Crocoideae are a climax group in lridaceae, comprising some 995 species, slightly more than half of the total in the family. Synapomorphies of Crocoideae include pollen exine perforate, pollen aperture operculate, ovule campylotropous (or hypotropous), root xylem vessels with simple perforations, cormous rootstock, inflorescence a spike, and plants deciduous. The six DNA region analysis here that includes examples of 27 of the 28 genera of the subfamily shows the southern African Tritoniopsis sister to the remaining genera, which resolve into four well-supported clusters (bootstrap support \u3e85%). Each of these major clades is treated as a tribe, the synapomorphies of which are discussed in light of the molecular phylogenetic analyses. Original embryological and seed developmental studies largely support the tribal classification. Tritoniopsideae alone has the inner floral bracts not forked apically, and a hypotropous ovule, while this tribe and Watsonieae have axillary corm development. The remaining three tribes have apical corm development, and together with Watsonieae have a campylotropous ovule, and the inner layer of the inner integument crushed at maturity

Unravelling the evolutionary origins of biogeographic assemblages

AIM : Floristic and faunal diversity fall within species assemblages that can be grouped into distinct biomes or ecoregions. Understanding the origins of such biogeographic assemblages helps illuminate the processes shaping present‐day diversity patterns and identifies regions with unique or distinct histories. While the fossil record is often sparse, dated phylogenies can provide a window into the evolutionary past of these regions. Here, we present a novel phylogenetic approach to investigate the evolutionary origins of present‐day biogeographic assemblages and highlight their conservation value. LOCATION : Southern Africa. METHODS : We evaluate the evolutionary turnover separating species clusters in space at different time slices to determine the phylogenetic depth at which the signal for their present‐day structure emerges. We suggest present‐day assemblages with distinct evolutionary histories might represent important units for conservation. We apply our method to the vegetation of southern Africa using a dated phylogeny of the woody flora of the region and explore how the evolutionary history of vegetation types compares to common conservation currencies, including species richness, endemism and threat. RESULTS : We show the differentiation of most present‐day vegetation types can be traced back to evolutionary splits in the Miocene. The woody flora of the Fynbos is the most evolutionarily distinct, and thus has deeper evolutionary roots, whereas the Savanna and Miombo Woodland show close phylogenetic affinities and likely represent a more recent separation. However, evolutionarily distinct phyloregions do not necessarily capture the most unique phylogenetic diversity, nor are they the most species‐rich or threatened. MAIN CONCLUSIONS : Our approach complements analyses of the fossil record and serves as a link to the history of diversification, migration and extinction of lineages within biogeographic assemblages that is separate from patterns of species richness and endemism. Our analysis reveals how phyloregions capture conservation value not represented by traditional biodiversity metrics.The Government of Canada through Genome Canada and Ontario Genomics Institute (2008‐OGI‐ICI‐03), International Development Research Centre (IDRC) and University of Johannesburg.http://wileyonlinelibrary.com/journal/ddi2019-03-01hj2018Plant Production and Soil Scienc

Molecular and morphological analysis of subfamily Alooideae (Asphodelaceae) and the inclusion of chortolirion in aloe

Asphodelaceae subfam. Alooideae (Asparagales) currently comprises five genera, four of which are endemic to southern Africa. Despite their importance in commercial horticulture the evolutionary relationships among the genera are still incompletely understood. This study examines phylogenetic relationships in the subfamily using an expanded molecular sequence dataset from three plastid regions (matK, rbcLa, trnH-psbA) and the first subunit of the nuclear ribosomal internal transcribed spacer (ITS1). Sequence data were analysed using maximum parsimony and Bayesian statistics, and selected morphological traits were mapped onto the molecular phylogeny. Haworthia is confirmed as being polyphyletic, comprising three main clades that largely correlate with current subgeneric circumscriptions. Astroloba and Gasteria are evidently each monophyletic and sister respectively to Astroloba and H. subg. Robustipedunculares. Chortolirion is shown to be deeply nested within Aloe and is formally included in that genus. Aloe itself is clearly polyphyletic, with the dwarf species A. aristata allied to Haworthia subg. Robustipedunculares. The taxonomic implications of these findings are examined but branch support at critical lower nodes is insufficient at this stage to justify implementing major taxonomic changes

