DOCSLIB.ORG

DNA Barcoding in Molecular Identification and Phylogenetic Relationship of Beneficial Wild

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
DNA Barcoding in Molecular Identification and Phylogenetic Relationship of Beneficial Wild ORIGINAL ARTICLE Bali Medical Journal (Bali MedJ) 2021, Volume 10, Number 1: 82-88 P-ISSN.2089-1180, E-ISSN: 2302-2914 DNA barcoding in molecular identification and phylogenetic relationship of beneficial wild Published by Bali Medical Journal Balinese red algae, Bulung sangu (Gracilaria sp.) I Gede Putu Wirawan1, Maria Malida Vernandes Sasadara1,2*, I Nyoman Wijaya1, Anak Agung Keswari Krinandika1 ABSTRACT Background: Bulung sangu (Gracilaria sp). is wildly widespread Rhodophyta in Bali and usually consumed as vegetable. Bulung sangu is reported for its bioactive compound scientifically proven as antioxidant, anti-inflammatory, and anti- hypercholesterolemia. Likewise, Bulung sangu as Rhodophyta is a potential source for food, fertilizer, cosmetic and 1Faculty of Agriculture, Universitas Udayana, Denpasar, Bali, Indonesia; pharmaceutical industry. Bulung sangu production is highly fluctuating and unable to meet its demands. Proper and correct 2Department of Natural Medicine, cultivation methods based on molecular information are expected to increase the availability of Bulung sangu in Bali. Faculty of Pharmacy, Universitas Complete taxonomy is required to design the proper cultivation method. Meanwhile, Bulung sangu taxonomy is limited to Mahasaraswati, Denpasar, Bali, the genus level. More information on Bulung sangu strain is needed, either does its relatedness. Indonesia Methods: Fresh Bulung sangu was collected from Serangan coastal area, Bali. DNA extraction was applied followed by PCR amplification using six combinations of COI primer sequence. DNA sequences obtained was evaluated to determine pairwise *Corresponding author: distance, percent of similarity and phylogenetic relationship compared to Gracilaria species registered in GenBank. Maria Malida Vernandes Sasadara; Results: PCR amplification produced 730 base pairs amplicons. Genetic distance and percentage of similarity obtained Faculty of Agriculture, Universitas exhibit relatedness to Gracilaria gracilis with 0.487 of pairwise distance and 49.04% of similarity. The phylogenetic tree Udayana, Denpasar, Bali, Indonesia; produced seven clades in which Bulung sangu and Gracilaria gracilis were in the same clade. Department of Natural Medicine, Faculty Conclusion: Bulung sangu showed closest relatedness to Gracilaria gracilis. of Pharmacy, Universitas Mahasaraswati, Denpasar, Bali, Indonesia; [email protected] Keywords: COI gene, Gracilariaceae, Macroalgae, Molecular Identification, Rhodophyta. Cite This Article: Wirawan, I.G.P., Sasadara, M.M.V., Wijaya, I.N., Krinandika, A.A.K. 2021. DNA barcoding in molecular Received: 2020-11-27 identification and phylogenetic relationship of beneficial wild Balinese red algae, Bulung sangu (Gracilaria sp.). Bali Medical Accepted: 2021-03-16 Journal 10(1): 82-88. DOI: 10.15562/bmj.v10i1.2093 Published: 2021-04-01 INTRODUCTION sp. is also potentially developed in the for seaweed. Local production is merely cosmetics and pharmaceutical industries.1 sufficient to supply the local markets and Seaweed or macroalgae has long been The previous study reported several Java trading. Bulung sangu has not been a product widely consumed by people beneficial phytochemicals inBulung cultivated yet in Bali, even though some around the world. Along with science and sangu ethanolic extract, which exhibited coastal areas are felicitous for seaweed technology development, seaweed has strong antioxidant and anti-inflammatory cultivation. Seaweed availability in Bali become very diverse both for food and activity against UV-radiation-induced skin is highly fluctuating. Proper cultivation non-food products. The Gracilariaceae damage.2 Several species of Gracilaria