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2019 Vol.31, Issue 3

Research Article

Development of Species-specific Primer Sets for Detecting Cirsium setidens and Silybum marianum using Real-time PCR Assay and Their Application in Commercial Food Products
J. An1
,
G. S. Kim1
,
C. S. Jang2*
1Master Students, Plant Genomics Laboratory, Department of Applied Plant Sciences, Kangwon National University
2Professor, Plant Genomics Laboratory, Department of Applied Plant Sciences, Kangwon National University
* Corresponding Author
31 December 2019. pp. 81-93
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Abstract

Cirsium setidens and Silybum marianum are medicinal plants that have been recently recognized as highly valuable sources of herbal drugs in Korea as well as worldwide. However, molecular markers for detecting these species in commercial food products have not yet been reported. In this study, we developed six species-specific primer sets for C. setidens and S. marianum using quantitative real-time PCR (qPCR) analysis, based on the single nucleotide polymorphisms (SNPs) of four chloroplast genes (ndhA, ndhF, rbcL and ycf1). The developed primer pairs showed correlation coefficients for their target species only (R2 > 0.99), and their efficiencies and slopes were in the range of 93－103% and -3.23 to -3.49, respectively. A total of 12 commercial food products including seven for C. setidens and 5 for S. marianum were tested to evaluate the feasibility of the developed primer sets. We found that C. setidens was present in all the seven C. setidens commercial products, while presence of S. marianum was confirmed in four of five S. marianum products. Therefore, the developed species-specific primer sets could prove a useful tool for detecting the presence of C. setidens and S. marianum in commercial food products.

