Sains Malaysiana 45(3)(2016): 323–327

Multiplex PCR Assays for Species Discrimination of Cymbopogon citratus (DC.) Stapf and C. nardus (L.) Rendle, Two Common ‘Serai’ (Lemon Grass) Species in Peninsular Malaysia

(Asai ‘Multiplex PCR’ bagi Membezakan Cymbopogon citratus (DC.) Stapf dan C. nardus (L.) Rendle, Dua Spesies Serai yang Biasa Diperoleh di Semenanjung Malaysia)

 

 

WEI LUN NG¹, SWEE KEONG YEAP², NUR SYAZANA MOHAMED ABU BAKAR¹,

WAN NURFATIN WAN MOHD JAAFAR¹ & SOON GUAN TAN¹*

 

¹Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences

Universiti Putra Malaysia, 43400 Serdang, Selangor Darul Ehsan, Malaysia

 

²Institut Biosains, Universiti Putra Malaysia, 43400 Serdang, Selangor Darul Ehsan, Malaysia

 

Received: 30 April 2015/Accepted: 28 September 2015

 

ABSTRACT

Aromatic grass species Cymbopogon citratus (‘serai biasa/serai makan’) and C. nardus (‘serai wangi’) can be commonly found throughout Peninsular Malaysia. C. citratus is used in traditional Malaysian cooking and brewed as tea, while C. nardus is used in traditional medicine for external application and in insect repellents. Due to similar morphologies, it can be difficult to tell apart the species at times. Based on DNA sequence alignments of three chloroplast DNA intergenic spacer regions, namely atpB-rbcL, trnH-psbA and trnL-trnF, we designed species-specific primers for multiplex PCR assays for rapid species discrimination between C. citratus and C. nardus. The method described here makes use of simple molecular techniques that are time- and cost-effective for large-scale application. Such an assay will be useful for the quality assurance of food and medicinal products.

 

Keywords: Food quality assurance; herb; molecular identification; serai makan; serai wangi

 

ABSTRAK

Spesies rumput aromatik Cymbopogon citratus (‘serai biasa/serai makan’) dan C. nardus (‘serai wangi’) biasa diperoleh di sekitar Semenanjung Malaysia. C. citratus digunakan dalam masakan tradisi Malaysia dan direbus sebagai teh, manakala C. nardus digunakan dalam perubatan tradisi untuk aplikasi luaran dan penghalau serangga. Oleh kerana morfologinya yang sama, kedua-dua spesies ini sukar untuk dibezakan. Berdasarkan penjajaran jujukan DNA daripada tiga ‘intergenic spacer’ DNA kloroplas iaitu atpB-rbcL, trnH-psbA dan trnL-trnF, ‘primer’ khusus telah dihasilkan untuk kaedah ‘multiplex PCR’ bagi membezakan C. citratus dan C. nardus dengan cepat. Kaedah yang digunakan melibatkan teknik molekul ringkas yang menjimatkan masa dan kos untuk penggunaan berskala besar. Kaedah ini berguna dalam proses penjaminan kualiti produk makanan dan ubat-ubatan.

 

Kata kunci: Herba; jaminan kualiti makanan; pengesahan secara molekul; serai makan; serai wangi

 

REFERENCES

 

Akhila, A. 2010. Essential Oil-Bearing Grasses: The genus Cymbopogon. Florida: CRC Press.

Chiang, T.Y., Schaal, B.A. & Peng, C.I. 1998. Universal primers for amplification and sequencing a noncoding spacer between the atpB and rbcL genes of chloroplast DNA. Bot. Bull. Acad. Sin. 39: 245-250.

Clarke, L.A., Rebelo, C.S., Goncalves, J., Boavida, M.G. & Jordan, P. 2001. PCR amplification introduces errors into mononucleotide and dinucleotide repeat sequences. J. Clin. Pathol.: Mol. Pathol. 54: 351-353.

Fajardo, V., Gonzalez, I., Rojas, M., Garcia, T. & Martin, R. 2010. A review of current PCR-based methodologies for the authentication of meats from game animal species. Trends in Food Sci. & Tech. 21(8): 408-421.

James, D., Schmidt, A., Wall, E., Green, M. & Masri, S. 2003. Reliable detection and identification of genetically modified maize, soybean, and canola by multiplex PCR analysis. J. Agric. Food Chem. 51(20): 5829-5834.

Kress, W.J., Wurdack, K.J., Zimmer, E.A., Weigt, L.A. & Janzen, D.H. 2005. Use of DNA barcodes to identify flowering plants. PNAS 102(23): 8369-8374.

Lin, W.F. & Hwang, D.F. 2007. A multiplex PCR assay for species identification of raw and cooked bonito. Food Control 19(9): 879-885.

Maia, M.F. & Moore, S.J. 2011. Plant-based insect repellents: a review of their efficacy, development and testing. Malaria Journal 10(Sup 1): S11.

Ng, W.L. & Szmidt, A.E. 2013. A simple and inexpensive molecular assay for species identification of Indo-West Pacific Rhizophora mangroves for conservation and management. Conservation Genet. Resour. 5: 1059-1061.

Settanni, L. & Corsetti, A. 2006. The use of multiplex PCR to detect and differentiate food- and beverage-associated microorganisms: A review. J. Microbiological Methods 69(1): 1-22.

Sultan, S.E. 2000. Phenotypic plasticity for plant development, function and life history. Trends in Plant Sci. 5(12): 537-542.

Taberlet, P., Gielly, L., Pautou, G. & Bouvet, J. 1991. Universal primers for amplification of three non-coding regions of chloroplast DNA. Plant Mol. Biol. 17: 1105-1109.

Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. & Kumar, S. 2011. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol. Biol. Evol. 28: 2731-2739.

Thompson, J.D., Higgins, D.G. & Gibson, T.J. 1994. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22: 4673-4680.

 

 

*Corresponding author; email: sgtan@upm.edu.my