Abstract
The dried plant material of medicinally important Anthemis deserti Boiss. (family: Asteraceae) especially when it remains in the powdered form often look similar to Anthemis melampodina Del.; and therefore, difficult to distinguish, finally lead to chances of adulteration. The adulteration in medicinal plants effects on the efficacy of the drugs. The molecular authentication of herbal plant materials such as based on the internal transcribed spacer 2 (ITS2) sequences of nuclear ribosomal DNA (nrDNA) is considered as more reliable method compared to other the biochemical or histological methods. The present study aims to molecular authentication ofA. deserti based on molecular phylogenetic analyses of ITS2 gene sequence of nrDNA region. The ITS2 region of nrDNA of A. deserti were sequenced, and the molecular phylogenetic analyses were performed together with the GenBank sequences. The Maximum Parsimony tree revealed the close relationships of A. deserti with A. melampodina; however, the Neighbor-Joining and Maximum Likelihood tree clearly revealed that A. deserti is distinct from A. melampodina, which is also supported by the differences in nucleotides at five diffident positions (i.e. 22, 28, 87, 175 and 198) in the DNA sequence alignment.
Keywords: Molecular authentication, Anthemis deserti, Asteraceae, ITS2, nrDNA, Medicinal plants
1. Introduction
The genus Anthemis (family: Asteraceae) includes approximately 210 species (Bremer and Humphries, 1993) distributed in Europe, south west Asia, and north and north east Africa, is medicinally important genus as evident from the several pharmacological studies (Quarenghi et al., 2000, Stamatis et al., 2003, Uzel et al., 2004, Akgul and Saglikoglu, 2005, Buruk et al., 2006, Magro et al., 2006, Karioti et al., 2007, Karioti et al., 2008, Karioti et al., 2009, Papaioannou et al., 2007, Réthy et al., 2007, Collu et al., 2008, Di Giorgio et al., 2008, Hajdú et al., 2010, Vucković et al., 2010, Seol et al., 2010, Kilic et al., 2011, Conforti et al., 2012, Formisano et al., 2012, Samadi et al., 2012). In Saudi Arabia, the genus Anthemis is represented by 17 species viz., Anthemis arvensis L., A. bornmuelleri Stoj. & Acht., A. cotula L., A. deserti Boiss., A. dicksoniae Ghafoor, A. edumea Eig., A. haussknechtii Boiss. & Reut., A. hyalina DC., A. leptophylla Eig., A. melampodina Del., A. pseudocotula Boiss., A. rascheyana Boiss., A. scrobicularis Yavin, A. sheilae Ghafoor & Al-Turki, A. tenuicarpa Eig., A. yemenensis Podl. and A. zoharyana Eig. (Ghafoor and Al Turki, 2000). Among these, the extracts of A. melanolepis have been reported to have activity against Helicobacter pylori (Stamatis et al., 2003). The flavonoids from A. cotula flowers possess antimicrobial activity (Quarenghi et al., 2000). The essential oils of water-distilled vegetative parts of A. pseudocotula possess medicinally important molecules such as 1,8-cineole, camphor, artemisiaketone, filifolene (Kilic et al., 2011). A. arvensis and A. cotula possess pharmacologically important linear sesquiterpene lactones (Vucković et al., 2010).
The difficulties in discrimination of the species of the genus Anthemis is due to the diverse variability in morphological characters (Lo Presti et al., 2010). In the field condition, due to overlapping of the morphological characters, the medicinally important Anthemis speices need exhaustive taxonomic expertiese for txonomic identification especially to discriminate among A. cotula, A. melampodina, and A. pseudocotula. The dried plant material of A. deserti especially when it remains in powdered form often look similar to A. melampodina; and therefore, difficult to distinguish; finally lead to chances of adulteration (pers. obs.). The adulteration in medicinal plants effects on the efficacy of the drugs (Jayasinghe et al., 2009). The DNA sequence based molecular authentication of herbal plant materials such as based on the internal transcribed spacer 2 (ITS2) sequences of nuclear ribosomal DNA (nrDNA) is considered as more reliable in comparison to the biochemical or histological methods (Zhang et al., 2007, Kiran et al., 2010, Chen et al., 2010, Poczai and Hyvönen, 2010, Yao et al., 2010, Zuo et al., 2011, Ali et al., 2014). Hence, the present study aims to authentication of A. deserti based on ITS2 gene sequence of nrDNA region.
2. Materials and methods
2.1. Plant material and sequencing of ITS2 region of nrDNA
The leaves of Anthemis deserti were collected from Riyadh region of Saudi Arabia, and fixed in silica gel of 60–120 mesh fine silica gel powder. The herbarium voucher specimens were prepared for the references. The identification was checked by matching the vegetative and reproductive morphological characters of voucher specimen with taxonomic description mentioned in the Flora (Ghafoor and Al Turki, 2000). The voucher specimens were submitted to KSUH (Herbarium, King Saud University, Riyadh, Saudi Arabia) for the record and reference.
The total genomic DNA was isolated, which was then subjected to thermal cycling, the polymerase chain reactions in order to amplification of the nrDNA ITS2 region. The product of polymerase chain reactions was used for DNA sequencing following the method previously described (Ali et al., 2010, Ali et al., 2013a, Ali et al., 2013b, Ali et al., 2014, Ali et al., 2015, Ali et al., 2016, Al-Hemaid et al., 2014, Al-Hemaid et al., 2015, Choudhary et al., 2011).
