Skip to main content
SpringerLink
Log in

Enhancement of flavone levels through overexpression of chalcone isomerase in hairy root cultures of Scutellaria baicalensis

  • Short Communication
  • Published:
Functional & Integrative Genomics Aims and scope Submit manuscript

Abstract

A complementary DNA (cDNA) encoding Scutellaria baicalensis chalcone isomerase (SbCHI) was isolated using rapid amplification of cDNA ends polymerase chain reaction. After the treatment of wounding or methyl jasmonate, SbCHI transcripts were increased in S. baicalensis cell suspensions. SbCHI-overexpressed and SbCH-silenced transgenic hairy root lines were established by using an Agrobacterium rhizogenes-mediated system. SbCHI-overexpressed hairy root lines not only enhanced SbCHI gene expression but also produced more flavones (i.e., baicalin, baicalein, and wogonin) than the control hairy root line. In contrast, SbCHI-silenced hairy root lines reduced SbCHI transcripts and flavone production compared to those of the control hairy roots. In addition, the amount of wogonin in all hairy root cultures was increased compared to that of wild-type roots of S. baicalensis. Finally, this study showed the importance of CHI in flavone biosynthesis and the efficiency of metabolic engineering in S. baicalensis hairy roots.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Abbreviations

2,4-D:

2,4-Dichlorophenoxy acetic acid

4CL:

4-Coumarate:CoA ligase

BLAST:

Basic Local Alignment Search Tool

C4H:

Cinnamic acid 4-hydroxylase

CHS:

Chalcone synthase

HPLC:

High-performance liquid chromatography

LB:

Luria-Bertani

Meja:

Methyl jasmonate

MS:

Murashige and Skoog

NAA:

α-Naphthalene acetic acid

ORF:

Open reading frame

PAL:

Phenylalanine ammonia lyase

RACE-PCR:

Rapid amplification of cDNA ends polymerase chain reaction

SbCHI:

Scutellaria baicalensis chalcone isomerase

References

  • Chi YS, Cheon BS, Kim HP (2001) Effect of wogonin, a plant flavone from Scutellaria radix, on the suppression of cyclooxygenase-2 and the induction of inducible nitric oxide synthase in lipopolysaccharide-treated RAW 264.7 cells. Biochem Pharmacol 61:1195–1203

    Article  PubMed  CAS  Google Scholar 

  • Chung H, Y-m J, Shin D-H, Lee J-Y, Oh M-Y, Kim H-J, Jang KS, Jeon SJ, Son KH, Kong G (2008) Anticancer effects of wogonin in both estrogen receptor-positive and -negative human breast cancer cell lines in vitro and in nude mice xenografts. Int J Cancer 122:816–822

    Article  PubMed  CAS  Google Scholar 

  • Creelman RA, Tierney ML, Mullet JE (1992) Jasmonic acid/methyl jasmonate accumulate in wounded soybean hypocotyls and modulate wound gene expression. Proc Natl Acad Sci USA 89:4938–4941

    Article  PubMed  CAS  Google Scholar 

  • Giri A, Narasu ML (2000) Transgenic hairy roots: recent trends and applications. Biotechnol Adv 18:1–22

    Article  PubMed  CAS  Google Scholar 

  • Guillon S, Tremouillaux-Guiller J, Pati PK, Rideau M, Gantet P (2006) Hairy root research: recent scenario and exciting prospects. Curr Opin Plant Biol 9:341

    Article  PubMed  CAS  Google Scholar 

  • Han J, Ye M, Xu M, Sun JH, Wang BR, Guo D (2007) Characterization of flavonoids in the traditional Chinese herbal medicine-Huangqin by liquid chromatography coupled with electrospray ionization mass spectrometry. J Chromatogr B 848:355–362

    Article  CAS  Google Scholar 

  • Huang Y, Tsang SY, Yao X, Chen ZY (2005) Biological properties of baicalein in cardiovascular system. Curr Drug Targets Cardiovasc Haematol Disord 5:177–184

