Skip to main content
Log in

Mining genes associated with furanocoumarin biosynthesis in an endangered medicinal plant, Glehnia littoralis

  • Research Article
  • Published:
Journal of Genetics Aims and scope Submit manuscript

Abstract

The endangered medicinal plant Glehnia littoralis is one of the important natural source of furanocoumarin, which has been used as mucolytic, antitussive, antitumour and antibacterial. However, the genetic information of furanocoumarin biosynthesis in G. littoralis is scarce at present. The objective of this study was to mine the putative candidate genes involved in the biosynthesis pathway of furanocoumarin and provide references for gene identification, and functional genomics of G. littoralis. We carried out the transcriptome analysis of leaves and roots in G. littoralis, which provided a dataset for gene mining. Psoralen, imperatorin and isoimperatorin were detected in G. littoralis by high performance liquid chromatography analysis. Candidate key genes were mined based on the annotations and local BLAST with homologous sequences using BioEdit software. The relative expression of genes was analysed using quantitative real-time polymerase chain reaction. Further, the CYP450 genes were mined using phylogenetic analyses using MEGA 6.0 software. A total of 156,949 unigenes were generated, of which 9021 were differentially-expressed between leaves and roots. A total of 82 unigenes encoding eight enzymes in furanocoumarin biosynthetic pathway were first obtained. Seven genes that encoded key enzymes in the downstream furanocoumarin biosynthetic pathway and expressed more in roots than leaves were screened. Twenty-six candidate CYP450 unigenes expressed abundantly in roots and were chiefly concentrated in CYP71, CYP85 and CYP72 clans. Finally, we filtered 102 differentially expressed transcription factors (TFs) unigenes. The transcriptome of G. littoralis was characterized which would help to elucidate the furanocoumarin biosynthetic pathway in G. littoralis and provide an invaluable resource for further study of furanocoumarin.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  • Andersen T. B., Martinezswatson K. A., Rasmussen S. A., Boughton B. A., Jørgensen K., Andersenranberg J. et al. 2017 Localization and in-vivo characterization of Thapsia garganica CYP76AE2 indicates a role in thapsigargin biosynthesis. Plant Physiol. 174, 56–72.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Bak S., Beisson F., Bishop G., Hamberger B., Hofer R., Paquette S. et al. 2011 Cytochromes P450. Arabidopsis Book 9, e0144.

    PubMed  PubMed Central  Google Scholar 

  • Bourgaud F., Hehn A., Larbat R., Doerper S., Gontier E., Kellner S. et al. 2006 Biosynthesis of coumarins in plants: a major pathway still to be unravelled for cytochrome P450 enzymes. Phytochem. Rev. 5, 293–308.

    CAS  Google Scholar 

  • Chen J., Dong X., Li Q., Zhou X., Gao S., Chen R. et al. 2013 Biosynthesis of the active compounds of Isatis indigotica based on transcriptome sequencing and metabolites profiling. BMC Genomics 14, 857.

    PubMed  PubMed Central  Google Scholar 

  • Chen S., Wang Y., Yang Y., Xiang T., Liu J., Zhou H. and Wu X. 2017 Psoralen inhibited apoptosis of osteoporotic osteoblasts by modulating IRE1-ASK1-JNK pathway. Bio. Med. Res. Int. 2017, 3524307.

    Google Scholar 

  • Cherukupalli N., Divate M., Mittapelli S. R., Khareedu V. R. and Vudem D. R. 2016 De novo assembly of leaf transcriptome in the medicinal plant Andrographis paniculata. Front. Plant Sci. 7, 1203.

    PubMed  PubMed Central  Google Scholar 

  • Chezem W. R., Memon A., Li F. S., Weng J. K. and Clay N. K. 2017 SG2-Type R2R3-MYB transcription factor MYB15 controls defense-induced lignification and basal immunity in arabidopsis. Plant Cell 29, 1907–1926.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Chinese Pharmacopoeia Commission 2015 Pharmacopoeia of the People’s Republic of China. Chinese Medical Science Press, Beijing.

    Google Scholar 

  • Costa V., Angelini C., De Feis I. and Ciccodicola A. 2010 Uncovering the complexity of transcriptomes with RNA-Seq. J. Biomed. Biotechnol. 2010, 853916.