2.2. Molecular phylogenetic analyses of nrDNA ITS2 region
The nrDNA ITS2 sequences of 11 species of Anthemis (i.e. A. arvensis, A. bornmuelleri, A. cotula, A. edumea, A. haussknechtii, A. hyaline, A. leptophylla, A. melampodina, A. pseudocotula, A. rascheyana and A. zoharyana) were retrieved from GenBank (Table 1), and trimming at both start and end position were performed according to span in order to include only ITS2 region in the phylogenetic analyses. The ITS2 sequence of Tripleurospermum transcaucasicum (GenBank AJ864612) was used as outgroup in the phylogenetic analyses. The DNA sequences were aligned using MUSCLE (Edgar, 2004) and the phylogenetic analyses were performed using Neighbor-Joining (NJ) method (Zuckerkand and Pauling, 1965, Saitou and Nei, 1987, Rzhetsky and Nei, 1992, Dopazo, 1994, Kumar et al., 2016), Maximum Parsimony (MP) method (Felsenstein, 1985, Nei and Kumar, 2000, Kumar et al., 2016) with 100 bootstrap replicates (Felsenstein, 1985) and Maximum Likelihood (ML) method based on the JTT matrix-based model (Jones et al., 1992, Kumar et al., 2016) using the software MEGA X (Kumar et al., 2018).
Table 1.
The Genbank accesion number of the taxon included in the molecular phylogenetic analyses.
| Taxon | GenBank accession number | ||
|---|---|---|---|
| Ingroup | |||
| 1. | Anthemis arvensis L. | MG218603 | |
| 2. | Anthemis bornmuelleri Stoj. & Acht. | FM957784 | |
| 3. | Anthemis cotula L. | AJ312823 | |
| 4. | Anthemus edumea Eig. | FM957692 | |
| 5. | Anthemis haussknechtii Boiss. & Reut. | FM957658 | |
| 6. | Anthemis hyalina DC. | AJ312808 | |
| 7. | Anthemis leptophylla Eig. | FM957738 | |
| 8. | Anthemis melampodina Del. | AJ312809 | |
| 9. | Anthemis pseudocotula Boiss. | AJ312824 | |
| 10. | Anthemis rascheyana Boiss. | FM957702 | |
| 11. | Anthemis scrobicularis Yavin | FM957755 | |
| 12. | Anthemis zoharyana Eig. | FM957714 | |
| Outgroup | |||
| 14. | Tripleurospermum transcaucasicum | AJ864612 | |
3. Results and discussion
The aligned DNA data matrix of ITS2 region of A. arvensis, A. bornmuelleri, A. cotula, A. deserti, A. edumea, A. haussknechtii, A. hyaline, A. leptophylla, A. melampodina, A. pseudocotula, A. rascheyana and A. zoharyana and the outgroup Tripleurospermum transcaucasicum were 205 base pair long. One out of the six most parsimonious trees (length = 75) showed consistency index (CI) 0.685 and retention index (RI) 0.792. The ITS2 region of A. deserti was 202 base pairs (GC content 46%). The MP tree (Fig. 1) revealed that A. deserti nested in a clade with A. melampodina and A. zoharyana with strong bootstrap support (BS 81%). Similar relationship ofA. deserti with A. melampodina and A. zoharyana were also recovered in NJ analyses (Fig. 2) and ML (Fig. 3) analysis (BS: 93% in NJ, 87% in ML analysis). The comparison of nrDNA ITS2 sequence of A. deserti with A. melampodina (Fig. 4) revealed that there are differences in nucleotides at five different position in the DNA sequence alignment (i.e. at the alignment position 22, 28, 87 and 175 there was ‘T’ in A. deserti but ‘C’ in A. melampodina, while at the alignment position 198 there was ‘G’ in A. melampodina but ‘R’ in A. deserti. Morphologically, the key characteristics of A. deserti are receptacle hemispherical, achenes obpyramidate, disc corollas inflated towards the base in fruit, which differs from the A. melampodina in having the morphological characteristics disc corollas inflated and indurated in fruit, achenes tuberculate, paleae keeled, not stiffy acuminate.
Fig. 1.
The maximum parsimony tree [tree #one out of six most parsimonious trees (length = 45), CI: 0.703, RI: 0.822].
Fig. 2.
The Evolutionary relationships of taxa inferred using the Neighbor-Joining method.
Fig. 3.
The Maximum Likelihood tree with the highest log likelihood (-557.00).
Fig. 4.
A comaprision of nueclotide diffeeences in between A. deserti and A. melampodina.