    Article  PubMed  CAS  Google Scholar 

  • Junghans H, Dalkin K, Dixon RA (1993) Stress responses In alfalfa (Medicago sativa L).15. Characterization and expression patterns of members of a subset of the chalcone synthase multigene family. Plant Mol Biol 22:239–253

    Article  PubMed  CAS  Google Scholar 

  • Kovács G, Kuzovkina IN, Szoke É, Kursinszki L (2004) HPLC determination of flavonoids in hairy-root cultures of Scutellaria baicalensis Georgi. Chromatographia 60:S81–S85

    Article  Google Scholar 

  • Kuzovkina IN, Guseva AV, Alterman IE, Karnachuk RA (2001) Flavonoid production in transformed Scutellaria baicalensis roots and ways of its regulation. Russ J Plant Physiol 48:448–452

    Article  CAS  Google Scholar 

  • Kuzovkina IN, Guseva AV, Kovács D, Szöke É, Vdovitchenko MY (2005) Flavones in genetically transformed Scutellaria baicalensis roots and induction of their synthesis by elicitation with methyl jasmonate. Russ J Plant Physiol 52:77–82

    Article  CAS  Google Scholar 

  • Lee H, Kim YO, Kim H, Kim SY, Noh HS, Kang SS, Cho GJ, Choi WS, Suk K (2003) Flavonoid wogonin from medicinal herb is neuroprotective by inhibiting inflammatory activation of microglia. FASEB J 17:1943–1944. doi:10.1096/fj.03-0057fje

    PubMed  CAS  Google Scholar 

  • Lee D-H, Kim C, Zhang L, Lee YJ (2008) Role of p53, PUMA, and Bax in wogonin-induced apoptosis in human cancer cells. Biochem Pharmacol 75:2020–2033

    Article  PubMed  CAS  Google Scholar 

  • Li H, Murch S, Saxena P (2000) Thidiazuron-induced de novo shoot organogenesis on seedlings, etiolated hypocotyls and stem segments of Huang-qin. Plant Cell Tissue Organ Cult 62:169–173

    Article  CAS  Google Scholar 

  • Li F-X, Jin Z-P, Qu W, Zhao D-X, Ma F (2006a) Cloning of cDNA encoding the Saussurea medusa chalcone isomerase and its expression in transgenic tobacco. Plant Physiol Biochem 44:455–461

    Article  PubMed  CAS  Google Scholar 

  • Li F-X, Jin Z-P, Zhao D-X, Cheng L-Q, Fu C-X, Ma F (2006b) Overexpression of the Saussurea medusa chalcone isomerase gene in S. involucrata hairy root cultures enhances their biosynthesis of apigenin. Phytochemistry 67:553–560

    Article  PubMed  CAS  Google Scholar 

  • Liu S, Zhang J, Li D, Liu W, Luo X, Zhang R, Li L, Zhao J (2007) Anticancer activity and quantitative analysis of flavone of Cirsium japonicum DC. Nat Prod Res formerly Nat Prod Lett 21:915–922

    Google Scholar 

  • Li-Weber M (2009) New therapeutic aspects of flavones: the anticancer properties of Scutellaria and its main active constituents Wogonin, Baicalein and Baicalin. Cancer Treat Rev 35:57–68

    Article  PubMed  CAS  Google Scholar 

  • Lu N, Gao Y, Ling Y, Chen Y, Yang Y, Gu H-Y, Qi Q, Liu W, Wang X-T, You Q-D, Guo Q-L (2008) Wogonin suppresses tumor growth in vivo and VEGF-induced angiogenesis through inhibiting tyrosine phosphorylation of VEGFR2. Life Sci 82:956–963

    Article  PubMed  CAS  Google Scholar 

  • Murch SJ, Rupasinghe HPV, Goodenowe D, Saxena PK (2004) A metabolomic analysis of medicinal diversity in Huang-qin (Scutellaria baicalensis Georgi) genotypes: discovery of novel compounds. Plant Cell Rep 23:419–425

