, Volume 249, Issue 4, pp 1229–1237 | Cite as

Transcriptome analysis of 1- and 3-year-old Panax notoginseng roots and functional characterization of saponin biosynthetic genes DS and CYP716A47-like

  • Jian Li
  • Lan Ma
  • Shuting Zhang
  • Cailian Zuo
  • Na Song
  • Shusheng Zhu
  • Jinsong WuEmail author
Original Article


Main conclusion

Transcriptome analysis revealed high expression of saponin biosynthetic genes may account for highly accumulated saponins in 3-year-old Panax notoginseng roots and DS and CYP716A47 - like were functionally verified by transgenic tobacco.

Panax notoginseng is a well-known traditional medical herb that contains bioactive compounds known as saponins. Three major dammarene-type triterpene saponins including R1, Rb1, and Rg1 were found to be highly accumulated in the roots of 3-year-old plants when compared to those of 1-year-old plants. However, the underlying cellular mechanism is poorly understood. In this study, transcriptome analysis revealed that most genes involved in saponin biosynthesis in P. notoginseng roots augmented during their growth periods. The analysis of the KEGG pathway indicated that the primary metabolism, cell growth, and differentiation were less active in the roots of 3-year-old plant; however, secondary metabolisms were enhanced, thus providing molecular evidence for the harvesting of P. notoginseng roots in the 3rd year of growth. Furthermore, the functional role of DS and CYP716A47-like, two of the candidate genes involved in saponin biosynthesis isolated from P. notoginseng, were verified via overexpression in cultivated tobacco. Approximately, 0.325 µg g−1 of dammarenediol-II and 0.320 µg g−1 of protopanaxadiol were recorded in the dry leaves of transgenic tobacco overexpressed with DS and both DS and CYP716A47-like, respectively. This study provides insights into the molecular mechanisms for saponin accumulation in P. notoginseng roots during its growth period and paves a promising way to produce dammarenediol-II and protopanaxadiol via transgenic techniques.


1- and 3-year-old Panax notoginseng Dammarene-type triterpene saponins RNA-seq Transgenic tobacco 



High-performance liquid chromatography


Mevalonic acid


Acetyl-CoA acetyltransferase


3-Hydroxy-3-methylglutaryl coenzyme-A synthase


HMG-CoA reductase


Mevalonate kinase


Phosphomevalonate kinase


Mevalonate diphosphate decarboxylase


Isopentenyl diphosphate isomerase


Geranylgeranyl pyrophosphate synthase


Farnesyl diphosphate synthase


Squalene synthase


Squalene epoxidase


Dammarenediol-II synthase


Beta-amyrin synthase


Cycloartenol synthase


Coding sequence


Differentially expressed genes


Transcription factors


Wild type



We thank Biological Technology Open Platform of Kunming Institute of Botany, the Chinese Academy of Sciences for greenhouse services. This project is supported by the Major Science and Technique Programs in Yunnan Province (No. 2016ZF001), and 100-Oversea-Top-Talents Recruitment plan of Yunnan.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

425_2018_3083_MOESM1_ESM.pdf (112 kb)
Supplementary material 1 (PDF 112 kb)
425_2018_3083_MOESM2_ESM.xlsx (24 kb)
Supplementary material 2 (XLSX 23 kb)
425_2018_3083_MOESM3_ESM.tif (130 kb)
Supplementary material 3 Detection of introduced genes in transgenic tobacco overexpressing DS and CYP716A47-like. a Confirmation of the introduced genes DS by real-time PCR. b Expression of DS and CYP716A47-like in leaves of wild-type Nicotiana tobacum (WT) and DS/CYP transgenic lines (the DS and CYP716A47-like co-overexpressed lines) were detected by real-time PCR. Data are mean values with the standard deviation obtained from three independent plants. Not Detected (ND) means under detection limits (TIFF 129 kb)
425_2018_3083_MOESM4_ESM.tif (41 kb)
Supplementary material 4 Relative transcripts of genes GUS and DS in leaves of wild-type Nicotiana tobacum (WT), empty vector transgenic line (EV) and DS transgenic line 1 (DS 1#) were detected by real-time PCR. Data are mean values with the standard deviation obtained from three independent plants. The asterisks indicate significant differences between WT and other samples (Paired t test: **P < 0.01). Not Detected (ND) means under detection limits (TIFF 41 kb)