    PubMed  PubMed Central  Google Scholar 

  • Field B. and Osbourn A. E. 2008 Metabolic diversification-independent assembly of operon-like gene clusters in different plants. Science 320, 543–547.

    CAS  PubMed  Google Scholar 

  • Field B., Fiston Lavier A. S., Kemen A., Geisler K., Quesneville H. and Osbourn A. E. 2011 Formation of plant metabolic gene clusters within dynamic chromosomal regions. Proc. Natl. Acad. Sci. USA 108, 16116.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Fu L. K. and Jin J. M. 1991 China plant red data book. Science Press, Beijing.

    Google Scholar 

  • Gorelick J. and Bernstein N. 2014 Chapter five-elicitation: an under utilized tool in the development of medicinal plants as a source of therapeutic secondary metabolites. Adv. Agron. 124, 201–230.

    CAS  Google Scholar 

  • Guo J., Zhou Y. J., Hillwig M. L., Shen Y., Yang L., Wang Y. et al. 2013 CYP76AH1 catalyzes turnover of miltiradiene in tanshinones biosynthesis and enables heterologous production of ferruginol in yeasts. Proc. Natl. Acad. Sci. USA 110, 12108–12113.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Guo J., Ma X., Cai Y., Ma Y., Zhan Z., Zhou Y. J. et al. 2016 Cytochrome P450 promiscuity leads to a bifurcating biosynthetic pathway for tanshinones. New Phytol. 210, 525–534.

    CAS  PubMed  Google Scholar 

  • Hehmann M., Lukacin R., Ekiert H. and Matern U. 2004 Furanocoumarin biosynthesis in Ammi majus L. Cloning of bergaptol O-methyltransferase. Eur. J. Biochem. 271, 932–940.

    CAS  PubMed  Google Scholar 

  • Hung W. L., Suh J. H. and Wang Y. 2017 Chemistry and health effects of furanocoumarins in grapefruit. J. Food Drug Anal. 25, 71–83.

    CAS  PubMed  Google Scholar 

  • Ishikawa A., Kitamura Y., Ozeki Y., Itoh Y., Yamada A. and Watanabe M. 2005 Post-stress metabolism involves umbelliferone production in anthocyanin-producing and nonproducing cells of Glehnia littoralis suspension cultures. J. Plant Physiol. 162, 703–710.

    CAS  PubMed  Google Scholar 

  • Ishikawa A., Kuma T., Sasaki H., Sasaki N., Ozeki Y., Kobayashi N. et al. 2009 Constitutive expression of bergaptol O-methyltransferase in Glehnia littoralis cell cultures. Plant Cell Rep. 28, 257–265.

    CAS  PubMed  Google Scholar 

  • Itkin M., Rogachev I., Alkan N., Rosenberg T., Malitsky S., Masini L. et al. 2011 Glycoalkaloid metabolism1 is required for steroidal alkaloid glycosylation and prevention of phytotoxicity in tomato. Plant Cell 23, 4507–4525.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Itkin M., Heinig U., Tzfadia O., Bhide A. J., Shinde B., Cardenas P. D. et al. 2013 Biosynthesis of antinutritional alkaloids in solanaceous crops is mediated by clustered genes. Science 341, 175–179.

    CAS  PubMed  Google Scholar 

  • Ji A., Jia J., Xu Z., Li Y., Bi W., Ren F. et al. 2017 Transcriptome-guided mining of genes involved in crocin biosynthesis. Front. Plant Sci. 8, 518.

    PubMed  PubMed Central  Google Scholar 

  • Kang U., Han A. R., So Y., Jin C. H., Ryu S. M., Lee D. et al. 2019 Furanocoumarins from the roots of angelica dahurica with inhibitory activity against intracellular reactive oxygen species accumulation. J. Nat. Prod. 82, 9, 2601–2607.

    CAS  PubMed  Google Scholar 

  • Karamat F., Olry A., Munakata R., Koeduka T., Sugiyama A., Paris C. et al. 2014 A coumarin-specific prenyltransferase catalyzes the crucial biosynthetic reaction for furanocoumarin formation in parsley. Plant J. 77, 627–638.