A number of molecular markers such as Restriction Fragment Length Polymorphisms (RFLPs) have been employed to distinguish Aegle marmelos, Desmodium giganicum, Oroxylum indicum, Solanum xanthocarpum, Solanum indicum, Tribulus terresteris (Biswas and Biswas, 2013), Boerhavia diffusa (Biswas et al., 2013) and Angelica species (Feng et al., 2010); Amplified Fragment Length Polymorphisms (AFLPs) for American ginseng (Hon et al., 2003), Capsicum species (Shirasawa et al., 2013); Randomly Amplified Polymorphic DNA (RAPD) for Clitoria ternatea (Ali et al., 2013a, Ali et al., 2013b), Convolvulus pluricaulis (Ganie et al., 2015), Evolvulus alsinoides (Ganie and Sharma, 2014); Simple Sequence Repeats (SSRs) for Aloe vera (Tripathi et al., 2011), Echinacea spp (Russi et al., 2009), Plectranthus (Passinho-Soares et al., 2006), Embelica ribes (Gowda et al., 2010); Inter Simple Sequence Repeats (ISSR) for Rheum species (Wang, 2011), Swertia (Tamhankar et al., 2009), Cissampelos pareira (Vijayan et al. 2014); Sequence Characterized Amplified Regions (SCAR) for Bacopa monnieri (Yadav et al., 2012), Aconitum heterophyllum and Cyperus rotundus (Seethapathy et al., 2014), Lonicera japonica (Fu et al., 2013), Ophiopogon japonicas (Li and Park, 2012), Phyllanthus amarus (Theerakulpisut et al., 2008); Loop Mediated Isothermal Amplification (LAMP) for Nigella sativa (Ganie et al., 2013), Taraxacum formosanum (Lai et al., 2015), Zingiber officinale (Chaudhary et al., 2014), Curcuma longa (Sasaki and Nagumo, 2007), Catharanthus roseus (Chaudhary et al., 2012), Panex ginseng (Sasaki et al. 2008). The latest advancement in the DNA sequencing technology and bioinformatics tools for DNA sequence data analysis lead to the development of DNA barcoding techniques (Hebert et al., 2003, Hebert et al., 2004, Hebert and Gregory, 2005, Hebert and Barrett, 2005, Ali et al., 2014) which have revolutionaries the method of the plant taxonomic identification (Poczai and Hyvönen, 2010) using DNA barcoding method especially based on DNA barcode sequence such as nrDNA ITS1, nrDNA ITS2 (Chen et al., 2010, Yao et al., 2010), rbcL, matK, ycf5, rpoC1, psbA-trnH, rps16, trnL-F and ndhF (Ali et al., 2014). DNA barcoding have successfully employed in authentication of Crocus sativus (Jiang et al., 2014), Schisandra chinensis (Li et al., 2013), Astragalus (Gao et al., 2009). The Next Generation Sequencing (NGS) is comparatively new, and have been demonstrated in the authentication of Costus pictus (Annadurai et al., 2012), Aconitum (Yun et al., 2015), Dendrobium officinale (Guo et al., 2013), Huperzia serrata and Phlegmariurus carinatus (Luo et al., 2010), Valeriana officinalis (Pyle et al., 2012), Hippophae rhamnoides (Ghangal et al., 2013), Ocimum sanctum (Rastogi et al., 2015), Beta vulgaris (Dohm et al., 2014), Panax ginseng (Jayakodi et al., 2014), Elaeis guineensis (Singh et al., 2013), Curcuma longa (Annadurai et al., 2013), Catharanthus roseus (Van Moerkercke et al., 2013), Withania somnifera (Gupta et al., 2013), Azadirachta indica (Krishnan et al., 2012), Cannabis sativa (van Bakel et al., 2011) and Populus trichocarpa (Tuskan et al., 2006). Moreover, Chen et al., (2010) demonstrated the potential use of ITS2 in the DNA barcoding of medicinal plants. The present species specific molecular signature of A. deserti nevertheless will be useful in molecular authentication, molecular phylogeny and DNA barcoding of the genus.
Acknowledgments
Research supported by the King Saud University, Deanship of Scientific Research, College of Science, Research Center.
Footnotes
Peer review under responsibility of King Saud University.
References
- Akgul C., Saglikoglu G. Antibacterial activity of crude methanolic extract and its fractions of aerial parts of Anthemis tinctoria. Ind. J. Biochem. Biophys. 2005;42(6):395–397. [PubMed] [Google Scholar]
- Al-Hemaid F.M.A., Ali M.A., Lee J., Gyulai G., Pandey A.K. Application of internal transcribed spacer of nuclear ribosomal DNA for identification of Echinops mandavillei Kit Tan. Bang. J. Pl. Tax. 2014;21(1):33–42. [Google Scholar]
- Al-Hemaid F.M.A., Ali M.A., Lee J., Kim S.Y., Rahman M.O. Molecular evolutionary relationships of Euphorbia Scordifolia Jacq. within the genus inferred from analysis of internal transcribed spacer sequences. Bang. J. Pl. Tax. 2015;22(2):111–118. [Google Scholar]
- Ali M.A., Van D.L., Kim S.K. Molecular systematic study of Cardamine glechomifolia Levl. (Brassicaceae) using internal transcribed spacer sequence of nuclear ribosomal DNA (ITS) and chloroplast trnL and trnL-F sequences. Saudi J. Biolog. Sci. 2010;17(4):275–290. doi: 10.1016/j.sjbs.2010.06.001. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ali M.A., Lee J., Kim S.Y., Park S.H., Al-Hemaid F.M.A. Molecular phylogenetic analyses of internal transcribed spacer sequences of nuclear ribosomal DNA defined monophyly of the genus Phytolacca L. (Phytolaccaceae) Bang. J. Pl. Tax. 2015;22(1):1–8. [Google Scholar]
