Skip to main content
SpringerLink
Log in

Characterization of a chalcone synthase (CHS) flower-specific promoter from Lilium orential ‘Sorbonne’

  • Original Paper
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

The first enzyme in the flavonoid pathway, chalcone synthase, is encoded by a gene (CHS) whose expression is normally under developmental control. In our previous studies, an 896-bp promoter region of a flower-specific CHS gene was isolated from Lilium orential ‘Sorbonne’, and designated as PLoCHS. Here, the PLoCHS promoter was fused to the β-glucuronidase (GUS) gene to characterize its spatial and temporal expression in Petunia hybrida ‘Dreams Midnight’ using an Agrobacterium-mediated leaf disc transformation method. Our results demonstrated that GUS expression was present in flowers, but reduced or absent in the other tissues (leaf and stem) examined. In petals, GUS activity reached its peak at flower developmental stage 4, and decreased at later stages. Deletion analysis indicated that even a 307-bp fragment of the PLoCHS promoter could still direct flower-specific expression. Further deletion of the region from −261 to −72 bp resulted in weak expression in different organs, including flowers, leaves and stems. This evidence combined with prediction of cis-acting elements in the PLoCHS promoter suggests that the TACPyAT box located in this promoter plays a key role in the regulation of organ-specific expression.

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
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

CHS:

Chalcone synthase

GUS:

β-Glucuronidase

DFR:

Dihydroflavonol 4-reductase

ACO:

1-Aminocyclopropane-1-carboxylate oxidase

CaMV 35S:

Cauliflower mosaic virus 35S promoter

NOS:

Nopaline synthase

MU:

4-Methyl umbelliferone

MUG:

4-Methyl umbelliferyl glucuronide

MS:

Murashige and Skoog’s medium

BA:

6-Benzyladenine

NAA:

α-Naphthaleneacetic acid

TSS:

Transcription start site

X-Gluc:

5-Bromo-4-chloro-3-indolyl-b-d-glucuronic acid

References

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  • Chiou CY, Wu K, Yeh KW (2008) Characterization and promoter activity of chromoplast specific carotenoid associated gene (CHRC) from Oncidium Gower Ramsey. Biotechnol Lett 30:1861–1866

    Article  PubMed  CAS  Google Scholar 

  • Cone KC, Cocciolone SM, Burr FA, Burr B (1993) Maize anthocyanin regulatory gene pl is a duplicate of c1 that functions in the plant. Plant Cell 5:1795–1805

    Article  PubMed  CAS  Google Scholar 

  • Faktor O, Kooter JM, Dixon RA, Lamb CJ (1996) Functional dissection of a bean chalcone synthase gene promoter in transgenic tobacco plants reveals sequence motifs essential for floral expression. Plant Mol Biol 32:849–859

    Article  PubMed  CAS  Google Scholar 

  • Farzad M, Griesbach R, Hammond J, Weiss MR, Elmendorf HG (2003) Differential expression of three key anthocyanin biosynthetic genes in a color-changing flower, Viola cornuta cv. Yesterday, Today and Tomorrow. Plant Sci 165:1333–1342

    Article  CAS  Google Scholar 

  • Filichkin SA, Leonard JM, Monteros A, Liu PP, Nonogaki H (2004) A novel endo-betamannanase gene in tomato LeMAN5 is associated with anther and pollen development. Plant Physiol 134:1080–1087

    Article  PubMed  CAS  Google Scholar 

  • Han YY, Ming F, Wang JW, Ye MM, Shen DL (2005) Molecular characterization and functional analysis of a novel chalcone synthase gene from Phalaenopsis Orchid in transgenic tobacco. Plant Mol Biol Rep 23:193a–193m

    Google Scholar 

  • Han YY, Ming F, Wang W, Wang JW, Ye MM, Shen DL (2006) Molecular evolution and functional specialization of chalcone synthase superfamily from Phalaenopsis Orchid. Genetica 128:429–438

    Article  PubMed  CAS  Google Scholar 

  • Helariutta Y, Elomaa P, Kotilainen M, Griesbach RJ, SchrÖder J, Teeri TH (2004) Chalcone synthase-like genes active during corolla development are differentially expressed and encode enzymes with different catalytic properties in Gerbera hybrida (Asteraceae). Plant Mol Biol 28:47–60

