Development of cpDNA markers for discrimination between Cynanchum wilfordii and Cynanchum auriculatum and their application in commercial C. wilfordii food products

• Published in: Applied biological chemistry Vol. 60; no. 1; pp. 79 - 86
• Main Authors: Kim, Ju Hee, Moon, Jun-Cheol, Kang, Tae Sun, Kwon, Kisung, Jang, Cheol Seong
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01-02-2017; Springer Nature B.V; 한국응용생명화학회
• Subjects: Applied Microbiology; Binary mixtures; Biological Techniques; Biomarkers; Bioorganic Chemistry; Chemistry; Chemistry and Materials Science; Chloroplasts; Correlation coefficients; Cultivation; Cynanchum; Deoxyribonucleic acid; DNA; Food; Food production; Food products; Nucleotides; Single-nucleotide polymorphism; Species; 농학; Quantitative real-time PCR; cpDNA marker; Commercial products
• ISSN: 2468-0834; 2468-0842
• DOI: 10.1007/s13765-017-0252-5

Cynanchum wilfordii
has been used as a health-enhancing food product in Korea. Owing to its morphological similarity to
C. wilfordii
and relative ease of cultivation,
C. auriculatum
has been illegally used as a substitute plant for
C. wilfordii
. In the present study, we developed markers to discriminate between
C. wilfordii
and
C. auriculatum
. Species-specific primer sets for
C. wilfordii
and
C. auriculatum
were designed based on the single nucleotide polymorphisms of the chloroplast
matK
genes for quantitative real-time PCR using SYBR green. The limit of detection of primer sets for each species was assessed by analyzing serially 1/10-diluted DNAs at concentrations of 0.001–100% (=10 ng) and binary mixtures of a flour matrix spiked with decreasing concentrations (10
3
–1 mg/g) of non- and heated
C. wilfordii
and
C. auriculatum
, respectively. We found that species-specific primer sets indicated good amplification efficiency and correlation coefficients (
R
2
) of the standard curves in the extracted DNA. The developed markers were successfully applied to 19 commercial
C. wilfordii
food products and could prove a useful tool for verifying the presence of
C. wilfordii
and
C. auriculatum
in commercial products.