DNA Barcode Authentication of Wood Samples of Threatened and Commercial Timber Trees within the Tropical Dry Evergreen Forest of India

Background India is rich with biodiversity, which includes a large number of endemic, rare and threatened plant species. Previous studies have used DNA barcoding to inventory species for applications in biodiversity monitoring, conservation impact assessment, monitoring of illegal trading, authentication of traded medicinal plants etc. This is the first tropical dry evergreen forest (TDEF) barcode study in the World and the first attempt to assemble a reference barcode library for the trees of India as part of a larger project initiated by this research group. Methodology/Principal Findings We sampled 429 trees representing 143 tropical dry evergreen forest (TDEF) species, which included 16 threatened species. DNA barcoding was completed using rbcL and matK markers. The tiered approach (1st tier rbcL; 2nd tier matK) correctly identified 136 out of 143 species (95%). This high level of species resolution was largely due to the fact that the tree species were taxonomically diverse in the TDEF. Ability to resolve taxonomically diverse tree species of TDEF was comparable among the best match method, the phylogenetic method, and the characteristic attribute organization system method. Conclusions We demonstrated the utility of the TDEF reference barcode library to authenticate wood samples from timber operations in the TDEF. This pilot research study will enable more comprehensive surveys of the illegal timber trade of threatened species in the TDEF. This TDEF reference barcode library also contains trees that have medicinal properties, which could be used to monitor unsustainable and indiscriminate collection of plants from the wild for their medicinal value.

[1]  J. Harborne,et al.  Molecular Phylogeny of Fabaceae based on rbcL sequence data: With special emphasis on the tribe Mimoseae (Mimosoideae) , 2003 .

[2]  Peter H. Raven,et al.  Advances in legume systematics , 1981 .

[3]  A. Fazekas,et al.  Testing candidate plant barcode regions in the Myristicaceae , 2008, Molecular ecology resources.

[4]  R. Jorgensen,et al.  Ribosomal DNA spacer-length polymorphisms in barley: mendelian inheritance, chromosomal location, and population dynamics. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[5]  V. Grant IN PLANT SPECIES , 1966 .

[6]  Steven G Newmaster,et al.  DNA barcoding detects contamination and substitution in North American herbal products , 2013, BMC Medicine.

[7]  Georgina M Mace,et al.  The role of taxonomy in species conservation. , 2004, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[8]  N. Parthasarathy,et al.  Tropical Dry Evergreen Forests of Peninsular India: Ecology and Conservation Significance , 2008 .

[9]  Gaurav Vaidya,et al.  DNA barcoding and taxonomy in Diptera: a tale of high intraspecific variability and low identification success. , 2006, Systematic biology.

[10]  D. Stevenson,et al.  Character-based, population-level DNA barcoding in Mexican species of Zamia L. (Zamiaceae: Cycadales) , 2010, Mitochondrial DNA.

[11]  S. Newmaster,et al.  Molecular taxonomic tools provide more accurate estimates of species richness at less cost than traditional morphology-based taxonomic practices in a vegetation survey , 2014, Biodiversity and Conservation.

[12]  David L. Erickson,et al.  Advances in the Use of DNA Barcodes to Build a Community Phylogeny for Tropical Trees in a Puerto Rican Forest Dynamics Plot , 2010, PloS one.

[13]  Gareth Nelson,et al.  Revisiting the Taxonomic Impediment , 2005, Science.

[14]  Shilin Chen,et al.  Identification of Fabaceae Plants Using the DNA Barcode matK , 2010, Planta medica.

[15]  A. Lambert,et al.  ABGD, Automatic Barcode Gap Discovery for primary species delimitation , 2012, Molecular ecology.

[16]  S. Graham,et al.  Multiple Multilocus DNA Barcodes from the Plastid Genome Discriminate Plant Species Equally Well , 2008, PloS one.

[17]  A. Fazekas,et al.  DNA barcoding in land plants: evaluation of rbcL in a multigene tiered approach , 2006 .

[18]  D. Janzen,et al.  Use of DNA barcodes to identify flowering plants. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[19]  M. Wink,et al.  Molecular Phylogeny of the Papilionoideae (Family Leguminosae): RbcL Gene Sequences versus Chemical Taxonomy , 1995 .

