The loci recommended as universal barcodes for plants on the basis of floristic studies may not work with congeneric species as exemplified by DNA barcoding of Dendrobium species

Hemant Kumar Singh1, Iffat Parveen2, Saurabh Raghuvanshi3, Shashi B. Babbar2
1Department of Botany, University of Delhi, Delhi-, India
2Department of Botany, University of Delhi, Delhi, India
3Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi, India

Tóm tắt

Abstract Background Based on the testing of several loci, predominantly against floristic backgrounds, individual or different combinations of loci have been suggested as possible universal DNA barcodes for plants. The present investigation was undertaken to check the applicability of the recommended locus/loci for congeneric species with Dendrobium species as an illustrative example. Results Six loci, matK, rbcL, rpoB, rpoC1, trnH-psbA spacer from the chloroplast genome and ITS, from the nuclear genome, were compared for their amplification, sequencing and species discrimination success rates among multiple accessions of 36 Dendrobium species. The trnH-psbA spacer could not be considered for analysis as good quality sequences were not obtained with its forward primer. Among the tested loci, ITS, recommended by some as a possible barcode for plants, provided 100% species identification. Another locus, matK, also recommended as a universal barcode for plants, resolved 80.56% species. ITS remained the best even when sequences of investigated loci of additional Dendrobium species available on the NCBI GenBank (93, 33, 20, 18 and 17 of ITS, matK, rbcL, rpoB and rpoC1, respectively) were also considered for calculating the percent species resolution capabilities. The species discrimination of various combinations of the loci was also compared based on the 36 investigated species and additional 16 for which sequences of all the five loci were available on GenBank. Two-locus combination of matK+rbcL recommended by the Plant Working Group of Consortium for Barcoding of Life (CBOL) could discriminate 86.11% of 36 species. The species discriminating ability of this barcode was reduced to 80.77% when additional sequences available on NCBI were included in the analysis. Among the recommended combinations, the barcode based on three loci - matK, rpoB and rpoC1- resolved maximum number of species. Conclusions Any recommended barcode based on the loci tested so far, is not likely to provide 100% species identification across the plant kingdom and thus is not likely to act as a universal barcode. It appears that barcodes, if based on single or limited locus(i), would be taxa specific as is exemplified by the success of ITS among Dendrobium species, though it may not be suitable for other plants because of the problems that are discussed.

Từ khóa


Tài liệu tham khảo

Hebert PDN, Cywinska A, Ball SL, de Waard JR: Biological identifications through DNA barcodes. Proc R Soc Biol Sci Ser B. 2003, 270: 313-321.

Hebert PDN, Ratnasingham S, de Waard JR: Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proc R Soc Biol Sci Ser B. 2003, 270: S96-S99.

Hebert PDN, Stoeckle MY, Zemlak TS, Francis CM: Identification of birds through DNA barcodes. PLoS Biol. 2004, 2: e312-

Hebert PDN, Penton EH, Burns JM, Janzen DH, Hallwachs W: Ten species in one: DNA barcoding reveals cryptic species in neotropical skipper butterfly Astraptes fulgerator. Proc Natl Acad Sci USA. 2004, 101: 14812-14817.

Lahaye R, van der Bank M, Bogarin D, Warner J, Pupulin F, Gigot G, Maurin O, Duthoit S, Barraclough TG, Savolainen V: DNA barcoding the floras of biodiversity hotspots. Proc Natl Acad Sci USA. 2008, 105: 2923-2928.

Parveen I, Singh HK, Raghuvanshi S, Pradhan UC, Babbar SB: DNA barcoding of endangered Indian Paphiopedilum species. Mol Ecol Resour. 2012, 12: 82-90.

Ragupathy S, Newmaster SG, Murugesan M, Balasubramaniam V: DNA barcoding discriminates a new cryptic grass species revealed in an ethnobotany study by the hill tribes of the Western Ghats in southern India. Mol Ecol Resour. 2009, 9: 164-171.

Newmaster SG, Ragupathy S: Testing plant barcoding in a sister species complex of pantropical Acacia (Mimosoideae, Fabaceae). Mol Ecol Resour. 2009, 9: 172-180.