Bacterial diversity in the waterholes of the Kruger National Park : an eDNA metabarcoding approach

Bacteria are essential components of natural environments. They contribute to ecosystem functioning through roles as mutualists and pathogens for larger species, and as key components of food webs and nutrient cycles. Bacterial communities respond to environmental disturbances, and the tracking of these communities across space and time may serve as indicators of ecosystem health in areas of conservation concern. Recent advances in DNA sequencing of environmental samples allow for rapid and culture-free characterization of bacterial communities. Here we conduct the first metabarcoding survey of bacterial diversity in the waterholes of the Kruger National Park, South Africa. We show that eDNA can be amplified from waterholes and find strongly structured microbial communities, likely reflecting local abiotic conditions, animal ecology, and anthropogenic disturbance. Over timescales from days to weeks we find increased turnover in community composition, indicating bacteria may represent host-associated taxa of large vertebrates visiting the waterholes. Through taxonomic annotation we also identify pathogenic taxa, demonstrating the utility of eDNA metabarcoding for surveillance of infectious diseases. These samples serve as a baseline survey of bacterial diversity in the Kruger National Park, and in the future, spatially distinct microbial communities may be used as markers of ecosystem disturbance, or biotic homogenization across the park.This paper is part of a special issue entitled “Trends in DNA Barcoding and Metabarcoding”.M.J.F. was supported by a Vanier NSERC CGS and the CIHR Systems Biology Training Program, with project funding supported by the Quebec Centre for Biodiversity Science, the McGill Biology Department, and an NSERC Discovery Grant awarded to T.J.D.http://www.nrcresearchpress.com/journal/genhj2019Paraclinical Science

Biodiversity baselines : tracking insects in Kruger National Park with DNA barcodes

Reflecting their species richness and ecological diversification, insects play a central role in terrestrial ecosystems but difficulties in species-level assignments have restricted large-scale analysis of their community structure. Employing South Africa’s largest national park as a model system, we demonstrate that DNA barcoding can break this barrier. A year-long deployment of Malaise traps at 25 sites in Kruger National Park (KNP) generated 1000+ weekly collections containing about 800,000 specimens. Insect biomass averaged 1.05 g per trap-day but varied by up to 2-fold between months, being lower in the dry than wet season. Nearly 370,000 specimens were individually analyzed to reveal 19,730 Barcode Index Numbers (BINs; species proxy), a count equal to 43% of the known insect fauna of southern Africa. There was clear differentiation in insect richness and composition between KNP’s two ecoregions, but little among the vegetation types comprising them. The spatial gradient in annual rainfall explained more than half of the variation in compositional similarity among sites with less similarity among samples in the wet season, particularly among those in high rainfall areas. These results suggest that the factors organising insect communities in KNP are not fine-scale vegetation differences, but larger-scale processes associated with ecoregions and rainfall. Estimates of sample coverage indicate that the species not collected are rare, comprising only 4% of the individuals in the community. With a well-parameterized DNA barcode reference library in place, metabarcoding can be used to assess future shifts in the insect fauna of KNP rapidly and inexpensively.http://www.elsevier.com/locate/bioconpm2021Paraclinical Science

A novel phylogenetic regionalization of phytogeographical zones of southern Africa reveals their hidden evolutionary affinities

