Use of DNA barcodes to identify flowering plants
07 Jun 2005-Proceedings of the National Academy of Sciences of the United States of America (National Academy of Sciences)-Vol. 102, Iss: 23, pp 8369-8374
TL;DR: Comparison of the total plastid genomes of tobacco and deadly nightshade enhanced with trials on widely divergent angiosperm taxa suggest that the sequences in this pair of loci have the potential to discriminate among the largest number of plant species for barcoding purposes.
Abstract: Methods for identifying species by using short orthologous DNA sequences, known as “DNA barcodes,” have been proposed and initiated to facilitate biodiversity studies, identify juveniles, associate sexes, and enhance forensic analyses. The cytochrome c oxidase 1 sequence, which has been found to be widely applicable in animal barcoding, is not appropriate for most species of plants because of a much slower rate of cytochrome c oxidase 1 gene evolution in higher plants than in animals. We therefore propose the nuclear internal transcribed spacer region and the plastid trnH-psbA intergenic spacer as potentially usable DNA regions for applying barcoding to flowering plants. The internal transcribed spacer is the most commonly sequenced locus used in plant phylogenetic investigations at the species level and shows high levels of interspecific divergence. The trnH-psbA spacer, although short (≈450-bp), is the most variable plastid region in angiosperms and is easily amplified across a broad range of land plants. Comparison of the total plastid genomes of tobacco and deadly nightshade enhanced with trials on widely divergent angiosperm taxa, including closely related species in seven plant families and a group of species sampled from a local flora encompassing 50 plant families (for a total of 99 species, 80 genera, and 53 families), suggest that the sequences in this pair of loci have the potential to discriminate among the largest number of plant species for barcoding purposes.
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Royal Botanic Garden Edinburgh1, National Institutes of Health2, University of Guelph3, University of Johannesburg4, Royal Botanic Gardens5, Smithsonian Institution6, University of British Columbia7, Natural History Museum8, Korea University9, University of Toronto10, State University of Feira de Santana11, University of Costa Rica12, Columbus State University13, University of Wisconsin-Madison14, New York Botanical Garden15, University of the Andes16, University of Cape Town17, Seoul National University18, Hallym University19, National Autonomous University of Mexico20, Imperial College London21
TL;DR: The 2-locus combination of rbcL+matK will provide a universal framework for the routine use of DNA sequence data to identify specimens and contribute toward the discovery of overlooked species of land plants.
Abstract: DNA barcoding involves sequencing a standard region of DNA as a tool for species identification. However, there has been no agreement on which region(s) should be used for barcoding land plants. To provide a community recommendation on a standard plant barcode, we have compared the performance of 7 leading candidate plastid DNA regions (atpF–atpH spacer, matK gene, rbcL gene, rpoB gene, rpoC1 gene, psbK–psbI spacer, and trnH–psbA spacer). Based on assessments of recoverability, sequence quality, and levels of species discrimination, we recommend the 2-locus combination of rbcL+matK as the plant barcode. This core 2-locus barcode will provide a universal framework for the routine use of DNA sequence data to identify specimens and contribute toward the discovery of overlooked species of land plants.
2,255 citations
TL;DR: Nine newly explored regions of the chloroplast genome offer levels of variation better than the best regions identified in an earlier study and are therefore likely to be the best choices for molecular studies at low taxonomic levels.
Abstract: Although the chloroplast genome contains many noncoding regions, relatively few have been exploited for interspecific phylogenetic and intraspecific phylogeographic studies. In our recent evaluation of the phylogenetic utility of 21 noncoding chloroplast regions, we found the most widely used noncoding regions are among the least variable, but the more variable regions have rarely been employed. That study led us to conclude that there may be unexplored regions of the chloroplast genome that have even higher relative levels of variability. To explore the potential variability of previously unexplored regions, we compared three pairs of single-copy chloroplast genome sequences in three disparate angiosperm lineages: Atropa vs. Nicotiana (asterids); Lotus vs. Medicago (rosids); and Saccharum vs. Oryza (monocots). These three separate sequence alignments highlighted 13 mutational hotspots that may be more variable than the best regions of our former study. These 13 regions were then selected for a more detailed analysis. Here we show that nine of these newly explored regions (rpl32-trnL((UAG)), trnQ((UUG))-5'rps16, 3'trnV((UAC))-ndhC, ndhF-rpl32, psbD-trnT((GGU)), psbJ-petA, 3'rps16-5'trnK((UUU)), atpI-atpH, and petL-psbE) offer levels of variation better than the best regions identified in our earlier study and are therefore likely to be the best choices for molecular studies at low taxonomic levels.