Miwa H, Odrzykoski IJ, Matsui A, Hasegawa M, Akiyama H, Jia Y, Sabirov R, Takahashi H, Boufford DE, Murakami N: Adaptive evolution of rbcL in Conocephalum (Hepaticae, bryophytes). Gene. 2009, 441: 169-175.

Li FW, Tan BC, Buchbender V, Moran RC, Rouhan G, Wang CN, Quandt D: Identifying a mysterious aquatic fern gametophyte. Plant Syst Evol. 2009, 281: 77-86.

Chen S, Yao H, Han J, Liu C, Song J, Shi L, Zhu Y, Ma X, Gao T, Pang X, Luo K, Li Y, Li X, Jia X, Lin Y, Leon C: Validation of ITS2 region as a novel DNA barcode for identifying medicinal plant species. PLoS One. 2010, 5: 8613-

Valentini A, Miquel C, Taberlet P: DNA barcoding for honey biodiversity. Diversity. 2010, 2: 610-617.

Srirama R, Senthilkumar U, Sreejayan N, Ravikanth G, Gurumurthy BR, Shivanna MB, Sanjappa M, Ganeshaiah KN, Uma Shaanker R: Assessing species admixtures in raw drug trade of Phyllanthus, a hepato-protective plant using molecular tools. J Ethnopharmacol. 2010, 130: 208-215.

Ferri G, Alù M, Corradini B, Beduschi G: Forensic botany: species identification of botanical trace evidence using a multigene barcoding approach. Int J Legal Med. 2009, 123: 395-401.

Bleeker W, Klausmeyer S, Peintinger M, Dienst M: DNA sequences identify invasive alien Cardamine at Lake Constance. Biol Conserv. 2008, 141: 692-698.

Muellner AN, Schaefer H, Lahaye R: Evaluation of candidate DNA barcoding loci for economically important timber species of the mahogany family (Meliaceae). Mol Ecol Resour. 2011, 11: 450-460.

Yesson C, Bárcenas RT, Hernández HM, de la Luz Riz-Maqueda M, Prado A, Rodríguez VM, Hawkins JA: DNA barcodes for Mexican Cactaceae, plants under pressure from wild collecting. Mol Ecol Resour. 2011, 11: 775-783.

Soininen EM, Valentini A, Coissac E, Miquel C, Gielly L, Brochmann C, Brysting AK, Sønstebø JH, Ims RA, Yoccoz NG, Taberlet P: Analysing diet of small herbivores: the efficiency of DNA barcoding coupled with high-throughput pyrosequencing for deciphering the composition of complex plant mixtures. Front Zool. 2009, 6: 16-

Stech M, Kolvoort E, Loonen MJJE, Verieling K, Kruijer JD: Bryophyte DNA sequences from faeces of an arctic herbivore, barnacle goose (Branta leucopsis). Mol Ecol Resour. 2011, 11: 404-408.

Kress WJ, Wurdack KJ, Zimmer EA, Weigt LA, Janzen DH: Use of DNA barcodes to identify flowering plants. Proc Natl Acad Sci USA. 2005, 102: 8369-8374.

Newmaster SG, Fazekas AJ, Ragupathy S: DNA barcoding in land plants: evaluation of rbcL in a multigene tiered approach. Can J Bot. 2006, 84: 335-341.

Chase MW, Salamin N, Wilkinson M, Dunwell JM, Kesanakurthi RP, Haidar N, Savolainen V: Land plants and DNA barcodes: short-term and long-term goals. Philos Trans R Soc Lond B Biol Sci. 2005, 360: 1889-1895.

Kress WJ, Erickson DL: A two-locus global DNA barcode for land plants: the coding rbcL gene complements the non-coding trnH-psbA spacer region. PLoS One. 2007, 2: 508-

Sass C, Little DP, Stevenson DW, Specht CD: DNA Barcoding in the Cycadales: testing the potential of proposed barcoding markers for species identification of Cycads. PLoS One. 2007, 2: 1154-

Fazekas AJ, Burgess KS, Kesanakurti PR, Graham SW, Newmaster SG, Husband BC, Percy DM, Hajibabaei M, Barrett SCH: Multiple multilocus DNA barcodes from the plastid genome discriminate plant species equally well. PLoS One. 2008, 3: 2802-

Seberg O, Petersen G: How many loci does it take to DNA barcode a Crocus?. PLoS One. 2009, 4: 4598-

CBOL Plant Working Group: A DNA barcode for land plants. Proc Natl Acad Sci USA. 2009, 106: 12794-12797.