also is expected to maintain the availability has emerged as one of the families with reported various pharmacological activity, of seaweed so that it fulfills the global economic potential. Indonesia has including immunostimulator, allelopathic demands. several varieties of Gracilaria sp. with activity, antibacterial, antifungal, and Genetic information is useful in different morphological and anatomical antiviral, effects for cardiovascular and cultivation management and increases characteristics. Gracilaria sp. is claimed gastrointestinal.3–5 cultivation and conservation success.6 It under various names such as sango- The excessive growth ofBulung sangu provides a sufficient amount, especially sango, rambu kasang, janggut dayung, has been reported in harvest season in to be developed in pharmaceutical dongi-dongi, bulung embulung, agar- many parts of coastal areas in Bali and and cosmetics needs. Genetic study is agar karang, agar-agar jahe, and Bulung will be decreasing in another season. also needed for species that challenge sangu. Bulung sangu is a Gracilaria sp. Nowadays, seaweed production in Bali extinction and limited distribution. that grows wildly in Bali and is commonly is unable to meet Indonesia’s demand Genetic helps in areas mapping for consumed as a vegetable. Gracilaria 82 Published by Bali Medical Journal | Bali Medical Journal 2021; 10(1):Open 82-88 access: | doi: www.balimedicaljournal.org 10.15562/bmj.v10i1.2093 ORIGINAL ARTICLE species with high genetic variability as a conservation and species identification.7 Understanding natural populations’ ecology and genetic structure is essential to determine strategies for conservation, nurseries, and sustainable management.8 Information about Bulung sangu taxonomy is limited to the genus level. High morphological variation in the wild and the lack of morphological diagnostics have misidentified seaweed strains, impacting the wrong cultivation method and declining productivity and quality of cultivated seaweed.1 Strain identification relying on morphological analysis is relatively difficult to establish Figure 1. The sampling sites in Serangan (shows in arrow), -8.719715,115.242034 due to simple morphology and anatomy, convergence, rampant phenotypic plasticity, and alternation of heteromorphic generations.9 Identification based on complete taxonomy helps to correctly and effectively identify the seaweed strains.1 DNA Barcoding provides fast and accurate system for species identification. Instead of using whole-genome, DNA Barcoding uses short DNA sequence generated from the standard region of the genome known as marker. The specific locus used in DNA Barcodes must be determined on most taxa that are sought and sequenced without using specific Polymerase Chain Reaction (PCR) primers.10 Several markers have been developed previously to identify algae, such as Internal Transcribed Spacer (ITS) in the ribosomal cistron, rubisco operon, and variable parts of large ribosomal cistron subunits; nevertheless, those markers serve unsatisfactory shortcoming. The lack of universal markers resulted in multiple, independent, and not easily comparable system. Nonetheless, molecular detection using markers designed from the gene cytochrome oxidase subunit I (COI) was reported generating powerful ally in the identification of red algal species.9 This research was aimed to map the relatedness of Bulung sangu using COI gene as the barcoding marker. Figure 2. Thallus of Bulung sangu (A). Bulung sangu lives by attaching itself to plants or other seaweed using holdfast (arrow), which has a root-like structure in a MATERIAL AND METHODS true plant (B), thus wraps the thallus on other seaweed (C) Study area The study was Test-tube Lab Research, carried out from July 2019 until April Universitas Udayana and Laboratory of Algae material 2020 on Serangan Island (Bali), Central Genetica Science Indonesia. Samples of fresh Bulung sangu were