- Ali M.A., Gábor G., Norbert H., Balázs K., Al-Hemaid F.M.A., Pandey A.K., Lee J. The changing epitome of species identification – DNA barcoding. Saudi J. Biol. Sci. 2014;21(3):204–231. doi: 10.1016/j.sjbs.2014.03.003. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ali M.A., Lee J., Oliur Rahman M., Al-Anazi F.S.M., Al-Hemaid F.M.A., Hatamleh A.A., Lee C., Mylliemngap B.J., Bhattacharjee A. A phylogenetic implication of molecular genotyping of Euryops jaberiana abedin & Chaudhary (Asteraceae) Bang. J. Pl. Tax. 2016;23(1):45–51. [Google Scholar]
- Ali M.A., Al-Hemaid F.M.A., Choudhary R.K., Lee J., Kim S.Y., Rub M.A. Status of Reseda pentagyna Abdallah & A.G. Miller (Resedaceae) inferred from analysis of combined nuclear ribosomal and chloroplast sequence data. Bang. J. Pl. Tax. 2013;20(2):233–238. [Google Scholar]
- Ali Z., Ganie S.H., Narula A., Sharma M.P., Srivastava P.S. Intra-specific genetic diversity and chemical profiling of different accessions of Clitoria ternatea L. Ind. Crop. Prod. 2013;43:768–773. [Google Scholar]
- Annadurai R.S., Jayakumar V., Mugasimangalam R.C., Katta M.A., Anand S., Gopinathan S., Sarma S.P., Fernandes S.J., Mullapudi N., Murugesan S., Rao S.N. Next generation sequencing and de novo transcriptome analysis of Costus pictus D. Don, a nonmodel plant with potent anti-diabetic properties. BMC Genomics. 2012;13:663/. doi: 10.1186/1471-2164-13-663. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Annadurai R.S., Neethiraj R., Jayakumar V., Damodaran A.C., Rao S.N., Katta M.A., Gopinathan S., Sarma S.P., Senthilkumar V., Niranjan V., Gopinath A., Mugasimangalam R.C. De novo transcriptome assembly (NGS) of Curcuma longa L. rhizome reveals novel transcripts related to anticancer and antimalarial terpenoids. PLoS. ONE. 2013;8:e56217. doi: 10.1371/journal.pone.0056217. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Biswas K., Biswas R. Identification ofmedicinal plants using PCR-RFLP in Dasamula — an Ayurvedic drug. J. Pharm. BioSci. 2013;3:94–99. [Google Scholar]
- Biswas K., Kapoor A., Biswas R. Authentication of herbal medicinal plant — Boerhavia diffusa L. Using PCR-RFLP. Curr. Trends Biotechnol. Pharm. 2013;7:716–1124. [Google Scholar]
- Bremer K., Humphries C.J. Generic monograph of the asteraceae-anthemideae. Bull. Nat. Hist. Mus. Lond. (Bot.) 1993;23(2):71–177. [Google Scholar]
- Buruk K., Sokmen A., Aydin F., Erturk M. Antimicrobial activity of some endemic plants growing in the eastern black sea region, Turkey. Fitoterapia. 2006;77(5):388–391. doi: 10.1016/j.fitote.2006.03.002. [DOI] [PubMed] [Google Scholar]
- Chaudhary A., Khan M., Al-Shaqha W.M., Alharbi M., Al-Khamees O.A. Rapid and easy molecular authentication of medicinal plant Zingiber officinale Roscoe by loop mediated isothermal amplification (LAMP)-based marker. J. Med. Plants. Res. 2014;8:756–762. [Google Scholar]
- Chaudhary A.A., Hemant, Mohsin M., Ahmad A. Application of loop-mediated isothermal amplification (LAMP)-based technology for authentication of Catharanthus roseus (L.) G. Don. Protoplasma. 2012;249:417–422. doi: 10.1007/s00709-011-0293-2. [DOI] [PubMed] [Google Scholar]
- 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 the ITS2 region as a novel DNA barcode for identifying medicinal plant species. PLoS One. 2010;5(1):e8613. doi: 10.1371/journal.pone.0008613. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Choudhary R.K., Ali M.A., Lee J. Studies on genetic diversity among population of Persicaria barbata (L.) Hara from India based on internal transcribed spacer sequences of nuclear ribosomal DNA. Saudi J. Biolog. Sci. 2011;18(2):123–127. doi: 10.1016/j.sjbs.2010.12.010. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Collu F., Bonsignore L., Casu M., Floris C., Gertsch J., Cottiglia F. New cytotoxic saturated and unsaturated cyclohexanones from Anthemis maritima. Bioorg. Med. Chem. Lett. 2008;18(5):1559–1562. doi: 10.1016/j.bmcl.2008.01.078. [DOI] [PubMed] [Google Scholar]
- Conforti F., Menichini F., Formisano C., Rigano D., Senatore F., Bruno M., Rosselli S., Celik S. Anthemis wiedemanniana essential oil prevents LPS-induced production of NO in RAW 264.7 macrophages and exerts antiproliferative and antibacterial activities in vitro. Nat. Prod. Res. 2012;26(17):1594–1601. doi: 10.1080/14786419.2011.585988. [DOI] [PubMed] [Google Scholar]
- Dohm J.C., Minoche A.E., Holtgräwe D., Capella-Gutiérrez S., Zakrzewski F., Tafer H., Rupp O., Sörensen T.R., Stracke R., Reinhardt R., Goesmann A., Kraft T., Schulz B., Stadler P.F., Schmidt T., Gabaldón T., Lehrach H., Weisshaar B., Himmelbauer H. The genome of the recently domesticated crop plant sugar beet (Beta vulgaris) Nature. 2014;505:546–549. doi: 10.1038/nature12817. [DOI] [PubMed] [Google Scholar]