Yao H, Song JY, Ma XY, Liu C, Li Y, Xu HX, Han JP, Duan LS, Chen SL: Identification of Dendrobium species by a candidate DNA barcode sequence: the chloroplast psbA-trnH intergenic region. Planta Med. 2009, 75: 667-669.

He J, Wong KL, Shaw PC, Wang H, Li DZ: Identification of the medicinal plants in Aconitum L. by DNA barcoding technique. Planta Med. 2010, 76: 1622-1628.

Bruni I, de Mattia F, Galimberti A, Galasso G, Banfi E, Casiraghi M, Labra M: Identification of poisonous plants by DNA barcoding approach. Int J Legal Med. 2010, 124: 595-603.

Li F-W, Kuo L-Y, Rothfels CJ, Ebihara A, Chiou W-L, Windham MD, Pryer KM: rbcL and matK earn two thumbs up as the core DNA barcode for ferns. PLoS One. 2011, 6 (10): 26597-

Hollingsworth PM, Graham SW, Little DP: Choosing and using a plant DNA barcode. PLoS One. 2011, 6: 19254-

Chase MW, Cowan RS, Hollingsworth PM, van den Berg C, Madrinan S, Petersen G, Seberg O, Jorgsensen T, Cameron KM, Carine M, Pedersen N, Hedderson TAJ, Conrad F, Salazar GA, Richardson JE, Hollingsworth ML, Barraclough TG, Kelly L, Wilkinson M: A proposal for a standardized protocol to barcode all land plants. Taxon. 2007, 56: 295-299.

Roy S, Tyagi A, Shukla V, Kumar A, Singh UM, Chaudhary BL, Datt B, Bag SK, Singh PK, Nair NK, Husain T, Tuli R: Universal plant DNA barcode loci may not work in complex groups: a case study with Indian Berberis species. PLoS One. 2010, 5: 13674-

Nock CJ, Waters DLE, Edwards MA, Bowen SG, Rice N, Cordeiro GM, Henry RJ: Chloroplast genome sequences from total DNA for plant identification. Plant Biotechnol J. 2011, 9: 328-333.

Bulpitt CJ, Li Y, Bulpitt PF, Wang J: The use of orchids in Chinese medicine. J R Soc Med. 2007, 100: 558-563.

Chang CC, Lin HC, Lin IP, Chow TY, Chen HH, Chen WH, Cheng CH, Lin CY, Liu SM, Chang CC, Chaw SM: The chloroplast genome of Phalaenopsis aphrodite (Orchidaceae): comparative analysis of evolutionary rate with that of grasses and its phylogenetic implications. Mol Biol Evol. 2006, 23: 279-291.

Tamura K, Dudley J, Nei M, Kumar S: MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol. 2007, 24: 1596-1599.

Raspé O: Inheritance of the chloroplast genome in Sorbus aucuparia L. (Rosaceae). J Hered. 2001, 92: 507-509.

Wolfe K, Li WH, Sharp PM: Rates of nucleotide substitution vary greatly among plant mitochondrial, chloroplast, and nuclear DNAs. Proc Natl Acad Sci USA. 1987, 84: 9054-9058.

Schuiteman A, Bonnet P, Svengsuksa B, Barthelemy D: An annotated checklist of the Orchidaceae of Laos. Nord J Bot. 2008, 26: 257-316.

Alvarez I, Wendel JF: Ribosomal ITS sequences and plant phylogenetic inference. Mol Phylogenet Evol. 2003, 29: 417-434.

Bailey CD, Carr TG, Harris SA, Hughes CE: Characterization of angiosperm nrDNA polymorphism, paralogy, and pseudogenes. Mol Phylogenet Evol. 2003, 29: 435-455.