Laboratory of Genetic Resources collected from Serangan coastal area, Bali Published by Bali Medical Journal | Bali Medical Journal 2021; 10(1): 82-88 | doi: 10.15562/bmj.v10i1.2093 83 ORIGINAL ARTICLE primers. PCR amplification was held using Toyobo KOD FX Neo Kit. Amplification reaction followed the kit procedure with one μL of DNA template was used. PCR conditions were used as follows: 1 cycle of 95°C for 2 min, 35 cycles of 95°C for 10s, annealing temperature (Table 2) for 30 sec and 72°C for 1 min. The PCR products (2 μl) were visualized on 1% agarose gel. Primer combination resulting in amplification was then cloned in eight colonies of pTA2 vector plasmid. Data analysis The COI sequence was determined by bidirectional sequencing. The obtained sequence was then analyzed using the BLAST (Basic Local Alignment Search Tool) program in the NCBI (National Center for Biotechnology Information) Figure 3. Electrophoresis of PCR products. Primer optimization generated six database (https://www.ncbi.nlm.nih.gov/). primer combinations. Combination of GWSF-GWSR that was annealed in Ten accession numbers of Gracilaria temperature of 50°C produced clear band. Primer combination of GWSF- species registered in the NCBI database GWSR5 and GWSF5-GWSR5 did not produced amplification. with the highest percent identity to Bulung sangu sequence were used to determine pairwise distance, percent of similarity and construct the phylogenetic tree. Sequences were aligned using Muscle, under default parameter using MEGA program (version 6.0). Genetic distance and percent similarity were then obtained following the phylogenetic tree construction by maximum parsimony. The reliability of the branches was evaluated with non-parametric bootstrapping (1000 replication). Figure 4. Phylogram of Bulung sangu constructed using maximum parsimony RESULTS Bulung sangu was obtained in the intertidal (Figure 1). Bulung sangu samples were optical
Load more

Recommended publications
  • Using DNA Barcodes to Identify and Classify Living Things
    Using DNA Barcodes to Identify and Classify Living Things www.dnabarcoding101.org LABORATORY Using DNA Barcodes to Identify and Classify Living Things OBJECTIVES This laboratory demonstrates several important concepts of modern biology. During this laboratory, you will: • Collect and analyze sequence data from plants, fungi, or animals—or products made from them. • Use DNA sequence to identify species. • Explore relationships between species. In addition, this laboratory utilizes several experimental and bioinformatics methods in modern biological research. You will: • Collect plants, fungi, animals, or products in your local environment or neighborhood. • Extract and purify DNA from tissue or processed material. • Amplify a specific region of the chloroplast, mitochondrial, or nuclear genome by polymerase chain reaction (PCR) and analyze PCR products by gel electrophoresis. • Use the Basic Local Alignment Search Tool (BLAST) to identify sequences in databases. • Use multiple sequence alignment and tree-building tools to analyze phylogenetic relation - ships. INTRODUCTION Taxonomy, the science of classifying living things according to shared features, has always been a part of human society. Carl Linneas formalized biological classifi - cation with his system of binomial nomenclature that assigns each organism a genus and species name. Identifying organisms has grown in importance as we monitor the biological effects of global climate change and attempt to preserve species diversity in the face of accelerating habitat destruction. We know very little about the diversity of plants and animals—let alone microbes—living in many unique ecosystems on earth. Less than two million of the estimated 5–50 million plant and animal species have been identified. Scientists agree that the yearly rate of extinction has increased from about one species per million to 100–1,000 species per million.
