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Molecular Cloning and Expression Analysis of an ANS Gene Encoding Anthocyanidin Synthase from Purple-Fleshed Sweet Potato [Ipomoea batatas (L.) Lam]

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Abstract

Anthocyanidin synthase (ANS), a 2-oxoglutarate (2OG) iron-dependent oxygenase, catalyzes the penultimate step in the biosynthesis of anthocyanin. This reaction is responsible for the formation of the colored anthocyanidins from the colorless leucoanthocyanidins. A full-length cDNA was isolated from purple-fleshed sweet potato (Ipomoea batatas (L.) Lam) cv. Yamakawamurasaki, designated IbANS, containing a 1,086-bp open reading frame encoding a 362-amino-acid polypeptide. Multiple alignments revealed that the deduced IbANS protein had high identity to ANS proteins of other plants such as Ipomoea nil (90.8% identities), Ipomoea purpurea (91.4% identities), and Brassica juncea (72.7% identities). Structural analysis showed that the IbANS protein might belong to the 2OG and Fe(II)-dependent oxygenase, containing three binding sites of 2OG (H236, D238, and H292) and three binding sites of Fe(II) (Y221, R302, and S304). Phylogenetic tree analysis revealed that IbANS shared the close relationships with I. nil and I. purpurea. Southern blotting showed that there were two copies of the IbANS gene in this genome. Real-time quantitative polymerase chain reaction revealed that expression of the IbANS gene was highest in storage roots and lowest in leaves. IbANS was expressed most abundantly during the formation of storage roots. In five cultivars of sweet potato, IbANS expression was strongly associated with anthocyanin accumulation, suggesting that ANS gene expression was associated with activation of anthocyanin biosynthesis.

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Abbreviations

ORF:

open reading frame

RACE:

rapid amplification of cDNA ends

UTR:

untranslated region

RT-PCR:

reverse transcription polymerase chain reaction

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Acknowledgements

This work was funded by the China National Natural Science Foundation (30671320) and Guangdong Natural Science Foundation (05005945).

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Authors and Affiliations

  1. Guangdong Key Laboratory of Biotechnology for Plant Development, School of Life Sciences, South China Normal University, Guangzhou, 510631, People’s Republic of China

    Wei Zhou, Chengtao Huang, Yifu Gong, Qili Feng & Feng Gao

  2. Faculty of Life Science and Biotechnology, Ningbo University, Ningbo, People’s Republic of China

    Yifu Gong

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  1. Wei Zhou
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  2. Chengtao Huang
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  3. Yifu Gong
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  4. Qili Feng
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  5. Feng Gao
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Correspondence to Feng Gao.

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W. Zhou and C. Huang contributed equally to this work.

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Zhou, W., Huang, C., Gong, Y. et al. Molecular Cloning and Expression Analysis of an ANS Gene Encoding Anthocyanidin Synthase from Purple-Fleshed Sweet Potato [Ipomoea batatas (L.) Lam]. Plant Mol Biol Rep 28, 112–121 (2010). https://doi.org/10.1007/s11105-009-0133-0

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