A comparative study of different DNA barcoding markers for the identification of some members of Lamiacaea

Abstract The objective of the present work is to evaluate the efficacy of a DNA barcoding approach as a tool for the recognition of commercial kitchen spices belonging to the Lamiaceae family that are usually sold as enhancers of food flavor. A total of 64 spices samples, encompassing six different genera (i.e. Mentha, Ocimum, Origanum, Salvia, Thymus and Rosmarinus) were processed with a classical DNA barcoding approach by amplifying and sequencing four candidate barcode regions (rpoB, rbcL, matK and trnH-psbA) with universal primers. Results suggest that the non-coding trnH-psbA intergenic spacer is the most suitable marker for molecular spices identification followed by matK, with interspecific genetic distance values ranging between about 0% to 7% and 0% to 5%, respectively. Both markers were almost invariably able to distinguish spices species from closest taxa with the exclusion of samples belonging to the genus Oregano. Moreover, in a context of food traceability the two markers are useful to identify commercial processed spice species (sold as dried plant material). We also evaluated the potential benefits of a multilocus barcode approach over a single-marker and although the most suitable combination was the matK + trhH-psbA, the observed genetic distances values were very similar to the discriminatory performance of the trnH-psbA. Finally, this preliminary work provide clear evidences that the efficacy of a DNA barcoding approach to the recognition of commercial spices is biased by the occurrence of taxonomic criticisms as well as traces of hybridization events within the family Lamiaceae. For this reason, to better define a more practical and standardized DNA barcoding tool for spices traceability, the building of a dedicated aromatic plants database in which all species and cultivars are described (both morphologically and molecularly) is strongly required.

[1]  B. Ledda,et al.  Morphological characterization, essential oil composition and DNA genotyping of Ocimum basilicum L. cultivars , 2004 .

[2]  M. Nei,et al.  Molecular Evolutionary Genetics Analysis , 2007 .

[3]  P. Galli,et al.  DNA barcoding reveals fraudulent substitutions in shark seafood products: The Italian case of “palombo” (Mustelus spp.) , 2010 .

[4]  J. Fletcher,et al.  Composition of essential oils , 1998 .

[5]  M. Gouy,et al.  Relationship between morphological taxonomy and molecular divergence within Crustacea: proposal of a molecular threshold to help species delimitation. , 2006, Molecular phylogenetics and evolution.

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

[7]  Mark W. Chase,et al.  A proposal for a standardised protocol to barcode all land plants , 2007 .

[8]  H. Trindade Molecular biology of aromatic plants and spices. A review. , 2010 .

[9]  Joana Costa,et al.  Monitoring genetically modified soybean along the industrial soybean oil extraction and refining processes by polymerase chain reaction techniques , 2010 .

[10]  J. G. Segarra‐Moragues,et al.  Isolation and characterisation of di and tri nucleotide microsatellite loci in Rosmarinusofficinalis (Lamiaceae), using enriched genomic libraries , 2009, Conservation Genetics.

[11]  G. Drakakaki,et al.  Comparison of essential oils and genetic relationship of Origanum × intercedens to its parental taxa in the island of Crete , 2002 .

[12]  Ting Gao,et al.  Validation of the ITS2 Region as a Novel DNA Barcode for Identifying Medicinal Plant Species , 2010, PloS one.

[13]  J. Starr,et al.  A regional approach to plant DNA barcoding provides high species resolution of sedges (Carex and Kobresia, Cyperaceae) in the Canadian Arctic Archipelago , 2010, Molecular ecology resources.

[14]  P. Hollingsworth,et al.  Stopping the stutter: Improvements in sequence quality from regions with mononucleotide repeats can increase the usefulness of non-coding regions for DNA barcoding , 2010 .

[15]  K. Dhanya,et al.  Molecular marker based adulteration detection in traded food and agricultural commodities of plant origin with special reference to spices. , 2010 .

[16]  Ragupathy Subramanyam,et al.  Testing plant barcoding in a sister species complex of pantropical Acacia (Mimosoideae, Fabaceae) , 2009, Molecular ecology resources.

[17]  R. Harley,et al.  OCIMUM: AN OVERVIEW OF CLASSIFICATION AND RELATIONSHIPS , 1999 .

[18]  M. Chase,et al.  Molecular phylogenetics of Caryophyllales based on nuclear 18S rDNA and plastid rbcL, atpB, and matK DNA sequences. , 2002, American journal of botany.

[19]  Andrea Galimberti,et al.  Identification of poisonous plants by DNA barcoding approach , 2010, International Journal of Legal Medicine.

[20]  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.

[21]  K. Başer,et al.  Composition of the essential oils of Turkish Origanum species with commercial importance , 1993 .

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

[23]  D. Curry A REGIONAL APPROACH , 1974 .

[24]  B. Sasikumar,et al.  PCR Based Detection of Adulteration in the Market Samples of Turmeric Powder , 2004 .

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

[26]  R. Hiltunen,et al.  Basil - The Genus Ocimum , 1999 .

[27]  Peter M Hollingsworth,et al.  Selecting barcoding loci for plants: evaluation of seven candidate loci with species‐level sampling in three divergent groups of land plants , 2009, Molecular ecology resources.

[28]  R. Bruni,et al.  RAPD-based method for the quality control of Mediterranean oregano and its contribution to pharmacognostic techniques. , 2009, Journal of agricultural and food chemistry.

[29]  R. Clark,et al.  Salvia divinorum: exposures reported to a statewide poison control system over 10 years. , 2011, The Journal of emergency medicine.

[30]  W. Friedt,et al.  Intraspecific diversity and relationship between subspecies of Origanum vulgare revealed by comparative AFLP and SAMPL marker analysis , 2009, Plant Systematics and Evolution.

[31]  J. Degenhardt,et al.  Identification and characterization of simple sequence repeat markers from a glandular Origanum vulgare expressed sequence tag , 2008, Molecular ecology resources.

[32]  Andrea Galimberti,et al.  DNA barcoding: a six-question tour to improve users' awareness about the method , 2010, Briefings Bioinform..

[33]  M. Nei,et al.  MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. , 2007, Molecular biology and evolution.

[34]  P. Taberlet,et al.  Hybridization in the section Mentha (Lamiaceae) inferred from AFLP markers. , 2002, American journal of botany.

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

[36]  Unoda The regional approach , 1977 .

[37]  B. Bohanec,et al.  Genetic relations among basil taxa (Ocimum L.) based on molecular markers, nuclear DNA content, and chromosome number , 2009, Plant Systematics and Evolution.

[38]  M. Marchi,et al.  Genetic traceability of meat using microsatellite markers , 2008 .

[39]  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.

[40]  Jeremy R. deWaard,et al.  Biological identifications through DNA barcodes , 2003, Proceedings of the Royal Society of London. Series B: Biological Sciences.

[41]  M. Chase,et al.  Plastid rbc L sequence data indicate a close affinity between Diegodendron and Bixa , 1998 .

[42]  Gurcharan Singh Plant Systematics: An Integrated Approach , 1999 .

[43]  R. Hanner,et al.  DNA barcoding detects market substitution in North American seafood , 2008 .