    CAS  PubMed  Google Scholar 

  • Kruse T., Ho K., Yoo H. D., Johnson T., Hippely M., Park J. H. et al. 2008 In planta biocatalysis screen of P450s identifies 8-methoxypsoralen as a substrate for the CYP82C subfamily, yielding original chemical structures. Chem. Biol. 15, 149–156.

    CAS  PubMed  Google Scholar 

  • Larbat R., Kellner S., Specker S., Hehn A., Gontier E., Hans J. et al. 2007 Molecular cloning and functional characterization of psoralen synthase, the first committed monooxygenase of furanocoumarin biosynthesis. J. Biol. Chem. 282, 542–554.

    CAS  PubMed  Google Scholar 

  • Larbat R., Hehn A., Hans J., Schneider S., Jugde H., Schneider B. et al. 2009 Isolation and functional characterization of CYP71AJ4 encoding for the first P450 monooxygenase of angular furanocoumarin biosynthesis. J. Biol. Chem. 284, 4776–4785.

    CAS  PubMed  Google Scholar 

  • Legay S., Lacombe E., Goicoechea M., Brière C., Séguin A., Mackay J. et al. 2007 Molecular characterization of EgMYB1, a putative transcriptional repressor of the lignin biosynthetic pathway. Plant Sci. 173, 542–549.

    CAS  Google Scholar 

  • Lin C. L., Hsiao G., Wang C. C. and Lee Y. L. 2016 Imperatorin exerts antiallergic effects in Th2-mediated allergic asthma via induction of IL-10-producing regulatory T cells by modulating the function of dendritic cells. Pharmacol. Res. 110, 111–121.

    CAS  PubMed  Google Scholar 

  • Liu J., Osbourn A. and Ma P. 2015 MYB transcription factors as regulators of phenylpropanoid metabolism in plants. Mol. Plant 8, 689–708.

    CAS  PubMed  Google Scholar 

  • Liu Y., Liu H. and Xin H. 2010 Study on the content of coumarin in different parts of Glehnia littoralis. J. Wuhan Bot. Res. 28, 114–117.

    CAS  Google Scholar 

  • Liu Y., Wang Y., Guo F., Zhan L., Mohr T., Cheng P. et al. 2017 Deep sequencing and transcriptome analyses to identify genes involved in secoiridoid biosynthesis in the Tibetan medicinal plant Swertia mussotii. Sci. Rep. 7, 43108.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Luo H., Sun C., Sun Y., Wu Q., Li Y., Song J. et al. 2011 Analysis of the transcriptome of Panax notoginseng root uncovers putative triterpene saponin-biosynthetic genes and genetic markers. BMC Genomics 12, S5.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Mi C., Ma J., Wang K. S., Zuo H. X., Wang Z., Li M. Y. et al. 2017 Imperatorin suppresses proliferation and angiogenesis of human colon cancer cell by targeting HIF-1α via the mTOR/p70S6K/4E-BP1 and MAPK pathways. J. Ethnopharmacol. 203, 27–38.

    CAS  PubMed  Google Scholar 

  • Munakata R., Olry A., Karamat F., Courdavault V., Sugiyama A., Date Y. et al. 2016 Molecular evolution of parsnip (Pastinaca sativa) membrane-bound prenyltransferases for linear and/or angular furanocoumarin biosynthesis. New Phytol. 211, 332–344.

    CAS  PubMed  Google Scholar 

  • Murata J., Roepke J., Gordon H. and De Luca V. 2008 The leaf epidermome of Catharanthus roseus reveals its biochemical specialization. Plant Cell 20, 524–542.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Patzlaff A. 2003 Characterisation of Pt MYB1, an R2R3-MYB from pine xylem. Plant Mol. Biol. 53, 597–608.

    CAS  PubMed  Google Scholar 

  • Prouse M. B. and Campbell M. M. 2013 Interactions between the R2R3-MYB transcription factor, AtMYB61, and target DNA binding sites. PLoS One 8, e65132.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Ruan Y., Le Ber P., Ng H. H. and Liu E. T. 2004 Interrogating the transcriptome. Trends Biotechnol. 22, 23–30.

