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Arabidopsis pab1, a mutant with reduced anthocyanins in immature seeds from banyuls, harbors a mutation in the MATE transporter FFT

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Abstract

Forward genetics approaches have helped elucidate the anthocyanin biosynthetic pathway in plants. Here, we used the Arabidopsis banyuls (ban) mutant, which accumulates anthocyanins, instead of colorless proanthocyanidin precursors, in immature seeds. In contrast to standard screens for mutants lacking anthocyanins in leaves/stems, we mutagenized ban plants and screened for mutants showing differences in pigmentation of immature seeds. The pale banyuls1 (pab1) mutation caused reduced anthocyanin pigmentation in immature seeds compared with ban. Immature pab1 ban seeds contained less anthocyanins and flavonols than ban, but showed normal expression of anthocyanin biosynthetic genes. In contrast to pab1, introduction of a flavonol-less mutation into ban did not produce paler immature seeds. Map-based cloning showed that two independent pab1 alleles disrupted the MATE-type transporter gene FFT/DTX35. Complementation of pab1 with FFT confirmed that mutation in FFT causes the pab1 phenotype. During development, FFT promoter activity was detected in the seed-coat layers that accumulate flavonoids. Anthocyanins accumulate in the vacuole and FFT fused to GFP mainly localized in the vacuolar membrane. Heterologous expression of grapevine MATE-type anthocyanin transporter gene partially complemented the pab1 phenotype. These results suggest that FFT acts at the vacuolar membrane in anthocyanin accumulation in the Arabidopsis seed coat, and that our screening strategy can reveal anthocyanin-related genes that have not been found by standard screening.

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Acknowledgments

We thank Dr. Martine Devic for providing ban-1, Dr. Agnès Ageorges for providing pGEM-AM3, and the Arabidopsis Biological Resource Center and RIKEN-BRC for providing T-DNA insertion mutant lines. We are grateful to members of the Ion Beam Mutagenesis Research Group at JAEA for kind help and valuable discussions.

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  1. Issay Narumi

    Present address: Faculty of Life Sciences, Toyo University, Gunma, 374-0193, Japan

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  1. Ion Beam Mutagenesis Research Group, Medical and Biotechnological Application Unit, Quantum Beam Science Center, Japan Atomic Energy Agency (JAEA), 1233 Watanuki, Takasaki, 370-1292, Japan

    Satoshi Kitamura, Yutaka Oono & Issay Narumi

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  1. Satoshi Kitamura
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Correspondence to Satoshi Kitamura.

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Kitamura, S., Oono, Y. & Narumi, I. Arabidopsis pab1, a mutant with reduced anthocyanins in immature seeds from banyuls, harbors a mutation in the MATE transporter FFT . Plant Mol Biol 90, 7–18 (2016). https://doi.org/10.1007/s11103-015-0389-8

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