Abstract
In this study, loop-mediated isothermal amplification (LAMP)-based molecular marker was developed for authentication of Catharanthus roseus, a medicinal plant. Samples of this plant were collected from different geographical locations in India. Random amplified polymorphic deoxyribonucleic acid (DNA) analysis of collected samples was carried out with 25 random primers. A 610-bp DNA fragment, common to all accessions, was eluted, cloned, and sequenced. Four LAMP primers were designed on the basis of sequence of 610 bp DNA fragment. LAMP reaction, containing 10× Bst DNA polymerase reaction buffer, Bst DNA polymerase, four in-house designed primers, dNTPs, MgSO4, and betaine, was incubated at 65°C for 1 h. The resulting amplicon was visualized by adding SYBR Green I to the reaction tube. The data showed confirmatory results. Since the assay method is simple, sensitive, and cost-effective, it is a feasible method for identifying and authentication of C. roseus.
Similar content being viewed by others
References
Angiolillo A, Mencuccini M, Baldoni L (1999) Olive genetic diversity assessed using amplified fragment length polymorphisms. Theor Appl Genet 98:411–421
Belaj A, la Trujillo I, Rosa R, Rallo L (2001) Polymorphism and discrimination capacity of randomly amplified polymorphic markers in an Olive germplasm bank. J Am Soc Hortic Sci 126:64–71
Carriero F, Fontanazza G, Cellini F, Giorio G (2002) Identification of simple sequence repeats (SSRs) in olive (Olea europea). Theor Appl Genet 104:301–307
Cipriani G, Marrazzo MT, Marconi R, Cimato A, Testolin R (2002) Microsatellite markers isolated in olive (Olea europea) are suitable for individual fingerprinting and reveal polymorphism within ancient cultivars. Theor Appl Genet 104:223–228
Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15
Ernst E (2005) The efficacy of herbal medicine—an overview. Fundam Clin Pharmacol 19:405–409
Ernst E (2006) Herbal medicines—they are popular, but are they also safe? Eur J Clin Pharmacol 62:1–2
Fabbri A, Hrmaza JI, Polito VS (1995) Random amplified polymorphic DNA analysis of Olive (Olea europea L.) cultivars. J Am Soc Hortic Sci 120:538–542
Hafner GJ, Yang IC, Wolter LC, Stafford MR, Giffard PM (2001) Isothermal amplification and multimerization of DNA by Bst DNA polymerase. Biotechniques 30:852–867
Hill J, Beriwal S, Chandra I, Paul VK, Kapil A, Singh T, Wadowsky MR, Singh V, Goyal A, Jahnukainen T, Johnson JR, Tarr IP, Vats A (2008) Loop-mediated isothermal amplification assay for rapid detection of common strains of Escherichia coli. J Clin Microbiol 46:2800–2804
Jones WP, Chin YW, Kinghorn AD (2006) The role of pharmacognosy in modern medicine and pharmacy. Curr Drug Targets 7:247–264
Koehn FE, Carter GT (2005) The evolving role of natural products in drug discovery. Nat Rev Drug Discov 4:206–220
McClelland M, Mathieu-Daude F, Welsh J (1995) RNA fingerprinting and differential display using arbitrarily primed PCR. Trends Gene 11:242–246
Mishra P, Kumar S (2000) Emergence of periwinkle C. roseus as a model system for molecular biology of alkaloids: phytochemistry, pharmacology, plant biology and in vivo and in vitro cultivation. J Med Aromat Plant Sci 22:306–337
Mizukami H, Ohbayashi K, Kitamura Y, Ikenaga T (1993) Restriction fragment length polymorphisms (RFLPs) of medicinal plants and crude drugs: I. RFLP probes allow clear identification of Duboisia interspecific hybrid genotypes in both fresh and dried tissues. Biol Pharm Bull 16:388–390
Mori YK, Nagamine N, Tomita T, Notori T (2001) Detection of loop mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem Biophys Res Commun 289:150–154
Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T (2000) Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28:E63
Rallo P, Dorado G, Martin A (2000) Development of simple sequence repeats (SSRs) in olive tree (Olea europea). Theor Appl Genet 101:984–989
Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory, New York
Sefc KM, Lopes MS, Mendonca D, Rodrigues Dos Santos M, Laimer Da Camara Machado M, Da Camara MA (2000) Identification of microsatellite loci in olive (Olea europea) and their characterization in Italian and Iberian olive trees. Mol Ecol 9:1171–1193
Zhao Z, Hu Y, Liang Z, Yuen JP, Jiang Z, Leung KS (2006) Authentication is fundamental for standardization of Chinese medicines. Planta Med 72:865–874
Conflicts of Interest
None
Author information
Authors and Affiliations
Corresponding author
Additional information
Handling Editor: Peter Nick
Rights and permissions
About this article
Cite this article
Chaudhary, A.A., Hemant, Mohsin, M. et al. Application of loop-mediated isothermal amplification (LAMP)-based technology for authentication of Catharanthus roseus (L.) G. Don. Protoplasma 249, 417–422 (2012). https://doi.org/10.1007/s00709-011-0293-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00709-011-0293-2