1,840 citations
Cites background or methods from "Use of DNA barcodes to identify flo..."
...Lastly, Kress et al. (2005) compared the complete chloroplast genomes of Atropa and Nicotiana to find an appropriate region for DNA barcoding in plants....
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...In the Kress et al. (2005), Daniell et al. (2006), and Timme et al. (2007) papers, the authors compiled a list of the most variable (on a percentage basis) noncoding regions of the chloroplast genome....
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...The mostly widely used is to calculate percentage variability as was done by Kress et al. (2005), Daniell et al. (2006), and Timme et al. (2007)....
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...Both Kress et al. (2005) and Rubinoff et al. (2006) suggest that in all likelihood we will need to employ more than one marker in a barcoding effort, and we feel that our work provides the necessary background to make an informed decision as to which cpDNA markers might be candidates for such an…...
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...Kress et al. (2005) suggested that the trnH-psbA intergenic spacer is a useful marker for such an effort —and we agree that this may be about as good a marker as there is in the chloroplast genome for three reasons: (1) the trnH-psbA intergenic spacer is among the most variable (in terms of…...
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TL;DR: The second internal transcribed spacer (ITS2) of nuclear ribosomal DNA represents the most suitable region for DNA barcoding applications and can be potentially used as a standard DNA barcode to identify medicinal plants and their closely related species.
Abstract: Background
The plant working group of the Consortium for the Barcode of Life recommended the two-locus combination of rbcL + matK as the plant barcode, yet the combination was shown to successfully discriminate among 907 samples from 550 species at the species level with a probability of 72%. The group admits that the two-locus barcode is far from perfect due to the low identification rate, and the search is not over.
1,252 citations
Cites background or methods or result from "Use of DNA barcodes to identify flo..."
...In previous studies, most researchers accepted psbA-trnH as a potential plant barcode [9,15,16,18,19,21,33]....
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...8S, the internal transcribed spacer (ITS) of nuclear ribosomal DNA and regions of the ITS could be potential barcodes [9,14] (Fig....
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...The sequences of the universal primers for the DNA barcode to be tested, including those for psbA-trnH, matK, rbcL, rpoC1, ycf5 and ITS, and general PCR reaction conditions were obtained from previous studies [9,17,18,21]....
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...First, most single-copy genes in the nuclear genome, as well as their introns, have been excluded as barcode candidates because of the lack of universal primers for their amplification [9]....
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...[9] compared 10 loci for authenticating closely related species in 7 plant families and 99 species belonging to 88 genera in 53 families, and they reported that the psbA-trnH spacer and the...
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TL;DR: Despite the potential benefits of DNA barcoding to both the practitioners and users of taxonomy, it has been controversial in some scientific circles and a few have even characterized it as being “anti-taxonomy,” arguing that its implementation will signal the death of a system 250 years in the making.
Abstract: DNA barcoding is a novel system designed to provide rapid, accurate, and automatable species identifications by using short, standardized gene regions as internal species tags. As a consequence, it will make the Linnaean taxonomic system more accessible, with benefits to ecologists, conservationists, and the diversity of agencies charged with the control of pests, invasive species, and food safety. More broadly, DNA barcoding allows a day to be envisioned when every curious mind, from professional biologists to schoolchildren, will have easy access to the names and biological attributes of any species on the planet. In addition to assigning specimens to known species, DNA barcoding will accelerate the pace of species discovery by allowing taxonomists to rapidly sort specimens and by highlighting divergent taxa that may represent new species. By augmenting their capabilities in these ways, DNA barcoding offers taxonomists the opportunity to greatly expand, and eventually complete, a global inventory of life’s diversity. Despite the potential benefits of DNA barcoding to both the practitioners and users of taxonomy, it has been controversial in some scientific circles (Wheeler, 2004; Will and Rubinoff, 2004; Ebach and Holdredge, 2005; Will et al., 2005). A few have even characterized DNA barcoding as being “anti-taxonomy,” arguing that its implementation will signal the death of a system 250 years in the making. We feel that this opposition stems from misconceptions about the DNA barcoding effort. As such, we welcome this opportunity to clarify both the rationale and potential impacts of DNA barcoding. In responding to this set of questions, we emphasize the multiple positive impacts of this approach for taxonomy and biodiversity science.
