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Isolation and functional characterization of a R2R3-MYB regulator of the anthocyanin biosynthetic pathway from Epimedium sagittatum

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A R2R3-MYB transcription factor EsAN2 was isolated from Epimedium sagittatum and functionally characterized to regulate the anthocyanin biosynthetic pathway.

Abstract

Epimedium plants are used widely both as traditional Chinese medicinal herbs and ornamental perennials. Anthocyanins, acting as major contributors to plant color diversity, their biosynthesis are regulated by a series of transcription factors, including MYB, bHLH and WD40 protein. Previously, a MYB transcription factor involved in regulation of the anthocyanin pathway from Epimedium sagittatum, EsMYBA1 has been isolated, but was found to be expressed mostly in leaves. In this research, another MYB transcription factor, designated as EsAN2, was isolated from flowers by the screening of E. sagittatum EST database. Preferential expression of EsAN2 in flowers and flower buds was found. Ectopic expression of EsAN2 in tobacco significantly enhanced the anthocyanin biosynthesis and accumulation, both in leaves and flowers. Most structural genes of the anthocyanin biosynthetic pathway were strongly upregulated, as well as two bHLH regulators (NtAn1a and NtAn1b) in old leaves of tobacco overexpressing EsAN2, compared to the control plants. While only three structural genes, chalcone synthase (CHS), chalcone isomerase (CHI) and anthocyanidin synthase (ANS), were upregulated by EsAN2 ectopic expression in tobacco flowers. Yeast two-hybrid assay showed that EsAN2 was capable of interacting with four bHLH regulators of the anthocyanin biosynthetic pathway. These results suggest that EsAN2 is involved in regulation of the anthocyanin biosynthesis in Epimedium flowers. Identification and characterization of EsAN2 provide insight into the coloration of Epimedium flowers and a potential candidate gene for metabolic engineering of flavonoids in the future.

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Author contribution statement

WJH, and YW designed the research and wrote the paper. WJH and ABMK performed most of the experiments. HYL, LWD and CJZ participated in the experiments. All authors read and approved the manuscript.

Acknowledgments

This study was supported by grants from the National Natural Science Foundation of China (No. 31200225, 31270340). We thank Dr. Xianbao Deng for critical editing.

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    Authors and Affiliations

    1. Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture, Wuhan Botanical Garden, Chinese Academy of Sciences, 430074, Wuhan, Hubei, China

      Wenjun Huang, A. B. M. Khaldun, Haiyan Lv, Liuwen Du & Ying Wang

    2. University of the Chinese Academy of Sciences, 100039, Beijing, China

      Liuwen Du

    3. Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute of the Chinese Academy of Agriculture Sciences, 430062, Wuhan, Hubei, China

      Chanjuan Zhang

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    1. Wenjun Huang
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    Correspondence to Ying Wang.

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    Communicated by Z.-Y. Wang.

    W. Huang and A. B. M. Khaldun contributed equally to this work.

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    Huang, W., Khaldun, A.B.M., Lv, H. et al. Isolation and functional characterization of a R2R3-MYB regulator of the anthocyanin biosynthetic pathway from Epimedium sagittatum . Plant Cell Rep 35, 883–894 (2016). https://doi.org/10.1007/s00299-015-1929-z

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