    Article  Google Scholar 

  • Himi E, Noda K (2004) Isolation and location of three homoeologous dihydroflavonol-4-reductase (DFR) genes of wheat and their tissue-dependent expression. J Exp Bot 55:365–375

    Article  PubMed  CAS  Google Scholar 

  • Himi E, Nisar A, Noda K (2005) Colour genes (R and Rc) for grain and coleoptiles upregulate flavonoid biosynthesis genes in wheat. Genome 48:747–754

    Article  PubMed  CAS  Google Scholar 

  • Hraška M, Rakouský S, Čurn V (2008) Tracking of the CaMV-35S promoter performance in GFP transgenic tobacco, with a special emphasis on flowers and reproductive organs, confirmed its predominant activity in vascular tissues. Plant Cell Tissue Organ Cult 94:239–251

    Article  Google Scholar 

  • Huits HSM, Gerats AGM, Kreike MM, Mol JNM, Koes RE (1994) Genetic control of dihydroflavonol 4-reductase gene expression in Petunia hybrida. Plant J 6:295–310

    Article  PubMed  CAS  Google Scholar 

  • Ito M, Ichinose Y, Kato H, Shiraishi T, Yamada T (1997) Molecular evolution and functional relevance of the chalcone synthase genes of pea. Mol Gen Genet 255:28–37

    Article  PubMed  CAS  Google Scholar 

  • Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405

    Article  CAS  Google Scholar 

  • Kobayashi H, Oikawa Y, Koiwa H, Yamamura S (1998) Flower-specific gene expression directed by the promoter of a chalcone synthase gene from Gentiana triflora in Petunia hybrida. Plant Sci 131:173–180

    Article  CAS  Google Scholar 

  • Koduri PKH, Gordon GS, Barker EI, Colpitts CC, Ashton NW, Suh DY (2010) Genome-wide analysis of the chalcone synthase superfamily genes of Physcomitrella patens. Plant Mol Biol 72:247–263

    Article  PubMed  CAS  Google Scholar 

  • Koes RE, Spelt CE, Reif HJ, Mol JNM (1989) The chalcone synthase multigene family of Petunia hybrid (V30):differential, light-regulated expression during flower development and UV light induction. Plant Mol Biol 12:213–225

    Article  CAS  Google Scholar 

  • Liew CF, Loh CS, Gho CJ, Lim SH (1998) The isolation, molecular characterization and expression of dihydroflavonol 4-reductase cDNA in the orchid, Bromheadia finlaysoniana. Plant Sci 135:161–169

    Article  CAS  Google Scholar 

  • Ludwig SR, Habera LF, Dellaporta ST, Wessler SR (1989) Lc, a member of the maze R gene family responsible for tissue specific anthocyanin production, encodes a protein similar to transcriptional activators and contains the myc-homology region. Proc Natl Acad Sci USA 86:7092–7096

    Article  PubMed  CAS  Google Scholar 

  • McKhann HI, Hirsch AM (1994) Isolation of chalcone synthase and chalcone isomerase cDNAs from alfalfa (Medicago sativa L.): Highest transcript levels occur in young roots and root tips. Plant Mol Biol 24:767–777

    Article  PubMed  CAS  Google Scholar 

  • Meng XC, Xing T, Wang XJ (2004) The role of light in the regulation of anthocyanin accumulation in Gerbera hybrida. Plant Growth Regul 44:243–250

    Article  CAS  Google Scholar 

  • Mitsuda N, Takeyasu K, Sato MH (2001) Pollen-specific regulation of vacuolar H+-PPase expression by multiple cis-acting elements. Plant Mol Biol 46:185–192

    Article  PubMed  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–479

    Article  CAS  Google Scholar 

  • Nakatsuka A, Izumi Y, Yamagishi M (2003) Spatial and temporal expression of chalcone synthase and dihydroflavonol 4-reductase genes in the Asiatic hybrid lily. Plant Sci 165:759–767

