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Molecular Discrimination of Cynanchum wilfordii and Cynanchum auriculatum by InDel Markers of Chloroplast DNA

1
Jeonnam Institute of Natural Resources Research, Jangheung-gun, Jeollanam-do 59338, Korea
2
Department of Pharmacy, College of Pharmacy, Mokpo National University, Muan-gun, Jeollanam-do 58554, Korea
3
Department of Pharmaceutical Engineering, International University of Korea, Jinju-si, Gyeongsangnam-do 52833, Korea
*
Author to whom correspondence should be addressed.
Molecules 2018, 23(6), 1337; https://doi.org/10.3390/molecules23061337
Received: 15 May 2018 / Revised: 25 May 2018 / Accepted: 30 May 2018 / Published: 1 June 2018
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Abstract

The tuber of Cynanchum wilfordii (Baekshuoh Radix in Korean) is an important medicinal herb in Korea and China; however, it is difficult to differentiate C. wilfordii from a related medicinal herb, C. auriculatum (Baishouwu Radix in Chinese). We sought to develop a molecular method that could be used to distinguish between the tubers of C. wilfordii and C. auriculatum. We aligned the chloroplast genome sequences (available in the NCBI database) of the two species and identified three species-specific insertion and deletion (InDel) sites in the trnQ-psbK, rps2-rpoC2, and psaJ-rpl33 intergenic spacer (IGS) regions. To confirm the presence of these three InDels and validate their use as markers, we designed three primer pairs to amplify the trnQ-psbK, rps2-rpoC2, and psaJ-rpl33 IGS regions. Polymerase chain reaction (PCR) amplification of the trnQ-psbK IGS region yielded a 249 bp fragment for C. wilfordii, and 419 bp fragment for C. auriculatum, whereas the rps2-rpoC2 IGS primers produced a 629 bp fragment from C. wilfordii and a 282 bp fragment from C. auriculatum. In the psaJ-rpl33 IGS region, allele fragments of 342 and 360 bp in length were amplified from C. wilfordii, whereas 249 and 250 bp fragment were amplified from C. auriculatum. We propose these three InDel markers as a valuable, simple, and efficient tool for identifying these medicinal herbs and will thus reduce adulteration of these herbal materials in commercial markets. View Full-Text
Keywords: InDel markers; Cynanchum wilfordii Radix; Cynanchum auriculatum Radix; chloroplast genome sequence; Korean pharmacopoeia InDel markers; Cynanchum wilfordii Radix; Cynanchum auriculatum Radix; chloroplast genome sequence; Korean pharmacopoeia
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Kim, Y.; Choi, H.; Shin, J.; Jo, A.; Lee, K.-E.; Cho, S.-S.; Hwang, Y.-P.; Choi, C. Molecular Discrimination of Cynanchum wilfordii and Cynanchum auriculatum by InDel Markers of Chloroplast DNA. Molecules 2018, 23, 1337.

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