    CAS  PubMed  Google Scholar 

  • Sancho R., Márquez N., Gómez-Gonzalo M., Calzado M. A., Giorgio Bettoni G., Coiras M. T. et al. 2004 Imperator in inhibits HIV-1 replication through an Sp1-dependent pathway. J. Biol. Chem. 279, 37349–37359.

    CAS  PubMed  Google Scholar 

  • Schoch G. A., Morant M., Abdulrazzak N., Asnaghi C., Goepfert S., Petersen M. et al. 2006 The meta-hydroxylation step in the phenylpropanoid pathway: a new level of complexity in the pathway and its regulation. Environ. Chem. Lett. 4, 127–136.

    CAS  Google Scholar 

  • Tamagnone L., Merida A., Parr A., Mackay S., Culianez Macia F., Roberts K. et al. 1998 The AmMYB308 and AmMYB330 transcription factors from Antirrhinum regulated phenylpropanoid and lignin biosynthesis in transgenic tobacco. Plant Cell 10, 135–154.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Wang A., Zhang P., Liu X., Liang J. and Li W. 2016a Genetic structure and diversity of Glehnia littoralis, an endangered medicinal plant in China. Biochem. Syst. Ecol. 66, 265–271.

    CAS  Google Scholar 

  • Wang H., Penmetsa R., Yuan M. and Gong L. 2012 Development and characterization of BAC-end sequence derived SSRs, and their incorporation into a new higher density genetic map for cultivated peanut (Arachis hypogaea L.) BMC Plant Biol. 12, 10.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Wang X., Cheng K., Han Y., Zhang G., Dong J., Cui Y. et al. 2016b Effects of psoralen as an anti-tumor agent in human breast cancer mcf-7/adr cells. Biol. Pharm. Bull. 39, 815–822.

    PubMed  Google Scholar 

  • Wang Z., Gerstein M. and Snyder M. 2009 RNA-Seq: a revolutionary tool for transcriptomics. Nat. Rev. Genet. 10, 57–63.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Xu Y., Wang J., Wang S., Wang J. and Chen X. 2004 Characterization of GaWRKY1, a cotton transcription factor that regulates the sesquiterpene synthase gene (+)-delta-cadinene synthase-A. Plant Physiol. 135, 507–515.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Yang L., Ding G., Lin H., Cheng H., Kong Y., Wei Y. et al. 2013 Transcriptome analysis of medicinal plant Salvia miltiorrhiza and identification of genes related to tanshinone biosynthesis. PLoS One 8, e80464.

    PubMed  PubMed Central  Google Scholar 

  • Yasumoto S., Fukushima E. O., Seki H. and Muranaka T. 2016 Novel triterpene oxidizing activity of Arabidopsis thaliana CYP716A subfamily enzymes. FEBS Lett. 590, 533–540.

    CAS  PubMed  Google Scholar 

  • Yuan Y., Song L., Li M., Liu G., Chu Y., Ma L. et al. 2012 Genetic variation and metabolic pathway intricacy govern the active compound content and quality of the Chinese medicinal plant Lonicera japonica thunb. BMC Genomics 13, 195.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Yuan Y., Wu C., Liu Y., Yang J. and Huang L. 2013 The Scutellaria baicalensis R2R3-MYB transcription factors modulates flavonoid biosynthesis by regulating GA metabolism in transgenic tobacco plants. PLoS One 8, e77275.

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The authors acknowledge the open topic of Shanghai Key Laboratory of Plant Functional Genomics and Resources (Shanghai Chenshan Botanical Garden) PFGR201703, National Natural Science Foundation of China (81903748), the Startup Foundation for Advanced Talents of Qingdao Agricultural University under Award (6631113313) and Science Technique Project of Hebei Provincial Higher Education (QN2018128).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to TING GAO.

Additional information

Corresponding editor: H. A. Ranganath

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 24384 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

SONG, J., LUO, H., XU, Z. et al. Mining genes associated with furanocoumarin biosynthesis in an endangered medicinal plant, Glehnia littoralis. J Genet 99, 11 (2020). https://doi.org/10.1007/s12041-019-1170-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s12041-019-1170-6

Keywords

Navigation