1,148 citations
Cites background from "Use of DNA barcodes to identify flo..."
...The core principles of barcode analysis (minimalization and standardization of sequence targets) are surely applicable to these organisms, but the selection of gene regions and tests of their effectiveness remain in progress although early results on plants (Kress et al., 2005) and protists (G....
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...…standardization of sequence targets) are surely applicable to these organisms, but the selection of gene regions and tests of their effectiveness remain in progress although early results on plants (Kress et al., 2005) and protists (G.W. Saunders, personal communication) provide cause for optimism....
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TL;DR: A combination of the non-coding trnH-psbA spacer region and a portion of the coding rbcL gene is recommended as a two-locus global land plant barcode that provides the necessary universality and species discrimination.
Abstract: Background. A useful DNA barcode requires sufficient sequence variation to distinguish between species and ease of application across a broad range of taxa. Discovery of a DNA barcode for land plants has been limited by intrinsically lower rates of sequence evolution in plant genomes than that observed in animals. This low rate has complicated the trade-off in finding a locus that is universal and readily sequenced and has sufficiently high sequence divergence at the species-level. Methodology/Principal Findings. Here, a global plant DNA barcode system is evaluated by comparing universal application and degree of sequence divergence for nine putative barcode loci, including coding and non-coding regions, singly and in pairs across a phylogenetically diverse set of 48 genera (two species per genus). No single locus could discriminate among species in a pair in more than 79% of genera, whereas discrimination increased to nearly 88% when the non-coding trnH-psbA spacer was paired with one of three coding loci, including rbcL. In silico trials were conducted in which DNA sequences from GenBank were used to further evaluate the discriminatory power of a subset of these loci. These trials supported the earlier observation that trnH-psbA coupled with rbcL can correctly identify and discriminate among related species. Conclusions/ Significance. A combination of the non-coding trnH-psbA spacer region and a portion of the coding rbcL gene is recommended as a two-locus global land plant barcode that provides the necessary universality and species discrimination. Citation: Kress WJ, Erickson DL (2007) A Two-Locus Global DNA Barcode for Land Plants: The Coding rbcL Gene Complements the Non-Coding trnH
1,088 citations
Cites background or methods from "Use of DNA barcodes to identify flo..."
...for flowering plants [8], in this study proved less favorable because of the low primer success across land plants (60....
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...The trnH-psbA spacer was treated according to Kress, Wurdack, Zimmer, Weigt and Janzen [8]....
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...The selection of a barcode locus is, however, complicated by the trade-off that arises between the need for universal application and maximal rates of sequence divergence [8]....
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...For flowering plants the non-coding plastid trnH-psbA intergenic spacer region and the multicopy nuclear Internal Transcribed Spacer (ITS) are two of the leading candidates [8]....
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...DISCUSSION The results suggest that the non-coding trnH-psbA intergenic spacer remains the most viable candidate for a single-locus barcode for land plants [8]....
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13,139 citations
TL;DR: It is established that the mitochondrial gene cytochrome c oxidase I (COI) can serve as the core of a global bioidentification system for animals and will provide a reliable, cost–effective and accessible solution to the current problem of species identification.
Abstract: Although much biological research depends upon species diagnoses, taxonomic expertise is collapsing. We are convinced that the sole prospect for a sustainable identification capability lies in the construction of systems that employ DNA sequences as taxon 'barcodes'. We establish that the mitochondrial gene cytochrome c oxidase I (COI) can serve as the core of a global bioidentification system for animals. First, we demonstrate that COI profiles, derived from the low-density sampling of higher taxonomic categories, ordinarily assign newly analysed taxa to the appropriate phylum or order. Second, we demonstrate that species-level assignments can be obtained by creating comprehensive COI profiles. A model COI profile, based upon the analysis of a single individual from each of 200 closely allied species of lepidopterans, was 100% successful in correctly identifying subsequent specimens. When fully developed, a COI identification system will provide a reliable, cost-effective and accessible solution to the current problem of species identification. Its assembly will also generate important new insights into the diversification of life and the rules of molecular evolution.