    Article  PubMed  CAS  Google Scholar 

  • Nishikawa K, Furukawa H, Fujioka T, Fujii H, Mihashi K, Shimomura K, Ishimaru K (1999) Flavone production in transformed root cultures of Scutellaria baicalensis Georgi. Phytochemistry 52:885–890

    Article  CAS  Google Scholar 

  • Parajuli P, Joshee N, Rimando AM, Mittal S, Yadav AK (2009) In vitro antitumor mechanisms of various Scutellaria extracts and constituent flavonoids. Planta Med 75:41–48

    Article  PubMed  CAS  Google Scholar 

  • Richard S, Lapointe G, Rutledge RG, Seguin A (2000) Induction of chalcone synthase expression in white spruce by wounding and jasmonate. Plant Cell Physiol 41:982–987. doi:10.1093/pcp/pcd017

    Article  PubMed  CAS  Google Scholar 

  • Shieh DE, Liu LT, Lin CC (2000) Antioxidant and free radical scavenging effects of baicalein, baicalin and wogonin. Anticancer Res 20:2861–2865

    PubMed  CAS  Google Scholar 

  • Shimada N, Aoki T, Sato S, Nakamura Y, Tabata S, S-i A (2003) A cluster of genes encodes the two types of chalcone isomerase involved in the biosynthesis of general flavonoids and legume-specific 5-deoxy(iso)flavonoids in Lotus japonicus. Plant Physiol 131:941–951. doi:10.1104/pp.004820

    Article  PubMed  CAS  Google Scholar 

  • Wang W, Guo Q-L, You Q-D, Zhang K, Yang Y, Yu J, Liu W, Zhao L, Gu H-Y, Hu Y, Tan Z, Wang X-T (2006) The anticancer activities of wogonin in murine sarcoma S180 both in vitro and in vivo. Biol Pharm Bull 29:1132–1137

    Article  PubMed  CAS  Google Scholar 

  • Wasson AP, Pellerone FI, Mathesius U (2006) Silencing the flavonoid pathway in Medicago truncatula inhibits root nodule formation and prevents auxin transport regulation by rhizobia. Plant Cell 18:1617–1629. doi:10.1105/tpc.105.038232

    Article  PubMed  CAS  Google Scholar 

  • Xu H, Park NI, Li X, Kim YK, Lee SY, Park SU (2010) Molecular cloning and characterization of phenylalanine ammonia-lyase, cinnamate 4-hydroxylase and genes involved in flavone biosynthesis in Scutellaria baicalensis. Bioresour Technol 101:9715–9722

    Article  PubMed  CAS  Google Scholar 

  • Zhang H-C, Liu J-M, Lu H-Y, Gao S-L (2009) Enhanced flavonoid production in hairy root cultures of Glycyrrhiza uralensis Fisch by combining the over-expression of chalcone isomerase gene with the elicitation treatment. Plant Cell Rep 28:1205–1213

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The presented data are results of a study on the "Human Resource Development Center for Economic Region Leading Industry" Project, supported by the Ministry of Education, Science & Technology (MEST) and the National Research Foundation of Korea (NRF).

Author information

Authors and Affiliations

  1. Department of Crop Science, College of Agriculture and Life Sciences, Chungnam National University, 220 Gung-dong, Yuseong-gu, Daejeon, 305–764, South Korea

    Nam Il Park, Hui Xu, Xiaohua Li & Sang Un Park

  2. Department of Bio-Environmental Chemistry, College of Agriculture and Life Sciences, Chungnam National University, 220 Gung-dong, Yuseong-gu, Daejeon, 305–764, South Korea

    Sun-Ju Kim

Authors
  1. Nam Il Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hui Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiaohua Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sun-Ju Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sang Un Park
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sang Un Park.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

The online version of this article contains supplementary material, which is available to authorized users (PDF 848 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Park, N.I., Xu, H., Li, X. et al. Enhancement of flavone levels through overexpression of chalcone isomerase in hairy root cultures of Scutellaria baicalensis . Funct Integr Genomics 11, 491–496 (2011). https://doi.org/10.1007/s10142-011-0229-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10142-011-0229-0

Keywords

Search

Navigation