  1. Akihisa T, Tokuda H, Ukiya M, Suzuki T, Enjo F, Koike K, Nikaido T, Nishino AH (2004) 3-Epicabraleahydroxylactone and other triterpenoids from Camellia Oil and their inhibitory effects on Epstein–Barr virus activation. Chem Pharm Bull 52(1):153–156CrossRefGoogle Scholar
  2. Chun JH, Adhikari PB, Park SB, Han JY, Choi YE (2015) Production of the dammarene sapogenin (protopanaxadiol) in transgenic tobacco plants and cultured cells by heterologous expression of PgDDS and CYP716A47. Plant Cell Rep 34(9):1551–1560. CrossRefGoogle Scholar
  3. Han JY, Kim HJ, Kwon YS, Choi YE (2011) The Cyt P450 enzyme CYP716A47 catalyzes the formation of protopanaxadiol from dammarenediol-II during ginsenoside biosynthesis in Panax ginseng. Plant Cell Physiol 52(12):2062–2073. CrossRefGoogle Scholar
  4. Han JY, Hwang HS, Choi SW, Kim HJ, Choi YE (2012) Cytochrome P450 CYP716A53v2 catalyzes the formation of protopanaxatriol from protopanaxadiol during ginsenoside biosynthesis in Panax ginseng. Plant Cell Physiol 53(9):1535–1545. CrossRefGoogle Scholar
  5. Han JY, Wang HY, Choi YE (2014) Production of dammarenediol-II triterpene in a cell suspension culture of transgenic tobacco. Plant Cell Rep 33(2):225–233. CrossRefGoogle Scholar
  6. Haralampidis K, Trojanowska M, Osbourn AE (2002) Biosynthesis of triterpenoid saponins in plants. Adv Biochem Eng Biotechnol 75:31–49Google Scholar
  7. Jia XH, Wang CQ, Liu JH, Li XW, Wang X, Shang MY, Cai SQ, Zhu S, Komatsu K (2013) Comparative studies of saponins in 1-3-year-old main roots, fibrous roots, and rhizomes of Panax notoginseng, and identification of different parts and growth-year samples. J Nat Med 67(2):339–349. CrossRefGoogle Scholar
  8. Komakine N, Okasaka M, Takaishi Y, Kawazoe K, Murakami K, Yamada Y (2005) New dammarane-type saponin from roots of Panax notoginseng. J Nat Med 60(2):135–137. CrossRefGoogle Scholar
  9. Lee MH, Han JY, Kim HJ, Kim YS, Huh GH, Choi YE (2012) Dammarenediol-II production confers TMV tolerance in transgenic tobacco expressing Panax ginseng dammarenediol-II synthase. Plant Cell Physiol 53(1):173–182. CrossRefGoogle Scholar
  10. Li J, Wang J, Wu X, Liu D, Li J, Li J, Liu S, Gao W (2017) Jasmonic acid and methyl dihydrojasmonate enhance saponin biosynthesis as well as expression of functional genes in adventitious roots of Panax notoginseng F. H. Chen. Biotechnol Appl Biochem 64(2):225–238. CrossRefGoogle Scholar
  11. Liu MH, Yang BR, Cheung WF, Yang KY, Zhou HF, Kwok JS, Liu GC, Li XF, Zhong S, Lee SM, Tsui SK (2015) Transcriptome analysis of leaves, roots and flowers of Panax notoginseng identifies genes involved in ginsenoside and alkaloid biosynthesis. BMC Genom 16:265. CrossRefGoogle Scholar
  12. Ng TB (2006) Pharmacological activity of sanchi ginseng (Panax notoginseng). J Pharm Pharmacol 58(8):1007–1019. CrossRefGoogle Scholar
  13. Niu Y, Luo H, Sun C, Yang TJ, Dong L, Huang L, Chen S (2014) Expression profiling of the triterpene saponin biosynthesis genes FPS, SS, SE, and DS in the medicinal plant Panax notoginseng. Gene 533(1):295–303. CrossRefGoogle Scholar
