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Functional characterization of key structural genes in rice flavonoid biosynthesis

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Abstract

Rice is a model system for monocot but the molecular features of rice flavonoid biosynthesis have not been extensively characterized. Rice structural gene homologs encoding chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), flavonoid 3′-hydroxylase (F3′H), dihydroflavonol 4-reductase (DFR), and anthocyanidin synthase (ANS) were identified by homology searches. Unique differential expression of OsF3H, OsDFR, and OsANS1 controlled by the Pl w locus, which contains the R/B-type regulatory genes OSB1 and OSB2, was demonstrated during light-induced anthocyanin accumulation in T65-Plw seedlings. Previously, F3H genes were often considered as early genes co-regulated with CHS and CHI genes in other plants. In selected non-pigmented rice lines, OSB2 is not expressed following illumination while their expressed OSB1sequences all contain the same nucleotide change leading to the T64 M substitution within the conserved N-terminal interacting domain. Furthermore, the biochemical roles of the expressed rice structural genes (OsCHS1, OsCHI, OsF3H, and OsF3′H) were established in planta for the first time by complementation in the appropriate Arabidopsis transparent testa mutants. Using yeast two-hybrid analysis, OsCHS1 was demonstrated to interact physically with OsF3H, OsF3′H, OsDFR, and OsANS1, suggesting the existence of a macromolecular complex for anthocyanin biosynthesis in rice. Finally, flavones were identified as the major flavonoid class in the non-pigmented T65 seedlings in which the single-copy OsF3H gene was not expressed. Competition between flavone and anthocyanin pathways was evidenced by the significant reduction of tricin accumulation in the T65-Plw seedlings.

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Abbreviations

AD:

Activation domain

ANS:

Anthocyanidin synthase

BD:

Binding domain

bHLH:

Basic helix-loop-helix

CHS:

Chalcone synthase

CHI:

Chalcone isomerase

DFR:

Dihydroflavonol 4-reductase

F3H:

Flavanone 3-hydroxylase

F3′H:

Flavonoid 3′-hydroxylase

FGT:

Flavonoid 3-O-glucosyltransferase

ONPG:

o-Nitrophenyl-β-d-galactopyranoside

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Acknowledgments

We thank Prof S Iida of National Institute for Basic Biology (Okazaki, Japan) for the OsDFR cDNA clone and the National Institute of Agrobiological Sciences (Tsukuba, Japan) for the OsCHS1, OsCHI, and OsF3′H clones. This work was supported by the Research Grants Council of the Hong Kong Special Administrative Region, China (grant no. HKU 7337/04M).

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Authors and Affiliations

  1. School of Biological Sciences, The University of Hong Kong, Pokfulam Road, Hong Kong, China

    Chun Hat Shih, Hung Chu, Lee Kwan Tang, Mingfu Wang & Clive Lo

  2. Department of Chemistry, The University of Hong Kong, Pokfulam Road, Hong Kong, China

    Ivan K. Chu

  3. Research Institute for Bioresources, Okayama University, Okayama, Japan

    Wataru Sakamoto & Masahiko Maekawa

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  1. Chun Hat Shih
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  2. Hung Chu
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  3. Lee Kwan Tang
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  4. Wataru Sakamoto
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  5. Masahiko Maekawa
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  6. Ivan K. Chu
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  7. Mingfu Wang
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  8. Clive Lo
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Correspondence to Clive Lo.

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Shih, C.H., Chu, H., Tang, L.K. et al. Functional characterization of key structural genes in rice flavonoid biosynthesis. Planta 228, 1043–1054 (2008). https://doi.org/10.1007/s00425-008-0806-1

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  • DOI: https://doi.org/10.1007/s00425-008-0806-1

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