    [Show full text]
  • DNA Barcoding Distinguishes Pest Species of the Black Fly Genus <I
    University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln Faculty Publications: Department of Entomology Entomology, Department of 11-2013 DNA Barcoding Distinguishes Pest Species of the Black Fly Genus Cnephia (Diptera: Simuliidae) I. M. Confitti University of Toronto K. P. Pruess University of Nebraska-Lincoln A. Cywinska Ingenomics, Inc. T. O. Powers University of Nebraska-Lincoln D. C. Currie University of Toronto and Royal Ontario Museum, [email protected] Follow this and additional works at: http://digitalcommons.unl.edu/entomologyfacpub Part of the Entomology Commons Confitti, I. M.; Pruess, K. P.; Cywinska, A.; Powers, T. O.; and Currie, D. C., "DNA Barcoding Distinguishes Pest Species of the Black Fly Genus Cnephia (Diptera: Simuliidae)" (2013). Faculty Publications: Department of Entomology. 616. http://digitalcommons.unl.edu/entomologyfacpub/616 This Article is brought to you for free and open access by the Entomology, Department of at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Faculty Publications: Department of Entomology by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. MOLECULAR BIOLOGY/GENOMICS DNA Barcoding Distinguishes Pest Species of the Black Fly Genus Cnephia (Diptera: Simuliidae) 1,2 3 4 5 1,2,6 I. M. CONFLITTI, K. P. PRUESS, A. CYWINSKA, T. O. POWERS, AND D. C. CURRIE J. Med. Entomol. 50(6): 1250Ð1260 (2013); DOI: http://dx.doi.org/10.1603/ME13063 ABSTRACT Accurate species identiÞcation is essential for cost-effective pest control strategies. We tested the utility of COI barcodes for identifying members of the black ßy genus Cnephia Enderlein (Diptera: Simuliidae). Our efforts focus on four Nearctic Cnephia speciesÑCnephia dacotensis (Dyar & Shannon), Cnephia eremities Shewell, Cnephia ornithophilia (Davies, Peterson & Wood), and Cnephia pecuarum (Riley)Ñthe latter two being current or potential targets of biological control programs.
    [Show full text]
  • DNA Barcoding Analysis and Phylogenetic Relation of Mangroves in Guangdong Province, China
    Article DNA Barcoding Analysis and Phylogenetic Relation of Mangroves in Guangdong Province, China Feng Wu 1,2 , Mei Li 1, Baowen Liao 1,*, Xin Shi 1 and Yong Xu 3,4 1 Key Laboratory of State Forestry Administration on Tropical Forestry Research, Research Institute of Tropical Forestry, Chinese Academy of Forestry, Guangzhou 510520, China; [email protected] (F.W.); [email protected] (M.L.); [email protected] (X.S.) 2 Zhaoqing Xinghu National Wetland Park Management Center, Zhaoqing 526060, China 3 Key Laboratory of Plant Resources Conservation and Sustainable Utilization, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, China; [email protected] 4 University of Chinese Academy of Sciences, Beijing 100049, China * Correspondence: [email protected]; Tel.: +86-020-8702-8494 Received: 9 December 2018; Accepted: 4 January 2019; Published: 12 January 2019 Abstract: Mangroves are distributed in the transition zone between sea and land, mostly in tropical and subtropical areas. They provide important ecosystem services and are therefore economically valuable. DNA barcoding is a useful tool for species identification and phylogenetic reconstruction. To evaluate the effectiveness of DNA barcoding in identifying mangrove species, we sampled 135 individuals representing 23 species, 22 genera, and 17 families from Zhanjiang, Shenzhen, Huizhou, and Shantou in the Guangdong province, China. We tested the universality of four DNA barcodes, namely rbcL, matK, trnH-psbA, and the internal transcribed spacer of nuclear ribosomal DNA (ITS), and examined their efficacy for species identification and the phylogenetic reconstruction of mangroves. The success rates for PCR amplification of rbcL, matK, trnH-psbA, and ITS were 100%, 80.29% ± 8.48%, 99.38% ± 1.25%, and 97.18% ± 3.25%, respectively, and the rates of DNA sequencing were 100%, 75.04% ± 6.26%, 94.57% ± 5.06%, and 83.35% ± 4.05%, respectively.