AIM : Although existing bioregional classification schemes often consider the compositional affinities within regional biotas, they do not typically incorporate phylogenetic information explicitly. Because phylogeny captures information on the evolutionary history of taxa, it provides a powerful tool for delineating biogeographical boundaries and for establishing relationships among them. Here, we present the first vegetation delineation of the woody flora of southern Africa based upon evolutionary relationships. LOCATION : Southern Africa. METHODS : We used a published time-calibrated phylogenetic tree for 1400 woody plant species along with their geographical distributions and a metric of phylogenetic beta diversity to generate a phylogenetic delineation of the woody vegetation of southern Africa. We then explored environmental correlates of phylogenetic turnover between them, and the evolutionary distinctiveness of the taxa within them. RESULTS : We identified 15 phylogenetically distinct biogeographical units, here referred to as phyloregions. The largest phyloregion broadly overlaps with Savanna vegetation, while the phyloregion overlapping with the south-western portion of the Fynbos biome is the most evolutionarily distinct. Potential evapotranspiration and mean annual temperature differ significantly among phyloregions and correlate with patterns of phylogenetic beta diversity between them. Our phylogeny-based delimitation of southern Africa’s woody vegetation broadly matches currently recognized phytogeographical classifications, but also highlights parts of the Namib Karoo and Greater Limpopo Transfrontier Park as distinct, but previously under-recognized biogeographical units. MAIN CONCLUSIONS : Our analysis provides new insights into the structure and phylogenetic relationships among the woody flora of southern Africa. We show that evolutionary affinities differentiate phyloregions closely resembling existing vegetation classifications, yet also identify ‘cryptic’ phyloregions that are as evolutionarily distinct as some of the recognized African vegetation types.Government of Canada through Genome Canada and the Ontario Genomics Institute (2008-OGI-ICI-03), the International Development Research Centre (IDRC) Canada, the University of Johannesburg and the South African National Research Foundation (NRF).http://onlinelibrary.wiley.com/doi/10.1111/jbi.126192017-01-31hb201

DNA barcodes reveal microevolutionary signals in fire response trait in two legume genera

Large-scale DNA barcoding provides a new technique for species identification and evaluation of relationships across various levels (populations and species) and may reveal fundamental processes in recently diverged species. Here, we analysed DNA sequence variation in the recently diverged legumes from the Psoraleeae (Fabaceae) occurring in the Cape Floristic Region (CFR) of southern Africa to test the utility of DNA barcodes in species identification and discrimination. We further explored the phylogenetic signal on fire response trait (reseeding and resprouting) at species and generic levels. We showed that Psoraleoid legumes of the CFR exhibit a barcoding gap yielding the combination of matK and rbcLa (matK + rbcLa) dataset as a better barcode than single regions. We found a high score (100%) of correct identification of individuals to their respective genera but very low score (<50%) in identifying them to species. We found a considerable match (54%) between genetic species and morphologicallydelimited species. We also found that different lineages showed a weak but significant phylogenetic conservatism in their response to fire as reseeders or resprouters, with more clustering of resprouters than would be expected by chance. These novel microevolutionary patterns might be acting continuously over time to produce multi-scale regularities of biodiversity. This study provides the first insight into the DNA barcoding campaign of land plants in species identification and detection of phylogenetic signal in recently diverged lineages of the CFR.The South African National Research Foundation (NRF; AMM); Nigeria Tertiary Education Trust Fund (NTETF) / Umaru Musa Yar’adua University Katsina, Nigeria (Fellowship Grant; A. Bello); and University of Cape Town, J. W. Jagger Centenary Gift Scholarship (to A. Bello).http://aobpla.oxfordjournals.orgam2016Physiotherap

Telling plant species apart with DNA:From barcodes to genomes

Land plants underpin a multitude of ecosystem functions, support human livelihoods and represent a critically important component of terrestrial biodiversity—yet many tens of thousands of species await discovery, and plant identification remains a substantial challenge, especially where material is juvenile, fragmented or processed. In this opinion article, we tackle two main topics. Firstly, we provide a short summary of the strengths and limitations of plant DNA barcoding for addressing these issues. Secondly, we discuss options for enhancing current plant barcodes, focusing on increasing discriminatory power via either gene capture of nuclear markers or genome skimming. The former has the advantage of establishing a defined set of target loci maximizing efficiency of sequencing effort, data storage and analysis. The challenge is developing a probe set for large numbers of nuclear markers that works over sufficient phylogenetic breadth. Genome skimming has the advantage of using existing protocols and being backward compatible with existing barcodes; and the depth of sequence coverage can be increased as sequencing costs fall. Its non-targeted nature does, however, present a major informatics challenge for upscaling to large sample sets. This article is part of the themed issue ‘From DNA barcodes to biomes’