  14. Sun H, Hu X, Ma J, Hettenhausen C, Wang L, Sun G, Wu J, Wu J (2014) Requirement of ABA signalling-mediated stomatal closure for resistance of wild tobacco to Alternaria alternata. Plant Pathol 63:1070–1107CrossRefGoogle Scholar
  15. Tansakul P, Shibuya M, Kushiro T, Ebizuka Y (2006) Dammarenediol-II synthase, the first dedicated enzyme for ginsenoside biosynthesis, in Panax ginseng. FEBS Lett 580(22):5143–5149. CrossRefGoogle Scholar
  16. Usami Y (2008) Antitumor agents. 261. 20(S)-protopanaxadiol and 20(S)-protopanaxatriol as antiangiogenic agents and total assignment of 1H NMR spectra. J Nat Prod 71(3):478–481CrossRefGoogle Scholar
  17. Wang C-Z, McEntee E, Wicks S, Wu J-A, Yuan C-S (2006) Phytochemical and analytical studies of Panax notoginseng (Burk.) F.H. Chen. J Nat Med 60:97–106. CrossRefGoogle Scholar
  18. Wang X-Y, Wanga D, Maa X-X, Yang Y-J, Za C-R (2008) Two new dammarane-type bisdesmosides from the fruit pedicels of Panax notoginseng. Helv Chim Acta 91:60–66CrossRefGoogle Scholar
  19. Wang JR, Yau LF, Gao WN, Liu Y, Yick PW, Liu L, Jiang ZH (2014) Quantitative comparison and metabolite profiling of saponins in different parts of the root of Panax notoginseng. J Agric Food Chem 62(36):9024–9034. CrossRefGoogle Scholar
  20. Wei G, Dong L, Yang J, Zhang L, Xu J, Yang F, Cheng R, Xu R, Chen S (2018) Integrated metabolomic and transcriptomic analyses revealed the distribution of saponins in Panax notoginseng. Acta Pharm Sin B 8(3):458–465. CrossRefGoogle Scholar
  21. Wu Q, Ma X, Zhang K, Feng X (2015) Identification of reference genes for tissue-specific gene expression in Panax notoginseng using quantitative real-time PCR. Biotechnol Lett 37:197–204. CrossRefGoogle Scholar
  22. Xia P, Guo H, Ru M, Yang D, Liang Z, Yan X, Liu Y (2017) Accumulation of saponins in Panax notoginseng during its growing seasons. Ind Crops Prod 104:287–292. CrossRefGoogle Scholar
  23. Yang M, Zhang X, Xu Y, Mei X, Jiang B, Liao J, Yin Z, Zheng J, Zhao Z, Fan L, He X, Zhu Y, Zhu S (2015) Autotoxic ginsenosides in the rhizosphere contribute to the replant failure of Panax notoginseng. PLoS One 10(2):e0118555. CrossRefGoogle Scholar
  24. Zhang D, Li W, Xia EH, Zhang QJ, Liu Y, Zhang Y, Tong Y, Zhao Y, Niu YC, Xu JH, Gao LZ (2017) The medicinal herb Panax notoginseng genome provides insights into ginsenoside biosynthesis and genome evolution. Mol Plant 10(6):903–907. CrossRefGoogle Scholar
  25. Zhou J, Huang W, Wu M, Yang C, Feng G, Wu Z (1975) Triterpenoids from Panax Linn. and their relationship with taxonomy and geographical distribution. Acta Phytotaxon Sin 13(2):41Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Yunnan Key Laboratory for Wild Plant ResourcesKunming Institute of Botany, Chinese Academy of SciencesKunmingChina
  2. 2.University of Chinese Academy of SciencesBeijingChina
  3. 3.Key Laboratory of Agro-Biodiversity and Pest Management of Education Ministry of China, State Key Laboratory for Conservation and Utilization of Bio-Resources in YunnanYunnan Agricultural UniversityKunmingChina

Personalised recommendations