    [Show full text]
  • Cristescu TREE 2014.Pdf
    TREE-1853; No. of Pages 6 Opinion From barcoding single individuals to metabarcoding biological communities: towards an integrative approach to the study of global biodiversity Melania E. Cristescu Department of Biology, McGill University, Montreal, QC H3A 1B1, Canada DNA-based species identification, known as barcoding, introduced by Arnot et al. [6] and was firmly advanced transformed the traditional approach to the study of and standardized by Hebert et al. [7]. The simple idea of biodiversity science. The field is transitioning from bar- using a short DNA fragment as a barcode (see Glossary) for coding individuals to metabarcoding communities. This identifying species across the Metazoa has been both revolution involves new sequencing technologies, bio- strongly embraced and vigorously scrutinized over the past informatics pipelines, computational infrastructure, and decade [8,9]. Nevertheless, the efforts led by Paul Hebert, experimental designs. In this dynamic genomics land- and supported by the Consortium for the Barcode of Life scape, metabarcoding studies remain insular and biodi- (CBoL; http://www.barcodeoflife.org/) resulted in a global versity estimates depend on the particular methods enterprise that combined molecular tools with valuable but used. In this opinion article, I discuss the need for a scarce taxonomic expertise [10,11]. Today, DNA barcodes coordinated advancement of DNA-based species identi- are being used commonly to identify specimens and the fication that integrates taxonomic and barcoding infor- approach has wide applications in biodiversity conserva- mation. Such an approach would facilitate access to tion, environmental management, invasion biology, the almost 3 centuries of taxonomic knowledge and 1 de- study of trophic interactions, and food safety [12–14].
    [Show full text]
  • Automated, Phylogeny-Based Genotype Delimitation of the Hepatitis Viruses HBV and HCV
    Automated, phylogeny-based genotype delimitation of the Hepatitis Viruses HBV and HCV Dora Serdari1, Evangelia-Georgia Kostaki2, Dimitrios Paraskevis2, Alexandros Stamatakis1,3 and Paschalia Kapli4 1 The Exelixis Lab, Scientific Computing Group, Heidelberg Institute for Theoretical Studies, Heidelberg, Germany 2 Department of Hygiene Epidemiology and Medical Statistics, School of Medicine, National and Kapodistrian University of Athens, Athens, Greece 3 Institute for Theoretical Informatics, Karlsruhe Institute of Technology, Karlsruhe, Germany 4 Centre for Life's Origins and Evolution, Department of Genetics Evolution and Environment, University College London, University of London, London, United Kingdom ABSTRACT Background. The classification of hepatitis viruses still predominantly relies on ad hoc criteria, i.e., phenotypic traits and arbitrary genetic distance thresholds. Given the subjectivity of such practices coupled with the constant sequencing of samples and discovery of new strains, this manual approach to virus classification becomes cumbersome and impossible to generalize. Methods. Using two well-studied hepatitis virus datasets, HBV and HCV, we assess if computational methods for molecular species delimitation that are typically applied to barcoding biodiversity studies can also be successfully deployed for hepatitis virus classification. For comparison, we also used ABGD, a tool that in contrast to other distance methods attempts to automatically identify the barcoding gap using pairwise genetic distances for a set of aligned input sequences. Results—Discussion. We found that the mPTP species delimitation tool identified even without adapting its default parameters taxonomic clusters that either correspond to the currently acknowledged genotypes or to known subdivision of genotypes Submitted 17 April 2019 (subtypes or subgenotypes). In the cases where the delimited cluster corresponded Accepted 26 August 2019 Published 25 October 2019 to subtype or subgenotype, there were previous concerns that their status may be underestimated.
    [Show full text]
  • Dna Barcodes, Partitioned Phylogenetic Models, And
    LARGE DATASETS AND TRICHOPTERA PHYLOGENETICS: DNA BARCODES, PARTITIONED PHYLOGENETIC MODELS, AND THE EVOLUTION OF PHRYGANEIDAE By PAUL BRYAN FRANDSEN A dissertation submitted to the Graduate School-New Brunswick Rutgers, The State University of New Jersey In partial fulfillment of the requirements For the degree of Doctor of Philosophy Graduate Program in Entomology Written under the direction of Karl M. Kjer And approved by _____________________________________ _____________________________________ _____________________________________ _____________________________________ New Brunswick, New Jersey OCTOBER 2015 ABSTRACT OF THE DISSERTATION Large datasets and Trichoptera phylogenetics: DNA barcodes, partitioned phylogenetic models, and the evolution of Phryganeidae By PAUL BRYAN FRANDSEN Dissertation Director: Karl M. Kjer Large datasets in phylogenetics—those with a large number of taxa, e.g. DNA barcode data sets, and those with a large amount of sequence data per taxon, e.g. data sets generated from high throughput sequencing—pose both exciting possibilities and interesting analytical problems. The analysis of both types of large datasets is explored in this dissertation. First, the use of DNA barcodes in phylogenetics is investigated via the generation of phylogenetic trees for known monophyletic clades. Barcodes are found to be useful in shallow scale phylogenetic analyses when given a well-supported scaffold on which to place them. One of the analytical challenges posed by large phylogenetic datasets is the selection of appropriate partitioned models of molecular evolution. The most commonly used model partitioning strategies can fail to characterize the true variation of the evolutionary process and this effect can be exacerbated when applied to large datasets. A new, scalable algorithm for the automatic selection ! ii! of partitioned models of molecular evolution is proposed with an eye toward reducing systematic error in phylogenomics.
