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Characterization of the Scutellaria barbata glycosyltransferase gene and its promoter

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Abstract

The conversion of flavonoid aglycones to their glycosides by plant glycosyltransferases may affect a wide range of outcomes, including stability, solubility and bioavailability. Scutellaria barbata, rich in flavonoid glycosides, is widely used as a traditional Chinese herbal medicine. In this study, a flavonoid glycosyltransferase cDNA (SbUGT) and its promoter from S. barbata were cloned and characterized as a flavonoid glycosyltransferase using whole-cell biotransformation. Fragments of different lengths of the 5′-flanking region of the SbUGT gene were fused to the β-glucuronidase (GUS) gene and analyzed with transgenic Arabidopsis plants using histochemical and fluorometric assays. GUS activity in transgenic plants carrying the SbP-850U construct (−850 to +86 relative to the transcription start site) displayed the highest level and was enhanced by salt and methyl jasmonate, similar to the expression patterns of the endogenous SbUGT. GUS activity disappeared when the promoter was deleted to −98, and deletion analyses indicated the existence of positive and negative regulatory element(s). Unexpectedly, plants carrying the construct SbP-102U (−102 to +86) exhibited strong GUS activity exclusively in the roots. Our experiments revealed that the specific expression is mediated by different promoter regions and the unique region driving root-preferred expression can be used as a root-specific promoter.

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Abbreviations

ABA:

Abscisic acid

CTAB:

Hexadecyltrimethylammonium bromide

GUS:

β-Glucuronidase

MU:

4-Methylumbelliferone

MeJA:

Methyl jasmonate

SA:

Salicylic acid

SPE:

Strong positive element

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Acknowledgments

We thank Michel Delseny at the Centre National De La Recherche Scientifique (CNRS) and C.Y. Lee at ITRI for comments on the manuscript. We also thank Ching Li and Hui-Yen Wu at ITRI for assistance with LC/MS analysis. This study was supported by the Ministry of Economic Affairs (98-EC-17-A-02-04-0317) and the National Science Council (NSC 98-2313-B-007-001-MY3), Republic of China.

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Authors and Affiliations

  1. Department of Life Science, Institute of Bioinformatics and Structural Biology, National Tsing Hua University, No. 101, Sec. 2, Kuang Fu Rd., Hsinchu, 30013, Taiwan, ROC

    Shu-Jiau Chiou, Wen-Yu Liu & Tsai-Yun Lin

  2. Biomedical Technology and Device Research Labs, Industrial Technology Research Institute, Hsinchu, Taiwan, ROC

    Shu-Jiau Chiou & Cheng-Li Fang

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  1. Shu-Jiau Chiou
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  2. Wen-Yu Liu
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Corresponding author

Correspondence to Tsai-Yun Lin.

Additional information

The nucleotide sequences reported in this study were deposited in the GenBank database with accession numbers GU339042 and GU339043.

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Chiou, SJ., Liu, WY., Fang, CL. et al. Characterization of the Scutellaria barbata glycosyltransferase gene and its promoter. Planta 232, 963–974 (2010). https://doi.org/10.1007/s00425-010-1229-3

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  • DOI: https://doi.org/10.1007/s00425-010-1229-3

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