- Di Giorgio C., Delmas F., Tueni M., Cheble E., Khalil T., Balansard G. Alternative and complementary antileishmanial treatments: assessment of the antileishmanial activity of 27 Lebanese plants, including 11 endemic species. J. Altern. Complement. Med. 2008;14(2):157–162. doi: 10.1089/acm.2007.7041. [DOI] [PubMed] [Google Scholar]
- Dopazo J. Estimating errors and confidence intervals for branch lengths in phylogenetic trees by a bootstrap approach. J. Mol. Evol. 1994;38:300–304. doi: 10.1007/BF00176092. [DOI] [PubMed] [Google Scholar]
- Edgar R.C. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res. 2004;32(5):1792–1797. doi: 10.1093/nar/gkh340. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Felsenstein J. Confidence limits on phylogenies: an approach using the bootstrap. Evolution. 1985;39:783–791. doi: 10.1111/j.1558-5646.1985.tb00420.x. [DOI] [PubMed] [Google Scholar]
- Feng T., Liu S., Xing-jin H. Molecular authentication of the traditional Chinese medicinal plant Angelica sinensis based on internal transcribed spacer of nrDNA. Electron. J. Biotechnol. 2010;13(1) [Google Scholar]
- Formisano C., Rigano D., Senatore F., Raimondo F.M., Maggio A., Bruno M. Essential oil composition and antibacterial activity of Anthemis mixta and A. tomentosa (Asteraceae) Nat. Prod. Commun. 2012;7(10):1379–1382. [PubMed] [Google Scholar]
- Fu J., Yang L., Khan M.A., Mei Z. Genetic characterization and authentication of Lonicera japonica Thunb by using improved RAPD analysis. Mol. Biol. Rep. 2013;40:5993–5999. doi: 10.1007/s11033-013-2703-3. [DOI] [PubMed] [Google Scholar]
- Ganie S.H., Ali Z., Das S., Srivastava P.S., Sharma M.P. Molecular characterization and chemical profiling of different populations of Convolvulus pluricaulis (Convolvulaceae); an important herb of Ayurvedic medicine. 3. Biotechnology. 2015;5:295–302. doi: 10.1007/s13205-014-0227-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ganie S.H., Sharma M.P. Molecular and chemical profiling of different populations of Evolvulus alsinoides (L.) L. Int. J. Agricul. Crop. Sci. 2014;7:1322–1331. [Google Scholar]
- Ganie S.H., Yadav D., Ahmad A., Chadhry A., Asif A. Authentication of traditional crop Kalongi (Nigella sativa L.) by LAMP marker. Ind. J. Res. Pharm. Biotechnol. 2013;1:765–771. [Google Scholar]
- Gao T., Pang X.H., Chen S.L. Authentication of plants in Astragalus by DNA barcoding technique. Planta Med. 2009;75:PB21. [Google Scholar]
- Ghafoor, A., Al Turki, T.A., 2000. Anthemis L. in: Chaudhary, Fl. Kingd. of Saudi Arabia, vol. 2, no. 3, pp. 168–176.
- Ghangal R., Chaudhary S., Jain M., Purty R.S., Sharma P.C. Optimization of de novo short read assembly of seabuckthorn (Hippophae rhamnoides L.) transcriptome. PLoS One. 2013;8:e72516. doi: 10.1371/journal.pone.0072516. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gowda B., Chandrika K., Prasanna K.T., Kirana V.C. Authentication of Embelica ribes Burm F. and Embelica tsjeriam-cottam A. DC. Int. J. Sci. Nat. 2010;1:58–60. [Google Scholar]
- Guo X., Li Y., Li C. Analysis of the Dendrobium officinale transcriptome reveals putative alkaloid biosynthetic genes and genetic markers. Gene. 2013;527:131–138. doi: 10.1016/j.gene.2013.05.073. [DOI] [PubMed] [Google Scholar]
- Gupta P., Goel R., Pathak S., Srivastava A., Singh S.P., Sangwan R.S., Asif M.H., Trivedi P.K. De novo assembly, functional annotation and comparative analysis of Withania somnifera leaf and root transcriptomes to identify putative genes involved in the withanolides biosynthesis. PLoS One. 2013;8:e62714. doi: 10.1371/journal.pone.0062714. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hajdú Z., Zupkó I., Réthy B., Forgo P., Hohmann J. Bioactivity-guided isolation of cytotoxic sesquiterpenes and flavonoids from Anthemis ruthenica. Planta Med. 2010;76(1):94–96. doi: 10.1055/s-0029-1185942. [DOI] [PubMed] [Google Scholar]
- Hebert P.D.N., Barrett R.D.H. Reply to the comment by L. Prendini on ‘‘Identifying spiders through DNA barcodes’’. Can. J. Zool. 2005;83:505–506. [Google Scholar]
- Hebert P.D.N., Cywinska A., Ball S.L., deWaard J.R. Biological identifications through DNA barcodes. Proc. Biol. Sci. 2003;270(1512):313–321. doi: 10.1098/rspb.2002.2218. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hebert P.D.N., Gregory T.R. The promise of DNA barcoding for taxonomy. Syst. Biol. 2005;54(5):852–859. doi: 10.1080/10635150500354886. [DOI] [PubMed] [Google Scholar]
- Hebert P.D.N., Stoeckle M.Y., Zemlak T.S., Francis C.M. Identification of birds through DNA barcodes. PLoS Biol. 2004;2(10) doi: 10.1371/journal.pbio.0020312. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hon C.C., Chow Y.C., Zeng F.Y., Leung F.C. Genetic authentication of ginseng and other traditional Chinese medicine. Acta Pharmacol. Sin. 2003;24:841–846. [PubMed] [Google Scholar]