    [Show full text]
  • DNA Barcoding: How It Complements Taxonomy, Molecular Phylogenetics and Population Genetics
    Opinion TRENDS in Genetics Vol.23 No.4 DNA barcoding: how it complements taxonomy, molecular phylogenetics and population genetics Mehrdad Hajibabaei1, Gregory A.C. Singer2, Paul D.N. Hebert1 and Donal A. Hickey3 1 Biodiversity Institute of Ontario, Department of Integrative Biology, University of Guelph, Guelph, Ontario N1G 2W1, Canada 2 Human Cancer Genetics Program, The Ohio State University, Columbus, OH 43210, USA 3 Department of Biology, Concordia University, 7141 Sherbrooke Street, Montreal, Quebec H4B 1R6, Canada DNA barcoding aims to provide an efficient method for barcoding datasets are essentially composed of short species-level identifications and, as such, will contribute DNA sequences from several individuals of a large number powerfully to taxonomic and biodiversity research. As of species (typically five to ten individuals per species, but the number of DNA barcode sequences accumulates, these numbers will increase in the future) (Figure 1). however, these data will also provide a unique ‘horizon- Here, we discuss the role of DNA barcodes in advancing tal’ genomics perspective with broad implications. For the taxonomic enterprise and its potential to provide a example, here we compare the goals and methods of contextual framework for both building phylogenies and DNA barcoding with those of molecular phylogenetics for population genetics. In particular, we argue that bar- and population genetics, and suggest that DNA barcod- code results can be of high value in aiding the selection of ing can complement current research in these areas by species for more detailed analysis, and demonstrate that providing background information that will be helpful in DNA barcoding can broaden our understanding of both the selection of taxa for further analyses.
    [Show full text]
  • DNA Barcoding Reveal Patterns of Species Diversity Among
    www.nature.com/scientificreports OPEN DNA barcoding reveal patterns of species diversity among northwestern Pacific molluscs Received: 04 April 2016 Shao’e Sun, Qi Li, Lingfeng Kong, Hong Yu, Xiaodong Zheng, Ruihai Yu, Lina Dai, Yan Sun, Accepted: 25 August 2016 Jun Chen, Jun Liu, Lehai Ni, Yanwei Feng, Zhenzhen Yu, Shanmei Zou & Jiping Lin Published: 19 September 2016 This study represents the first comprehensive molecular assessment of northwestern Pacific molluscs. In total, 2801 DNA barcodes belonging to 569 species from China, Japan and Korea were analyzed. An overlap between intra- and interspecific genetic distances was present in 71 species. We tested the efficacy of this library by simulating a sequence-based specimen identification scenario using Best Match (BM), Best Close Match (BCM) and All Species Barcode (ASB) criteria with three threshold values. BM approach returned 89.15% true identifications (95.27% when excluding singletons). The highest success rate of congruent identifications was obtained with BCM at 0.053 threshold. The analysis of our barcode library together with public data resulted in 582 Barcode Index Numbers (BINs), 72.2% of which was found to be concordantly with morphology-based identifications. The discrepancies were divided in two groups: sequences from different species clustered in a single BIN and conspecific sequences divided in one more BINs. In Neighbour-Joining phenogram, 2,320 (83.0%) queries fromed 355 (62.4%) species-specific barcode clusters allowing their successful identification. 33 species showed paraphyletic and haplotype sharing. 62 cases are represented by deeply diverged lineages. This study suggest an increased species diversity in this region, highlighting taxonomic revision and conservation strategy for the cryptic complexes.