9,879 citations
TL;DR: The results add to the evidence that cryptic species are prevalent in tropical regions, a critical issue in efforts to document global species richness, and illustrate the value of DNA barcoding, especially when coupled with traditional taxonomic tools, in disclosing hidden diversity.
Abstract: Astraptes fulgerator, first described in 1775, is a common and widely distributed neotropical skipper butterfly (Lepidoptera: Hesperiidae). We combine 25 years of natural history observations in northwestern Costa Rica with morphological study and DNA barcoding of museum specimens to show that A. fulgerator is a complex of at least 10 species in this region. Largely sympatric, these taxa have mostly different caterpillar food plants, mostly distinctive caterpillars, and somewhat different ecosystem preferences but only subtly differing adults with no genitalic divergence. Our results add to the evidence that cryptic species are prevalent in tropical regions, a critical issue in efforts to document global species richness. They also illustrate the value of DNA barcoding, especially when coupled with traditional taxonomic tools, in disclosing hidden diversity.
3,112 citations
TL;DR: Five sequences coding for proteins homologous to components of the respiratory‐chain NADH dehydrogenase from human mitochondria have been found and sequence and expression analyses indicate both prokaryotic and eukaryotic features of the chloroplast genes.
Abstract: The complete nucleotide sequence (155 844 bp) of tobacco (Nicotiana tabacum var. Bright Yellow 4) chloroplast DNA has been determined. It contains two copies of an identical 25 339 bp inverted repeat, which are separated by a 86 684 bp and a 18 482 bp single-copy region. The genes for 4 different rRNAs, 30 different tRNAs, 39 different proteins and 11 other predicted protein coding genes have been located. Among them, 15 genes contain introns. Blot hybridization revealed that all rRNA and tRNA genes and 27 protein genes so far analysed are transcribed in the chloroplast and that primary transcripts of the split genes hitherto examined are spliced. Five sequences coding for proteins homologous to components of the respiratory-chain NADH dehydrogenase from human mitochondria have been found. The 30 tRNAs predicted from their genes are sufficient to read all codons if the ;two out of three' and ;U:N wobble' mechanisms operate in the chloroplast. Two sequences which autonomously replicate in yeast have also been mapped. The sequence and expression analyses indicate both prokaryotic and eukaryotic features of the chloroplast genes.
2,184 citations
TL;DR: The finding of large COI sequence differences between, as compared to small differences within, species confirms the effectiveness of COI barcodes for the identification of bird species, and implies that a standard screening threshold of sequence difference could speed the discovery of new animal species.
Abstract: Short DNA sequences from a standardized region of the genome provide a DNA barcode for identifying species. Compiling a public library of DNA barcodes linked to named specimens could provide a new master key for identifying species, one whose power will rise with increased taxon coverage and with faster, cheaper sequencing. Recent work suggests that sequence diversity in a 648-bp region of the mitochondrial gene, cytochrome c oxidase I (COI), might serve as a DNA barcode for the identification of animal species. This study tested the effectiveness of a COI barcode in discriminating bird species, one of the largest and best-studied vertebrate groups. We determined COI barcodes for 260 species of North American birds and found that distinguishing species was generally straightforward. All species had a different COI barcode(s), and the differences between closely related species were, on average, 18 times higher than the differences within species. Our results identified four probable new species of North American birds, suggesting that a global survey will lead to the recognition of many additional bird species. The finding of large COI sequence differences between, as compared to small differences within, species confirms the effectiveness of COI barcodes for the identification of bird species. This result plus those from other groups of animals imply that a standard screening threshold of sequence difference (10× average intraspecific difference) could speed the discovery of new animal species. The growing evidence for the effectiveness of DNA barcodes as a basis for species identification supports an international exercise that has recently begun to assemble a comprehensive library of COI sequences linked to named specimens.
2,115 citations