    Article  CAS  Google Scholar 

  • Rogers HJ, Bate N, Combe J, Sullivan J, Sweetman J, Swan C, Lonsdale DM, Twell D (2001) Functional analysis of cis-regulatory elements within the promoter of the tobacco late pollen gene g10. Plant Mol Biol 45:577–585

    Article  PubMed  CAS  Google Scholar 

  • Saito R, Fukuta N, Ohmiya A, Itoh Y, Ozeki Y, Kuchitsu K, Nakayama M (2006) Regulation of anthocyanin biosynthesis involved in the formation of marginal picotee petals in Petunia. Plant Sci 70:828–834

    Article  Google Scholar 

  • Tai TH, Tanksley SD (1990) A rapid and inexpensive method for isolation of total DNA from dehydrated plant tissue. Plant Mol Biol Rep 8:229–303

    Article  Google Scholar 

  • Tanaka Y, Katsumoto Y, Brugliera F, Mason J (2005) Genetic engineering in floriculture. Plant Cell Tiss Org Cult 80:1–24

    Article  CAS  Google Scholar 

  • Uimari A, Strommer J (1997) Myb26: a MYB-like protein of pea flowers with affinity for promoters of phenylpropanoid genes. Plant J 12:1273–1284

    Article  PubMed  CAS  Google Scholar 

  • Van der Meer IM (2006) Agrobacterium-mediated transformation of petunia leaf discs. Plant Cell Cult Protoc doi:10.1385/1-59259-959-1:265

  • Van der Meer IM, Spelt CE, Mol JNM, Stuitje AR (1990) Promoter analysis of the chalcone synthase (CHSA) gene of Petunia hybrida: a 67 bp promoter region directs flower-specific expression. Plant Mol Biol 15:95–109

    Article  PubMed  Google Scholar 

  • Van der Meer IM, Brouwer M, Spelt CE, Mol JNM, Stuitje AR (1992) The TACPyAT repeats in the chalcone synthase promoter of Petunia hybrida act as a dominant negative cis-acting module in the control of organ-specific expression. Plant J 2:525–535

    Article  PubMed  Google Scholar 

  • Yang L (2006) Cloning and functional analysis of CHS and its promoter in Lilium. Dissertation, Northwest A&F University

  • Yang J, Gu HY (2006) Duplication and divergent evolution of the CHS and CHS-like genes in the chalcone synthase (CHS) superfamily. Chin Sci Bull 51:505–509

    Article  CAS  Google Scholar 

  • Yi JX, Derynck MR, Chen L, Dhaubhadel S (2010) Differential expression of CHS7 and CHS8 genes in soybean. Planta 231:741–753

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We would like to thank Li Li for helpful suggestions. This work was supported by the National Natural Science Foundation of China (Grant no: 30871734).

Author information

Authors and Affiliations

  1. College of Forestry, Northwest A&F University, Yangling, 712100, Shaanxi, People’s Republic of China

    Yali Liu & Yi Xin

  2. College of Horticulture, Northwest A&F University, Yangling, 712100, Shaanxi, People’s Republic of China

    Qian Lou, Weirong Xu & Yuejin Wang

  3. Key Laboratory of Horticulture Plant Germplasm Utilization in Northwest China of Ministry of Agriculture, Yangling, 712100, Shaanxi, People’s Republic of China

    Yali Liu, Qian Lou, Weirong Xu, Yi Xin & Yuejin Wang

  4. State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A&F University, Yangling, 712100, Shaanxi, People’s Republic of China

    Yali Liu, Qian Lou, Weirong Xu, Yi Xin & Yuejin Wang

  5. USDA-ARS, Appalachian Fruit Research Station, 2217 Wiltshire Road, Kearneysville, WV, 25430, USA

    Carole Bassett

Authors
  1. Yali Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Qian Lou
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Weirong Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yi Xin
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Carole Bassett
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yuejin Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yali Liu.

Additional information

Communicated by W. Harwood.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Liu, Y., Lou, Q., Xu, W. et al. Characterization of a chalcone synthase (CHS) flower-specific promoter from Lilium orential ‘Sorbonne’. Plant Cell Rep 30, 2187–2194 (2011). https://doi.org/10.1007/s00299-011-1124-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-011-1124-9

Keywords

Search

Navigation