    [Show full text]
  • DNA Barcoding Central Asian Butterflies: Increasing Geographical
    Molecular Ecology Resources (2009) 9, 1302–1310 doi: 10.1111/j.1755-0998.2009.02577.x DNA BARCODING DNA barcoding Central Asian butterflies: increasing geographical dimension does not significantly reduce the success of species identification VLADIMIR A. LUKHTANOV,*† ANDREI SOURAKOV,‡ EVGENY V. ZAKHAROV§ and PAUL D. N. HEBERT§ *Department of Karyosystematics, Zoological Institute of Russian Academy of Science, Universitetskaya nab. 1, 199034 St. Petersburg, Russia, †Department of Entomology, St. Petersburg State University, Universitetskaya nab. 7/9, 199034 St. Petersburg, Russia, ‡McGuire Center for Lepidoptera and Biodiversity, Florida Museum of Natural History, University of Florida, Gainesville, FL 32611, USA, §Biodiversity Institute of Ontario, University of Guelph, Guelph, ON, Canada N1G 2W1 Abstract DNA barcoding employs short, standardized gene regions (5’ segment of mitochondrial cytochrome oxidase subunit I for animals) as an internal tag to enable species identification. Prior studies have indicated that it performs this task well, because interspecific variation at cytochrome oxidase subunit I is typically much greater than intraspecific variation. How- ever, most previous studies have focused on local faunas only, and critics have suggested two reasons why barcoding should be less effective in species identification when the geograph- ical coverage is expanded. They suggested that many recently diverged taxa will be excluded from local analyses because they are allopatric. Second, intraspecific variation may be seri- ously underestimated by local studies, because geographical variation in the barcode region is not considered. In this paper, we analyse how adding a geographical dimension affects barcode resolution, examining 353 butterfly species from Central Asia. Despite predictions, we found that geographically separated and recently diverged allopatric species did not show, on average, less sequence differentiation than recently diverged sympatric taxa.
    [Show full text]
  • Teleostei: Beryciformes: Holocentridae): Reconciling More Than 100 Years of Taxonomic Confusion ⇑ Alex Dornburg A, , Jon A
    Molecular Phylogenetics and Evolution 65 (2012) 727–738 Contents lists available at SciVerse ScienceDirect Molecular Phylogenetics and Evolution journal homepage: www.elsevier.com/locate/ympev Molecular phylogenetics of squirrelfishes and soldierfishes (Teleostei: Beryciformes: Holocentridae): Reconciling more than 100 years of taxonomic confusion ⇑ Alex Dornburg a, , Jon A. Moore b,c, Rachel Webster a, Dan L. Warren d, Matthew C. Brandley e, Teresa L. Iglesias f, Peter C. Wainwright g, Thomas J. Near a,h a Department of Ecology and Evolutionary Biology, Yale University, New Haven, CT 06520, USA b Florida Atlantic University, Wilkes Honors College, Jupiter, FL 33458, USA c Florida Atlantic University, Harbor Branch Oceanographic Institution, Fort Pierce, FL 34946, USA d Section of Integrative Biology, University of Texas, Austin, TX 78712, USA e School of Biological Sciences, University of Sydney, NSW 2006, Australia f Graduate Group in Animal Behavior, University of California, Davis, CA 95616, USA g Department of Evolution and Ecology, University of California, Davis, CA 95616, USA h Peabody Museum of Natural History, Yale University, New Haven, CT 06520, USA article info abstract Article history: Squirrelfishes and soldierfishes (Holocentridae) are among the most conspicuous species in the nocturnal Received 16 April 2012 reef fish community. However, there is no clear consensus regarding their evolutionary relationships, Revised 19 July 2012 which is reflected in a complicated taxonomic history. We collected DNA sequence data from multiple Accepted 23 July 2012 single copy nuclear genes and one mitochondrial gene sampled from over fifty percent of the recognized Available online 3 August 2012 holocentrid species and infer the first species-level phylogeny of the Holocentridae.
