Journal of Natural Medicines

, Volume 72, Issue 2, pp 514–522 | Cite as

Morphological and genetic differences between Coptis japonica var. anemonifolia H. Ohba and Coptis japonica var. major Satake in Hokuriku area

  • Masashi Kitamura
  • Hirokazu Ando
  • Yohei SasakiEmail author
Original Paper


Coptis japonica is widely distributed in Japan, and its dried rhizome is a source of the domestic herbal medicine Coptidis Rhizoma (黄連 Oren). There are three varieties of C. japonica, two of which, namely, C. japonica var. anemonifolia and C. japonica var. major, are important as sources of traditional medicines. Coptis japonica var. anemonifolia and C. japonica var. major are distinguishable on the basis of their ternate or biternate compound leaves, respectively. In the Hokuriku area, where both C. japonica var. anemonifolia and C. japonica var. major grow naturally, some individual plants cannot be identified unambiguously on the basis of leaf morphology because changes in leaf morphology may occur due to intra-variety variation or crossbreeding between the two varieties. In addition, genetic differences between the two varieties have remained unclear. In this study, we employed new genetic and morphological classification approaches to discriminate between the two varieties. Based on the single nucleotide polymorphisms of the tetrahydroberberine oxidase gene, we found four conserved SNPs between the two varieties and were able to classify C. japonica into two varieties and crossbreeds. Furthermore, we introduced a new leaf type index based on the overall degree of leaflet dissection calculated by surface area of a leaflet and length of leaflet margin and petiolule. Using our new index we were able to discriminate between the two varieties and their crossbreeds more accurately than is possible with the conventional discrimination method. Our genetic and morphological classification methods may be used as novel benchmarks to discriminate between the two varieties and their crossbreeds.


Coptis japonica Tetrahydroberberine oxidase Coptidis Rhizoma Single-nucleotide polymorphism 



We are grateful to Ms. Kazumi Fukuhara, Mr. Naomi Kuwahata, and Mr. Yoshiaki Kato for kindly providing samples of C. japonica. We also thank Dr. Kozo Fukuda for providing useful information.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

11418_2018_1179_MOESM1_ESM.pdf (57 kb)
Supplementary material 1 (PDF 57 kb)


  1. 1.
    The Ministry of Health, Labour and Welfare (2016) The Japanese pharmacopoeia. 17th edn (English version). The Ministry of Health, Labour and Welfare, TokyoGoogle Scholar
  2. 2.
    Mikage M, Kawamoto M (1998) Herbological studies of Coptidis Rhizoma (1). On the reason why “Kaga-Oren” was estimated as of high quality in the Edo era. Nat Med 52(5):414–420Google Scholar
  3. 3.
    Satake Y (1949) A note on the Coptis of Japan. J Jpn Bot 24:69–74Google Scholar
  4. 4.
    Iwatsuki K, Boufford DE, Ohba H (2006) Flora of Japan. Kodansha, Tokyo,Google Scholar
  5. 5.
    Konishi T, Kiyosawa S, Shimada Y, Terakado S, Hosoido S (1998) Vegetation on some colonies of Coptis japonica in Makino-cho, Takashimagun, Pref. Shiga. Nat Med 52(4):375–383Google Scholar
  6. 6.
    Xiang KL, Wu SD, Yu SX, Liu Y, Jabbour F, Erst AS, Zhao L, Wang W, Chen ZD (2016) The first comprehensive phylogeny of Coptis (Ranunculaceae) and its implications for character evolution and classification. PLoS One 11:e0153127CrossRefGoogle Scholar
  7. 7.
    Corrado G, Piffanelli P, Caramante M, Coppola M, Rao R (2013) SNP genotyping reveals genetic diversity between cultivated landraces and contemporary varieties of tomato. BMC Genomics 14:835CrossRefGoogle Scholar
  8. 8.
    Mitsui Y, Setoguchi H (2012) Demographic histories of adaptively diverged riparian and non-riparian species of Ainsliaea (Asteraceae) inferred from coalescent analyses using multiple nuclear loci. BMC Evol Biol 12:254CrossRefGoogle Scholar
  9. 9.
    Okada N, Shinmyo A, Okada H, Yamada Y (1998) Purification and characterization of (S)-tetrahydroberberine oxidase from cultured Coptis japonica cells. Phytochemistry 27:979–982CrossRefGoogle Scholar
  10. 10.
    Matsushima Y, Minami H, Hori K, Sato F (2012) Pathway engineering of benzylisoquinoline alkaloid biosynthesis in transgenic California poppy cells with ectopic expression of tetrahydroberberine oxidase from Coptis japonica. Plant Biotechnol 29:473–481CrossRefGoogle Scholar
  11. 11.
    Kojoma M, Seki H, Yoshida S, Muranaka T (2006) DNA polymorphisms in the tetrahydrocannabinolic acid (THCA) synthase gene in “drug-type” and “fiber-type” Cannabis sativa L. Forensic Sci Int 159:132–140CrossRefGoogle Scholar
  12. 12.
    Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Molec Biol Evol 30(12):2725–2729CrossRefGoogle Scholar
  13. 13.
    Salgado-Salazar C, Rossman AY, Chaverri P (2013) Not as ubiquitous as we thought: taxonomic crypsis, hidden diversity and cryptic speciation in the cosmopolitan fungus Thelonectria discophora (Nectriaceae, Hypocreales, Ascomycota). PLoS One 8:e76737CrossRefGoogle Scholar
  14. 14.
    Sing T, Sander O, Beerenwinkel N, Lengauer T (2005) ROCR: visualizing classifier performance in R. Bioinformatics 21:3940–3941CrossRefGoogle Scholar
  15. 15.
    Robin X, Turck N, Hainard A, Tiberti N, Lisacek F, Sanchez JC, Müller M (2011) pROC: an open-source package for R and S + to analyze and compare ROC curves. BMC Bioinform 12:77CrossRefGoogle Scholar
  16. 16.
    Wang XM, Hou XQ, Zhang YQ, Li Y (2014) Morphological variation in leaf dissection of Rheum palmatum complex (Polygonaceae). PLoS One 9:e110760CrossRefGoogle Scholar
  17. 17.
    Jian H, Yang B, Zhang A, Zhang L, Xu X, Li J, Liu L (2017) Screening of candidate leaf morphology genes by integration of QTL mapping and RNA sequencing technologies in Oilseed Rape (Brassica napus L.). PLoS One 12:e0169641CrossRefGoogle Scholar
  18. 18.
    Viscosi V, Cardini A (2011) Leaf morphology, taxonomy and geometric morphometrics: a simplified protocol for beginners. PLoS One 6:e25630CrossRefGoogle Scholar
  19. 19.
    Li X, Li Y, Zhang Z, Li X (2015) Influences of environmental factors on leaf morphology of Chinese jujubes. PLoS One 10:e0127825CrossRefGoogle Scholar
  20. 20.
    Xu F, Guo W, Xu W, Wei Y, Wang R (2009) Leaf morphology correlates with water and light availability: what consequences for simple and compound leaves? Prog Nat Sci 19:1789–1798CrossRefGoogle Scholar

Copyright information

© The Japanese Society of Pharmacognosy and Springer Japan KK, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Laboratory of Molecular Pharmacognosy, Division of Pharmaceutical Sciences, Graduate School of Medical SciencesKanazawa UniversityKanazawaJapan

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