- Jayakodi M., Lee S.C., Park H.S., Jang W., Choi B.S., Nah G.J., Kim D.S., Natesan S., Sun C., Lee Y.S., Yang T.J. Transcriptome profiling and comparative analysis of Panax ginseng adventitious roots. J. Ginseng Res. 2014;38:278–288. doi: 10.1016/j.jgr.2014.05.008. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jayasinghe R., Hai N.L., Coram T.E., Kong S., Kaganovitch J., Xue C.C., Li C.G., Pang E.C. Effectiveness of an innovative prototype subtracted diversity array (SDA) for fingerprinting plant species of medicinal importance. Planta Med. 2009;75:1180–1185. doi: 10.1055/s-0029-1185484. [DOI] [PubMed] [Google Scholar]
- Jiang C., Cao L., Yuan Y., Chen M., Jin Y., Huang L. Barcoding melting curve analysis for rapid, sensitive, and discriminating authentication of saffron (Crocus sativus L.) from its adulterants. BioMed. Res. 2014:809037. doi: 10.1155/2014/809037. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jones D.T., Taylor W.R., Thornton J.M. The rapid generation of mutation data matrices from protein sequences. Comput. Appl. Biosci. 1992;8:275–282. doi: 10.1093/bioinformatics/8.3.275. [DOI] [PubMed] [Google Scholar]
- Karioti A., Skaltsa H., Kaiser M., Tasdemir D. Trypanocidal, leishmanicidal and cytotoxic effects of anthecotulide-type linear sesquiterpene lactones from Anthemis auriculata. Phytomedicine. 2009;16(8):783–787. doi: 10.1016/j.phymed.2008.12.008. [DOI] [PubMed] [Google Scholar]
- Karioti A., Skaltsa H., Linden A., Perozzo R., Brun R., Tasdemir D. Anthecularin: a novel sesquiterpene lactone from Anthemis auriculata with antiprotozoal activity. J. Org. Chem. 2007;72(21):8103–8106. doi: 10.1021/jo701751w. [DOI] [PubMed] [Google Scholar]
- Karioti A., Skaltsa H., Zhang X., Tonge P.J., Perozzo R., Kaiser M., Franzblau S.G., Tasdemir D. Inhibiting enoyl-ACP reductase (FabI) across pathogenic microorganisms by linear sesquiterpene lactones from Anthemis auriculata. Phytomedicine. 2008;15(12):1125–1129. doi: 10.1016/j.phymed.2008.02.018. [DOI] [PubMed] [Google Scholar]
- Kilic O., Kocak A., Bagci E. Composition of the volatile oils of two Anthemis L. taxa from Turkey. Z Naturforsch C. 2011;66(11–12):535–540. doi: 10.1515/znc-2011-11-1201. [DOI] [PubMed] [Google Scholar]
- Kiran U., Khan S., Mirza K.J., Ram M., Abdin M.Z. SCAR markers: a potential tool for authentication of herbal drugs. Fitoterapia. 2010;81:969–976. doi: 10.1016/j.fitote.2010.08.002. [DOI] [PubMed] [Google Scholar]
- Krishnan N.M., Pattnaik S., Jain P., Gaur P., Choudhary R., Srividya V., Sa D., Arun K.H., Krishna P.G.B., Nair J., Varghese L., Valivarthi N.K., Dhas K., Ramaswamy K., Panda B. A draft of the genome and four transcriptomes of a medicinal and pesticidal angiosperm Azadirachta indica. BMC Genomics. 2012;13:464. doi: 10.1186/1471-2164-13-464. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kumar S., Stecher G., Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol. Biol. Evol. 2016;33:1870–1874. doi: 10.1093/molbev/msw054. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kumar S., Stecher G., Li M., Knyaz C., Tamura K. MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol. Biol. Evol. 2018;35(6):1547–1549. doi: 10.1093/molbev/msy096. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lai G.H., Chao J., Lin M.K., Chang W.T., Peng W.H., Sun F.C., Lee M.S., Lee M.S. Rapid and sensitive identification of the herbal tea ingredient Taraxacum formosanum using loop mediated isothermal amplification. Int. J. Mol. Sci. 2015;16:1562–1575. doi: 10.3390/ijms16011562. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Li G., Park Y.J. SCAR markers for discriminating species of two genera of medicinal plants, Liriope and Ophiopogon. Genet. Mol. Res. 2012;11:2987–2996. doi: 10.4238/2012.May.18.14. [DOI] [PubMed] [Google Scholar]
- Li X., Wang B., Han R., Zheng Y., Yin H.Y., Xu L. Identification of medicinal plant Schisandra chinensis using a potential DNA barcode ITS2. Acta Soc. Bot. Pol. 2013;82:283–288. [Google Scholar]
- Lo Presti R.M., Oppolzer S., Oberprieler C.H. A molecular phylogeny and a revised classification of the Mediterranean genus Anthemis s.l. (Compositae, Anthemideae) based on three molecular markers and micromorphological characters. Taxon. 2010;59:1441–1456. [Google Scholar]
- Luo H., Li Y., Sun C. Comparison of 454-ESTs from Huperzia serrata and Phlegmariurus carinatus reveals putative genes involved in lycopodium alkaloid biosynthesis and developmental regulation. BMC Plant Biol. 2010;10:209. doi: 10.1186/1471-2229-10-209. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Magro A., Carolino M., Bastos M., Mexia A. Efficacy of plant extracts against stored-products fungi. Rev Iberoam Micol. 2006;23(3):176–178. doi: 10.1016/s1130-1406(06)70039-0. [DOI] [PubMed] [Google Scholar]
- Nei M., Kumar S. Oxford University Press; New York: 2000. Molecular Evolution and Phylogenetics. [Google Scholar]
- Papaioannou P., Lazari D., Karioti A., Souleles C., Heilmann J., Hadjipavlou-Litina D., Skaltsa H. Phenolic compounds with antioxidant activity from Anthemis tinctoria L. (Asteraceae) Z Naturforsch C. 2007;62(5–6):326–330. doi: 10.1515/znc-2007-5-603. [DOI] [PubMed] [Google Scholar]