    [Show full text]
  • Assessing the Phylogenetic Utility of DNA Barcoding Using the New Zealand Cicada Genus Kikihia Megan Ribak University of Connecticut - Storrs, [email protected]
    University of Connecticut OpenCommons@UConn Honors Scholar Theses Honors Scholar Program Spring 5-9-2010 Assessing the Phylogenetic Utility of DNA Barcoding Using the New Zealand Cicada Genus Kikihia Megan Ribak University of Connecticut - Storrs, [email protected] Follow this and additional works at: https://opencommons.uconn.edu/srhonors_theses Part of the Biology Commons, Developmental Biology Commons, and the Genetics Commons Recommended Citation Ribak, Megan, "Assessing the Phylogenetic Utility of DNA Barcoding Using the New Zealand Cicada Genus Kikihia" (2010). Honors Scholar Theses. 127. https://opencommons.uconn.edu/srhonors_theses/127 1 APPROVAL PAGE HONORS THESIS ASSESSING THE PHYLOGENETIC UTILITY OF DNA BARCODING USING THE NEW ZEALAND CICADA GENUS KIKIHIA Presented by Megan Pierce Ribak Honors Thesis Advisor_____________________________________________________ Chris Simon Honors Academic Advisor__________________________________________________ Janine Caira Associate Honors Advisor__________________________________________________ Elizabeth Jockusch Department of Ecology & Evolutionary Biology The University of Connecticut 2010 2 Abstract DNA Barcoding (Hebert et al. 2003) has the potential to revolutionize the process of identifying and cataloguing biodiversity; however, significant controversy surrounds some of the proposed applications. In the seven years since DNA barcoding was introduced, the Web of Science records more than 600 studies that have weighed the pros and cons of this procedure. Unfortunately, the scientific community has been unable to come to any consensus on what threshold to use to differentiate species or even whether the barcoding region provides enough information to serve as an accurate species identification tool. The purpose of my thesis is to analyze mitochondrial DNA (mtDNA) barcoding’s potential to identify known species and provide a well-resolved phylogeny for the New Zealand cicada genus Kikihia .
    [Show full text]
  • DNA Barcoding and Its Applications
    International Journal of Engineering Research and General Science Volume 3, Issue 2, Part 2, March-April, 2015 ISSN 2091-2730 DNA Barcoding and Its Applications Sukhamrit Kaur [email protected] Contact No.: 09872455529 Abstract— DNA barcoding is a system for fast and accurate species identification that makes ecological system more accessible by using short DNA sequence instead of whole genome and is used for eukaryotes. The short DNA sequence is generated from standard region of genome known as marker. This marker is different for various species like CO1 cytochrome c oxidase 1 for animals, matK for plants and Internal Transcribed Spacer (ITS) for fungus. DNA barcoding has many applications in various fields like preserving natural resources, protecting endangered species, controlling agriculture pests, identifying disease vectors, monitoring water quality, authentication of natural health products and identification of medicinal plants. Keywords— CO1, DNA, DNA Barcode, IBOL, Identification, ITS, Marker. INTRODUCTION Biological effects of global climate lead to importance of identification of organisms to preserve species because of increasing habitat destruction. About 5 to 50 million plants and animals are living on earth, out of which less than 2 million have been identified. Extinction of animals and plants is increasing yearly means thousand of them are lost each year and most of them are not identified yet.[1] This destruction and endangerment of ecosystem has lead to an improved system for identifying species. In recent years new ecological approach called DNA barcoding has been proposed to identify species and ecology research. [2][3] DNA barcoding, a system for fast and accurate species identification which will make ecological system more accessible.
    [Show full text]