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Fructan:fructan 1-fructosyltransferase, a key enzyme for biosynthesis of graminan oligomers in hardened wheat

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Abstract

Fructans play important roles not only as a carbon source for survival under persistent snow cover but also as agents that protect against various stresses in overwintering plants. Complex fructans having both ß-(2,1)- and ß-(2,6)-linked fructosyl units accumulate in wheat (Triticum aestivum L.) during cold hardening. We detected fructan: fructan 1-fructosyltransferase (1-FFT; EC 2.4.1.100) activity for catalyzing the formation and extension of ß-(2,1)-linked fructans in hardened wheat tissues, cloned cDNAs (wft3 and wft4) of 1-FFT, and analyzed the enzymatic properties of a wft3 recombinant protein (Wft3m) produced by yeast. Wft3m transferred ß-(2,1)-linked fructosyl units to phlein, an extension of sucrose through ß-(2,6)-linked fructosyl units, as well as to inulin, an extension of sucrose through ß-(2,1)-linked fructosyl units, but could not efficiently synthesize long inulin oligomers. Incubation of a mixture of Wft3m and another recombinant protein of wheat, sucrose:fructan 6-fructosyltransferase (6-SFT), with sucrose and 1-kestotriose produced fructans similar to those that accumulated in hardened wheat tissues. The results demonstrate that 1-FFT produces branches of ß-(2,1)-linked fructosyl units to phlein and graminan oligomers synthesized by 6-SFT and contributes to accumulation of fructans containing ß-(2,1)- and ß-(2,6)-linked fructosyl units. In combination with sucrose:sucrose 1-fructosyltransferase (1-SST; EC 2.4.1.99) and 6-SFT, 1-FFT is necessary for fructan synthesis in hardened wheat.

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Abbreviations

1-SST:

Sucrose:sucrose 1fructosyltransferase

1-FFT:

Fructan:fructan 1-fructosyltransferase

6-SFT:

Sucrose:fructan 6-fructosyltransferase

6G-FFT:

Fructan:fructan 6G-fructosyltransferase

FEH:

Fructan exohydrolase

HPAEC:

High-performance anion exchange chromatography

PAD:

Pulsed amperometric detector

DP:

Degree of polymerization

Wft1:

Wheat fructosyltransferase 1

Wft3:

Wheat fructosyltransferase 3

Wft4:

Wheat fructosyltransferase 4

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Acknowledgements

We thank Drs. Himi and Noda, Okayama University, Japan for providing the sequence of wheat actin cDNA, Dr. Masaru Iizuka, Osaka City University, Japan for providing 6-kestose, and Dr Wim Van den Ende, University of Leuven, Belgium for providing inulin extracted from chicory.

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Authors and Affiliations

  1. Laboratory of Fungal Disease, Department of Plant Pathology, National Agricultural Research Center, Kannondai, Tsukuba, Ibaraki, 305-8666, Japan

    Akira Kawakami

  2. Laboratory of Winter Stress, Department of Low Temperature Science, National Agricultural Research Center for Hokkaido Region, Uitsujigaoka, Sapporo, Hokkaido, 062-8555, Japan

    Midori Yoshida

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  1. Akira Kawakami
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  2. Midori Yoshida
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Correspondence to Akira Kawakami.

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Kawakami, A., Yoshida, M. Fructan:fructan 1-fructosyltransferase, a key enzyme for biosynthesis of graminan oligomers in hardened wheat. Planta 223, 90–104 (2005). https://doi.org/10.1007/s00425-005-0054-6

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  • DOI: https://doi.org/10.1007/s00425-005-0054-6

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