- Passinho-Soares H., Felix D., Kaplan M.A., Margis-Pinheiro M. Authentication of medicinal plant botanical identity by amplified fragmented length polymorphism dominant DNA marker: inferences from the Plectranthus genus. Planta Med. 2006;72:929–931. doi: 10.1055/s-2006-946673. [DOI] [PubMed] [Google Scholar]
- Poczai P., Hyvönen J. Nuclear ribosomal spacer regions in plant phylogenetics: problems and prospects. Mol. Biol. Rep. 2010;37(4):1897–1912. doi: 10.1007/s11033-009-9630-3. [DOI] [PubMed] [Google Scholar]
- Pyle B.W., Tran H.T., Pickel B., Haslam T.M., Gao Z., Macnevin G., Vederas J.C., Kim S.U., Ro D.K. Enzymatic synthesis of valerena-4,7(11)-diene by a unique sesquiterpene synthase from the valerian plant (Valeriana officinalis) FEBS J. 2012;279:3136–3146. doi: 10.1111/j.1742-4658.2012.08692.x. [DOI] [PubMed] [Google Scholar]
- Quarenghi M.V., Tereschuk M.L., Baigori M.D., Abdala L.R. Antimicrobial activity of flowers from Anthemis cotula. Fitoterapia. 2000;71(6):710–712. doi: 10.1016/s0367-326x(00)00229-x. [DOI] [PubMed] [Google Scholar]
- Rastogi S., Kalra A., Gupta V., Khan F., Lal R.K., Tripathi A.K., Parameswaran S., Gopalakrishnan C., Ramaswamy G., Shasany A.K. Unravelling the genome of Holy basil: an “incomparable” “elixir of life” of traditional Indian medicine. BMC Genomics. 2015;16:413. doi: 10.1186/s12864-015-1640-z. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Réthy B., Csupor-Löffler B., Zupkó I., Hajdú Z., Máthé I., Hohmann J., Rédei T., Falkay G. Antiproliferative activity of Hungarian Asteraceae species against human cancer cell lines. Part I. Phytother. Res. 2007;21(12):1200–1208. doi: 10.1002/ptr.2240. [DOI] [PubMed] [Google Scholar]
- Russi L., Moretti C., Raggi L., Albertini E., Falistocco E. Identifying commercially relevant Echinacea species by AFLP molecular markers. Genome. 2009;52:912–918. doi: 10.1139/g09-066. [DOI] [PubMed] [Google Scholar]
- Rzhetsky A., Nei M. A simple method for estimating and testing minimum evolution trees. Mol. Biol. Evol. 1992;9:945–967. [Google Scholar]
- Saitou N., Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 1987;4:406–425. doi: 10.1093/oxfordjournals.molbev.a040454. [DOI] [PubMed] [Google Scholar]
- Samadi N., Manayi A., Vazirian M., Samadi M., Zeinalzadeh Z., Saghari Z., Abadian N., Mozaffarian V.O., Khanavi M. Chemical composition and antimicrobial activity of the essential oil of Anthemis altissima L. var. altissima. Nat. Prod. Res. 2012;26(20):1931–1934. doi: 10.1080/14786419.2011.617750. [DOI] [PubMed] [Google Scholar]
- Sasaki Y., Komatsu K., Nagumo S. Rapid detection of Panax ginseng by loop mediated isothermal amplification and its application to authentication of ginseng. Biol. Pharm. Bull. 2008;31:1806–1808. doi: 10.1248/bpb.31.1806. [DOI] [PubMed] [Google Scholar]
- Sasaki Y., Nagumo S. Rapid identification of Curcuma longa and C. aromatica by LAMP. Biol. Pharm. Bull. 2007;30:2229–2230. doi: 10.1248/bpb.30.2229. [DOI] [PubMed] [Google Scholar]
- Seethapathy G.S., Balasubramani S.P., Venkatasubramanian P. NrDNA ITS sequence based SCAR marker to authenticate Aconitum heterophyllum and Cyperus rotundus in ayurvedic raw drug source and prepared herbal products. Food Chem. 2014;145:1015–1020. doi: 10.1016/j.foodchem.2013.09.027. [DOI] [PubMed] [Google Scholar]
- Seol G.H., Shim H.S., Kim P.J., Moon H.K., Lee K.H., Shim I., Suh S.H., Min S.S. Antidepressant-like effect of Salvia sclarea is explained by modulation of dopamine activities in rats. J. Ethnopharmacol. 2010;130(1):187–190. doi: 10.1016/j.jep.2010.04.035. [DOI] [PubMed] [Google Scholar]
- Shirasawa K., Ishii K., Kim C., Ban T., Suzuki M., Ito T., Muranaka T., Kobayashi M., Nagata N., Isobe S., Tabata S. Development of Capsicum EST-SSR markers for species identification and in silico mapping onto the tomato genome sequence. Mol. Breed. 2013;31:101–110. doi: 10.1007/s11032-012-9774-z. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Singh R., Ong-Abdullah M., Low E.T., Manaf M.A., Rosli R., Nookiah R., Ooi L.C., Ooi S.E., Chan K.L., Halim M.A., Azizi N., Nagappan J., Bacher B., Lakey N., Smith S.W., He D., Hogan M., Budiman M.A., Lee E.K., DeSalle R., Kudrna D., Goicoechea J.L., Wing R.A., Wilson R.K., Fulton R.S., Ordway J.M., Martienssen R.A., Sambanthamurthi R. Oil palm genome sequence reveals divergence of inter fertile species in old and new worlds. Nature. 2013;500:335–339. doi: 10.1038/nature12309. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stamatis G., Kyriazopoulos P., Golegou S., Basayiannis A., Skaltsas S., Skaltsa H. In vitro anti-Helicobacter pylori activity of Greek herbal medicines. J. Ethnopharmacol. 2003;88(2–3):175–179. doi: 10.1016/s0378-8741(03)00217-4. [DOI] [PubMed] [Google Scholar]