[20]  S. Mathews,et al.  Monophyletic subgroups of the tribe Millettieae (Leguminosae) as revealed by phytochrome nucleotide sequence data , 1998 .

[21]  C. Cruaud,et al.  How Effective Are DNA Barcodes in the Identification of African Rainforest Trees? , 2013, PloS one.

[22]  M. Nei,et al.  MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. , 2011, Molecular biology and evolution.

[23]  J. Doyle,et al.  A phylogeny of the chloroplast gene rbcL in the Leguminosae: taxonomic correlations and insights into the evolution of nodulation. , 1997, American journal of botany.

[24]  Christopher Baraloto,et al.  Identification of Amazonian Trees with DNA Barcodes , 2009, PloS one.

[25]  D. Levin The Nature of Plant Species , 1979, Nature.

[26]  R. J. Bayer,et al.  Molecular phylogenetics of Acacia (Fabaceae: Mimosoideae) based on the chloroplast MATK coding sequence and flanking TRNK intron spacer regions. , 2001, American journal of botany.

[27]  A. Hemerly,et al.  DNA Barcoding Bromeliaceae: Achievements and Pitfalls , 2012, PloS one.

[28]  C. Margules,et al.  Biological Models for Monitoring Species Decline: The Construction and Use of Data Bases , 1994 .

[29]  Nicolas Salamin,et al.  Land plants and DNA barcodes: short-term and long-term goals , 2005, Philosophical Transactions of the Royal Society B: Biological Sciences.

[30]  S. Graham,et al.  Are plant species inherently harder to discriminate than animal species using DNA barcoding markers? , 2009, Molecular ecology resources.

[31]  Bernd Schierwater,et al.  Character-based DNA barcoding: a superior tool for species classification. , 2009, Berliner und Munchener tierarztliche Wochenschrift.

[32]  N. Baeshen,et al.  Biological Identifications Through DNA Barcodes , 2012 .

[33]  Sylvie Duthoit,et al.  DNA barcoding the floras of biodiversity hotspots , 2008, Proceedings of the National Academy of Sciences.

[34]  R DeSalle,et al.  Character-based DNA barcoding allows discrimination of genera, species and populations in Odonata , 2007, Proceedings of the Royal Society B: Biological Sciences.

[35]  I. Sarkar,et al.  Genetic variation and phylogeny of the cosmopolitan marine genus Tubificoides (Annelida: Clitellata: Naididae: Tubificinae). , 2010, Molecular phylogenetics and evolution.

[36]  Anoop Kumar,et al.  The Internal Transcribed Spacer (ITS) Region and trnhH-psbA Are Suitable Candidate Loci for DNA Barcoding of Tropical Tree Species of India , 2013, PloS one.

[37]  D. Janzen,et al.  Ten species in one: DNA barcoding reveals cryptic species in the neotropical skipper butterfly Astraptes fulgerator. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[38]  J. Ferguson On the use of genetic divergence for identifying species , 2002 .

[39]  P. Hebert,et al.  Identification of the vascular plants of Churchill, Manitoba, using a DNA barcode library , 2012, BMC Ecology.

[40]  Michael G. Simpson,et al.  1 – Plant Systematics: An Overview , 2010 .

[41]  Indra Neil Sarkar,et al.  caos software for use in character‐based DNA barcoding , 2008, Molecular ecology resources.

[42]  De‐Zhu Li,et al.  Comparative analysis of a large dataset indicates that internal transcribed spacer (ITS) should be incorporated into the core barcode for seed plants , 2011, Proceedings of the National Academy of Sciences.

[43]  Jingyuan Song,et al.  Expedient identification of Magnoliaceae species by DNA barcoding , 2014 .

[44]  S. Graham,et al.  Discriminating plant species in a local temperate flora using the rbcL+matK DNA barcode , 2011 .

[45]  X. Ge,et al.  Testing DNA barcoding in closely related groups of Lysimachia L. (Myrsinaceae) , 2012, Molecular ecology resources.

[46]  S. G. Champion,et al.  A revised survey of the forest types of India. , 1968 .

[47]  Rob DeSalle,et al.  The unholy trinity: taxonomy, species delimitation and DNA barcoding , 2005, Philosophical Transactions of the Royal Society B: Biological Sciences.