Keywords
Cirsium setidens
Real-time PCR
Silybum marianum
SNP
Species-specific DNA markers
References
1
Abenavoli, L., Izzo, A., Milic, N., Cicala, C., Santini, A., Capasso, R. (2018) Milk thistle (Silybum marianum): A concise overview on its chemistry, pharmacological, and nutraceutical uses in liver diseases. Phytother Res 32:2202-2213.
10.1002/ptr.617130080294
2
Allmann, M., Candrian, U., Hoffelein, C., Luthy, J. (1993) Polymerase chain reaction (PCR): a possible alternative to immunochemical methods assuring safety and quality of food. Detection of wheat contamination in non-wheat food products. Eur Food Res Technol 196:248-251.
10.1007/BF012027418465611
3
An, J., Moon, J. C., Jang, C. S. (2018) Markers for distinguishing Orostachys species by SYBR Green-based real-time PCR and verification of their application in commercial O. japonica food products. Appl Biol Chem 61:499-508.
10.1007/s13765-018-0383-3
4
Arlorio, M., Cereti, E., Coisson, J. D., Travaglia, F., Martelli, A. (2007) Detection of hazelnut (Corylus spp.) in processed foods using real-time PCR. Food Control 18:140-148.
10.1016/j.foodcont.2005.09.005
5
Bahmani, M., Shirzad, H., Rafieian, S., Rafieian-Kopaei, M. (2015) Silybum marianum: beyond hepatoprotection. J Evidence-Based Complementary Altern Med 20:292-301.
10.1177/215658721557111625686616
6
Bustin, S. A., Benes, V., Garson, J. A., Hellemans, J., Huggett, J., Kubista, M., Mueller, R., Nolan, T., Pfaffl, M. W., Shipley, G. L., Vandesompele, J., Wittwer, C. T. (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55:611-622.
10.1373/clinchem.2008.11279719246619
7
Chen, S., Yu, H., Luo, H., Wu, Q., Li, C., Steinmetz, A. (2016) Conservation and sustainable use of medicinal plants: problems, progress, and prospects. Chin Med 11:37.
10.1186/s13020-016-0108-727478496PMC4967523
8
Curci, P. L., De Paola, D., Danzi, D., Vendramin, G. G., Sonnante, G. (2015) Complete chloroplast genome of the multifunctional crop globe artichoke and comparison with other Asteraceae. PLoS One 10 e0120589.
10.1371/journal.pone.012058925774672PMC4361619
9
Dong, W., Xu, C., Li, C., Sun, J., Zuo, Y., Shi, S., Cheng, T., Guo, J., Zhou, S. (2015) ycf1, the most promising plastid DNA barcode of land plants. Sci Rep 5:8348.
10.1038/srep0834825672218PMC4325322
10
ENGL (European Network of GMO Laboratories). Last accessed March 31, (2016) Definition of minimum performance requirements for analytical methods of GMO testing. http://gmo-crl.jrc.ec.europa.eu/doc/MPR%20Report%20 Application%2020_10_2015.pdf.
11
Garino, C., De Paolis, A., Coisson, J. D., Bianchi, D. M., Decastelli, L., Arlorio, M. (2016) Sensitive and specific detection of pine nut (Pinus spp.) by real-time PCR in complex food products. Food Chem 194:980-985.
10.1016/j.foodchem.2015.08.11426471643
12
Hayashi, K., Hashimoto, N., Daigen, M., Ashikawa, I. (2004) Development of PCR-based SNP markers for rice blast resistance genes at the piz locus. Theor Appl Genet 108:1212-1220.
10.1007/s00122-003-1553-014740086
13
Hirotsu, N., Murakami, N., Kashiwagi, T., Ujiie, K., Ishimaru, K. (2010) Protocol: A simple gel-free method for SNP genotyping using allele-specific primers in rice and other plant species. Plant Methods 6:12.
10.1186/1746-4811-6-1220409329PMC2876155
14
Jamil, A., Shahid, M., Khan, M. M., Ashraf, M. (2007) Screening of some medicinal plants for isolation of antifungal proteins and peptides. Pak J Bot (Pakistan) 39:211-221.
15
Jose Bagur, M., Alonso Salinas, G., Jimenez-Monreal, A., Chaouqi, S., Llorens, S., Martinez-Tome, M., Alonso, G. L. (2017) Saffron: An old medicinal plant and a potential novel functional food. Molecules 23:30.
10.3390/molecules2301003029295497PMC5943931
16
Khan, M. A., Abbasi, B. H., Shinwari, Z. K., (2014) Thidiazuron enhanced regeneration and silymarin content in Silybum marianum L. Pak J Bot 46:185-190.
10.1016/j.indcrop.2012.12.035
17
Kim, J. H., Hong, J. Y., Moon, J., Kwon, K., Jang, C. S. (2018) Development of molecular markers for detecting almond, peanut, pine nut, and walnut in commercial food using quantitative real-time PCR. Appl Biol Chem 61:345-354.
10.1007/s13765-018-0365-5
18
Kim, J. H., Moon, J., Kang, T. S., Kwon, K., Jang, C. S., (2017) Development of cpDNA markers for discrimination between Cynanchum wilfordii and Cynanchum auriculatum and their application in commercial C. wilfordii food products. Appl Biol Chem 60:79-86.
10.1007/s13765-017-0252-5
19
Kim, T., Lee, J., Jeong, G. H., Kim, T. H. (2016) Inhibitory effects of advanced glycation end products formation and free radical scavenging activity of Cirsium setidens. Korean J Food Preserv 23:283-289
10.11002/kjfp.2016.23.2.283
20
Lee, J. H., Jung, H. K., Han, Y., Yoon, Y. M., Yun, C. W., Sun, H. Y., Cho, H. W., Lee, S. H. (2016) Antioxidant effects of Cirsium setidens extract on oxidative stress in human mesenchymal stem cells. Mol Med Rep 14:3777-3784.
10.3892/mmr.2016.570627599894PMC5042755
21
Lee, S. H., Jin, Y. S., Heo, S. I., Shim, T. H., Sa, J. H., Choi, D. S., Wang, M. H. (2006) Composition analysis and antioxidative activity from different organs of Cirsium setidens Nakai. Korean J Food Sci Technol 38:571-576.
22
Lee, S., Lee, S. J., Kim, Y. (2018) Development of specific SNP molecular marker from Thistle using DNA sequences of ITS region. J Plant Biotechnol 45:102-109.
10.5010/JPB.2018.45.2.102
23
Li, S., Li, S., Gan, R., Song, F., Kuang, L., Li, H. (2013) Antioxidant capacities and total phenolic contents of infusions from 223 medicinal plants. Ind Crops Prod 51:289-298.
10.1016/j.indcrop.2013.09.017
24
Lo, Y. T., Shaw, P. C. (2018) DNA-based techniques for authentication of processed food and food supplements. Food Chem 240:767-774.
10.1016/j.foodchem.2017.08.02228946341
25
Pafundo, S., Gulli, M., Marmiroli, N. (2009) SYBR® GreenERTM real-time PCR to detect almond in traces in processed food. Food Chem 116:811-815.
10.1016/j.foodchem.2009.03.040
26
Ponchel, F., Toomes, C., Bransfield, K., Leong, F. T., Douglas, S. H., Field, S. L., Bell, S. M., Combaret, V., Puisieux, A., Mighell, A. J. (2003) Real-time PCR based on SYBR-green I fluorescence: An alternative to the TaqMan assay for a relative quantification of gene rearrangements, gene amplifications and micro gene deletions. BMC Biotechnol 3:18.
10.1186/1472-6750-3-1814552656PMC270040
27
Provan, J., Powell, W., Hollingsworth, P. M. (2001) Chloroplast microsatellites: new tools for studies in plant ecology and evolution. Trends Ecol Evol 16:142-147.
10.1016/S0169-5347(00)02097-8
28
Rajeswara, R. B. R., Syamasundar, K. V., Rajput, D. K., Nagaraju, G., Adinarayana, G. (2012) Biodiversity, conservation and cultivation of medicinal plants. J Pharmacogn 3:59-62.
29
Ramakers, C., Ruijter, J. M., Deprez, R. H. L., Moorman, A. F. (2003) Assumption-free analysis of quantitative real-time polymerase chain reaction (PCR) data. Neurosci Lett 339:62-66.
10.1016/S0304-3940(02)01423-4
30
Villa, C., Costa, J., Oliveira, MBP., Mafra, I. (2017) Novel quantitative real-time PCR approach to determine safflower (Carthamus tinctorius) adulteration in saffron (Crocus sativus). Food Chem 229:680-687.
10.1016/j.foodchem.2017.02.13628372231
Information
  • Publisher :Agriculture and Life Sciences Research Institute, Kangwon National University
  • Publisher(Ko) :강원대학교 농업생명과학대학 농업생명과학연구원
  • Journal Title :Journal of Agricultural, Life and Environmental Sciences
  • Journal Title(Ko) :농업생명환경연구
  • Volume : 31
  • No :3
  • Pages :81-93
  • Received Date : 2019-11-15
  • Revised Date : 2019-11-30
  • Accepted Date : 2019-11-30
  • DOI :https://doi.org/10.22698/jales.20190010
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