- Tamhankar S., Ghate V., Raut A., Rajput B. Molecular profiling of “Chirayat” complex using inter simple sequence repeat (ISSR) markers. Planta Med. 2009;75:1266–1270. doi: 10.1055/s-0029-1185543. [DOI] [PubMed] [Google Scholar]
- Theerakulpisut P., Kanawapee N., Maensiri D., Bunnag S., Chantaranothai P. Development of species-specific SCAR markers for identification of three medicinal species of Phyllanthus. J. Syst. Evol. 2008;46:614–621. [Google Scholar]
- Tripathi N., Saini N., Tiwari S. Assessment of genetic diversity among Aloe vera accessions using amplified fragment length polymorphism. Int. J. Med. Arom. Plants. 2011;1:115–121. [Google Scholar]
- Tuskan G.A., Difazio S., Jansson S., Bohlmann J., Grigoriev I., Hellsten U., Putnam N., Ralph S., Rombauts S., Salamov A., Schein J., Sterck L., Aerts A., Bhalerao R.R., Bhalerao R.P., Blaudez D., Boerjan W., Brun A., Brunner A., Busov V., Campbell M., Carlson J., Chalot M., Chapman J., Chen G.L., Cooper D., Coutinho P.M., Couturier J., Covert S., Cronk Q., Cunningham R., Davis J., Degroeve S., Déjardin A., Depamphilis C., Detter J., Dirks B., Dubchak I., Duplessis S., Ehlting J., Ellis B., Gendler K., Goodstein D., Gribskov M., Grimwood J., Groover A., Gunter L., Hamberger B., Heinze B., Helariutta Y., Henrissat B., Holligan D., Holt R., Huang W., Islam-Faridi N., Jones S., Jones-Rhoades M., Jorgensen R., Joshi C., Kangasjärvi J., Karlsson J., Kelleher C., Kirkpatrick R., Kirst M., Kohler A., Kalluri U., Larimer F., Leebens-Mack J., Leplé J.C., Locascio P., Lou Y., Lucas S., Martin F., Montanini B., Napoli C., Nelson D.R., Nelson C., Nieminen K., Nilsson O., Pereda V., Peter G., Philippe R., Pilate G., Poliakov A., Razumovskaya J., Richardson P., Rinaldi C., Ritland K., Rouzé P., Ryaboy D., Schmutz J., Schrader J., Segerman B., Shin H., Siddiqui A., Sterky F., Terry A., Tsai C.J., Uberbacher E., Unneberg P., Vahala J., Wall K., Wessler S., Yan G.G., Yin T., Douglas C., Marra M., Sandberg G., Van de Peer Y., Rokhsar D. The genome of black cottonwood, Populus trichocarpa (Torr. & Gray) Science. 2006;313:1596–1604. doi: 10.1126/science.1128691. [DOI] [PubMed] [Google Scholar]
- Uzel A., Guvensen A., Cetin E. Chemical composition and antimicrobial activity of the essential oils of Anthemis xylopoda O. Schwarz from Turkey. J. Ethnopharmacol. 2004;95(2–3):151–154. doi: 10.1016/j.jep.2004.06.034. [DOI] [PubMed] [Google Scholar]
- Van Bakel H., Stout J.M., Cote A.G., Tallon C.M., Sharpe A.G., Hughes T.R., Page J.E. The draft genome and transcriptome of Cannabis sativa. Genome Biol. 2011;12(10):R102. doi: 10.1186/gb-2011-12-10-r102. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Van Moerkercke A., Fabris M., Pollier J., Baart G.J., Rombauts S., Hasnain G., Rischer H., Memelink J., Oksman-Caldentey K.M., Goossens A. CathaCyc, a metabolic pathway database built from Catharanthus roseus RNA-Seq data. Plant Cell Physiol. 2013;54:673–685. doi: 10.1093/pcp/pct039. [DOI] [PubMed] [Google Scholar]
- Vijayan D., Cheethaparambil A., Pillai G.S., Balachandran I. Molecular authentication of Cissampelos pareira L. var. hirsuta (Buch.-Ham. ex DC.) Forman, the genuine source plant of ayurvedic raw drug ‘Patha’, and its other source plants by ISSR markers. 3. Biotech. 2014;4:559–562. doi: 10.1007/s13205-013-0183-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vucković I., Vujisić L., Stesević D., Radulović S., Lazić M., Milosavljević S. Cytotoxic guaianolide from Anthemis segetalis (Asteraceae) Phytother. Res. 2010;24(2):225–227. doi: 10.1002/ptr.2916. [DOI] [PubMed] [Google Scholar]
- Wang X.M. Inter-simple sequence repeats (ISSR) molecular finger printing markers for authenticating the genuine species of rhubarb. J. Med. Plant Res. 2011;5:758–764. [Google Scholar]
- Yadav A., Ahmad J., Chaudhary A.A., Altaf A. Development of sequence characterized amplified region (SCAR) marker for the authentication of Bacopa monnieri (L.) Wettst. Eur. J. Med. Plants. 2012;2:186–198. [Google Scholar]
- 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(10):e13102. doi: 10.1371/journal.pone.0013102. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yun Y.E., Yu J.N., Nam G.H., Ryu S.A., Kim S., Oh K., Lim C.E. Next-generation sequencing identification and characterization of microsatellite markers in Aconitum austrokoreense Koidz., an endemic and endangered medicinal plant of Korea. Genet. Mol. Res. 2015;14:4812–4817. doi: 10.4238/2015.May.11.13. [DOI] [PubMed] [Google Scholar]
- Zhang Y.B., Shaw P.C., Sze C.W., Wang Z.T., Tong Y. Molecular authentication of Chinese herbal materials. J. Food Drug Anal. 2007;15:1–9. [Google Scholar]
- Zuckerkand E., Pauling L. Academic Press; New York: 1965. Evolutionary divergence and convergence in proteins; pp. 97–166. [Google Scholar]
- Zuo Y., Chen Z., Kondo K., Funamoto T., Wen J., Zhou S. DNA barcoding of Panax species. Planta Med. 2011;77:182–187. doi: 10.1055/s-0030-1250166. [DOI] [PubMed] [Google Scholar]