[48]  Gurcharan Singh,et al.  Plant Systematics , 2016 .

[49]  J. Pettengill,et al.  An evaluation of candidate plant DNA barcodes and assignment methods in diagnosing 29 species in the genus Agalinis (Orobanchaceae). , 2010, American journal of botany.

[50]  Rolando Perez,et al.  Plant DNA barcodes and a community phylogeny of a tropical forest dynamics plot in Panama , 2009, Proceedings of the National Academy of Sciences.

[51]  D. Maddison,et al.  Mesquite: a modular system for evolutionary analysis. Version 2.6 , 2009 .

[52]  L. Consaul,et al.  DNA Barcoding the Canadian Arctic Flora: Core Plastid Barcodes (rbcL + matK) for 490 Vascular Plant Species , 2013, PloS one.

[53]  L. Gielly,et al.  Phylogenetic relationships in the genus Leonardoxa (Leguminosae: Caesalpinioideae) inferred from chloroplast trnL intron and trnL-trnF intergenic spacer sequences. , 2001, American journal of botany.

[54]  S. Schuster Next-generation sequencing transforms today's biology , 2008, Nature Methods.

[55]  Royce Steeves,et al.  Improving sequencing quality from PCR products containing long mononucleotide repeats. , 2010, BioTechniques.

[56]  C. Cruaud,et al.  Deciduous Trees and the Application of Universal DNA Barcodes: A Case Study on the Circumpolar Fraxinus , 2012, PloS one.

[57]  P. GeevanC National Environment Policy , 2004 .

[58]  David L. Erickson,et al.  Exploring Tree-Habitat Associations in a Chinese Subtropical Forest Plot Using a Molecular Phylogeny Generated from DNA Barcode Loci , 2011, PloS one.

[59]  Pradhyumna K. Singh,et al.  Universal Plant DNA Barcode Loci May Not Work in Complex Groups: A Case Study with Indian Berberis Species , 2010, PloS one.

[60]  F. Forest,et al.  Phylogenetic Relationships in the Caesalpinioideae (Leguminosae) as Inferred from Chloroplast trnL Intron Sequences , 2009 .

[61]  Jun-bo Yang,et al.  DNA Barcoding of Nyssaceae (Cornales) and Taxonomic Issues , 2012 .

[62]  G. Amato,et al.  The Real maccoyii: Identifying Tuna Sushi with DNA Barcodes – Contrasting Characteristic Attributes and Genetic Distances , 2009, PloS one.

[63]  N. Parthasarathy,et al.  TROPICAL DRY EVERGREEN FORESTS ON THE COROMANDEL COAST OF INDIA: STRUCTURE, COMPOSITION AND HUMAN DISTURBANCE , 2003 .

[64]  Hilla Peretz,et al.  Ju n 20 03 Schrödinger ’ s Cat : The rules of engagement , 2003 .

[65]  D. Stevenson,et al.  A comparison of algorithms for the identification of specimens using DNA barcodes: examples from gymnosperms , 2007, Cladistics : the international journal of the Willi Hennig Society.

[66]  M. Harrington,et al.  Plant DNA Barcodes Can Accurately Estimate Species Richness in Poorly Known Floras , 2011, PloS one.

[67]  Robert C. Edgar,et al.  MUSCLE: multiple sequence alignment with high accuracy and high throughput. , 2004, Nucleic acids research.

[68]  E. Pothos,et al.  Feeding Induced by Cannabinoids Is Mediated Independently of the Melanocortin System , 2008, PloS one.

[69]  A. Galimberti,et al.  A multi-marker DNA barcoding approach to save time and resources in vegetation surveys , 2012 .

[70]  S. Newmaster,et al.  A tiered barcode authentication tool to differentiate medicinal Cassia species in India. , 2014, Genetics and molecular research : GMR.

[71]  J. B. Simmons,et al.  Convention on International Trade in Endangered Species of Wild Fauna and Flora , 1976 .

[72]  W. John Kress,et al.  A DNA barcode for land plants , 2009, Proceedings of the National Academy of Sciences.

[73]  Damon P. Little,et al.  Choosing and Using a Plant DNA Barcode , 2011, PloS one.