Gonzalez MA, Baraloto C, Engel J, Mori SA, Pétronelli P, Riéra B, Roger A, Thébaud C, Chave J: Identification of amazonian trees with DNA barcodes. PLoS One. 2009, 4: 7483-

Gao T, Yao H, Song J, Zhu Y, Liu C, Chen S: Evaluating the feasibility of using candidate DNA barcodes in discriminating species of the large Asteraceae family. BMC Evol Biol. 2010, 10: 324-

Gao T, Yao H, Song J, Liu C, Zhu Y, Ma X, Pang X, Xu H, Chen S: Identification of medicinal plants in the family Fabaceae using a potential DNA barcode ITS2. J Ethnopharmacol. 2010, 130: 116-121.

Pang X, Song J, Zhu Y, Xie C, Chen S: Using DNA barcoding to identify species within Euphorbiaceae. Planta Med. 2010, 76: 1784-1786.

Yao H, Song J, Liu C, Luo K, Han J, Li Y, Pang X, Xu H, Zhu Y, Xiao P, Chen S: Use of ITS2 region as the universal DNA barcode for plants and animals. PLoS One. 2010, 5: 13102-

Zhu YJ, Chen SL, Yao H, Tan R, Song JY, Luo K, Lu J: DNA barcoding the medicinal plants of the genus Paris. Yao Xue Xue Bao. 2010, 45: 376-382.

Whitlock BA, Hale AM, Groff PA: Intraspecific inversions pose a challenge for the trnH-psbA plant DNA barcode. PLoS One. 2010, 5: e11533-

Kress WJ, Erickson DL, Jones FA, Swensond NG, Perez R, Sanjur O, Bermingham E: Plant DNA barcodes and a community phylogeny of a tropical forest dynamics plot in Panama. Proc Natl Acad Sci USA. 2009, 106: 18621-18626.

Kress WJ, Erickson DL, Swenson NG, Thompson J, Uriarte M, Zimmerman JK: Advances in the use of DNA barcodes to build a community phylogeny for tropical trees in a Puerto Rican forest dynamics plot. PLoS One. 2010, 5: 15409-

Piredda R, Simeone MC, Attimonelli M, Bellarosa R, Schirone B: Prospects of barcoding the Italian wild dendroflora: oaks reveal severe limitations to tracking species identity. Mol Ecol Resour. 2011, 11: 72-83.

Burgess KS, Fazekas AJ, Kesanakurti PR, Graham SW, Husband BC, Newmaster SG, Percy DM, Hajibabaei M, Barrett SCH: Discriminating plant species in a local temperate flora using the rbcL + matK DNA barcode. Method Ecol Evol. 2011, 2: 333-340.

Ebihara A, Nitta JH, Ito M: Molecular species identification with rich floristic sampling: DNA barcoding the pteridophyte flora of Japan. PLoS One. 2010, 5 (12): 15136-

Asahina H, Shinozaki J, Masuda K, Morimitsu Y, Satake M: Identification of medicinal Dendrobium species by phylogenetic analyses using matK and rbcL sequences. J Nat Med. 2010, 64: 133-138.

Parks M, Cronn R, Liston A: Increasing phylogenetic resolution at low taxonomic levels using massively parallel sequencing of chloroplast genomes. BMC Biol. 2009, 7: 84-

Tate JA, Simpson BB: Paraphyly of Tarasa (Malvaceae) and diverse origins of the polyploid species. Syst Bot. 2003, 28: 723-737.

Tsai CC, Peng CI, Huang SC, Huang PL, Chou CH: Determination of the genetic relationship of Dendrobium species (Orchidaceae) in Taiwan based on the sequence of the internal transcribed spacer of ribosomal DNA. Sci Hortic. 2004, 101: 315-325.

Doyle JJ, Doyle JL: A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull. 1987, 19: 11-15.

Barnwell P, Blanchard AN, Bryant JA, Smirnoff N, Weir AF: Isolation of DNA from the highly mucilaginous succulent plant Sedum telephium. Plant Mol Biol Reptr. 1998, 16: 133-138.