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The soybean R2R3 MYB transcription factor GmMYB100 negatively regulates plant flavonoid biosynthesis

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Abstract

Soybean flavonoids, a group of important signaling molecules in plant-environment interaction, ubiquitously exist in soybean and are tightly regulated by many genes. Here we reported that GmMYB100, a gene encoding a R2R3 MYB transcription factor, is involved in soybean flavonoid biosynthesis. GmMYB100 is mainly expressed in flowers, leaves and immature embryo, and its level is decreased after pod ripening. Subcellular localization assay indicates that GmMYB100 is a nuclear protein. GmMYB100 has transactivation ability revealed by a yeast functional assay; whereas bioinformatic analysis suggests that GmMYB100 has a negative function in flavonoid biosynthesis. GmMYB100-overexpression represses the transcript levels of flavonoid-related genes in transgenic soybean hairy roots and Arabidopsis, and inhibits isoflavonoid (soybean) and flavonol (Arabidopsis) production in transgenic plants. Furthermore, the transcript levels of six flavonoid-related genes and flavonoid (isoflavonoid and flavone aglycones) accumulation are elevated in the GmMYB100-RNAi transgenic hairy roots. We also demonstrate that GmMYB100 protein depresses the promoter activities of soybean chalcone synthase and chalcone isomerase. These findings indicate that GmMYB100 is a negative regulator in soybean flavonoid biosynthesis pathway.

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Authors and Affiliations

  1. Key Laboratory of Urban Agriculture (South) Ministry of Agriculture, School of Agriculture and Biology, Shanghai Jiao Tong University, No. 800 Dongchuan Rd., Shanghai, 200240, People’s Republic of China

    Junhui Yan, Biao Wang, Yunpeng Zhong, Luming Yao, Lin**g Cheng & Tianlong Wu

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  1. Junhui Yan
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  2. Biao Wang
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  3. Yunpeng Zhong
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  4. Luming Yao
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  5. Lin**g Cheng
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  6. Tianlong Wu
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Correspondence to Biao Wang.

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Fig. S1

The expression pattern of the four genes (TF54, TF62, TF88 and TF100) under different stresses. Fold, the ratio of the gene expression level in treated seedlings compared with the control ones. JA-L and JA-R, the leaves (JA-L) and roots (JA-R) from the seedlings treated by MeJA; SA-R, Nod-R and Suc-R represent the roots from the roots treated by SA, Bradyrhizobium japonicum and sucrose, respectively. The expression levels were detected by real-time PCR and the housekee** genes are GmACTIN (V00450.1), CYB2 (TC224926), and EFL1B (TC203623) (TIFF 304 kb)

Fig. S2

SDS-PAGE analysis of the expression of GmMYB100 protein in E. coli cells harboring the pEASY-E1-GmMYB100 plasmid after induction of 0.5 mM IPTG for 0–6 h at 28 °C. M, the molecular mass protein standards (TIFF 673 kb)

Fig. S3

The expression levels of GmMYB100 in transgenic soybean hairy roots. The expression levels were detected by real-time PCR in different transgenic hairy roots lines, and the values are mean ± SE. Asterisks indicate a significant difference (P < 0.05, Student’s t test) between the transgenic soybean hairy roots. Vector Control, hairy roots transformed with pHB-YFP; GmMYB100-OE, GmMYB100-overexpression of transgenic soybean hairy roots; GmMYB100-RNAi, GmMYB100 silence of transgenic soybean hairy roots (TIFF 226 kb)

Fig. S4

Analysis of GmMYB100-overexpression transgenic Arabidopsis seedlings by PCR and RT-PCR. P, pHB-GmMYB100-OE vector; Control, the pHB-YFP transgenic Arabidopsis (T3 lines); 1–9, the GmMYB100 overexpressed transgenic Arabidopsis (T3 lines); the primers GmMYB100-2F/2R were used in RT-PCR and PCR for GmMYB100; the primers GmACTIN-F/R were used in PCR for GmACTIN (TIFF 274 kb)

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Yan, J., Wang, B., Zhong, Y. et al. The soybean R2R3 MYB transcription factor GmMYB100 negatively regulates plant flavonoid biosynthesis. Plant Mol Biol 89, 35–48 (2015). https://doi.org/10.1007/s11103-015-0349-3

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