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Accumulation of multiple-repeat starch-binding domains (SBD2–SBD5) does not reduce amylose content of potato starch granules

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Abstract

This study investigates whether it is possible to produce an amylose-free potato starch by displacing the amylose enzyme, granule-bound starch synthase I (GBSSI), from the starch granule by engineered, high-affinity, multiple-repeat family 20 starch-binding domains (SBD2, SBD3, SBD4, and SBD5). The constructs were introduced in the amylose-containing potato cultivar (cv. Kardal), and the starches of the resulting transformants were compared with those of SBD2-expressing amylose-free (amf) potato clones. It is shown that a correctly sized protein accumulated in the starch granules of the various transformants. The amount of SBD accumulated in starch increased progressively from SBD to SBD3; however, it seemed as if less SBD4 and SBD5 was accumulated. A reduction in amylose content was not achieved in any of the transformants. However, it is shown that SBDn expression can affect physical processes underlying granule assembly, in both genetic potato backgrounds, without altering the primary structure of the constituent starch polymers and the granule melting temperature. Granule size distribution of the starches obtained from transgenic Kardal plants were similar to those from untransformed controls, irrespective of the amount of SBDn accumulated. In the amf background, granule size is severely affected. In both the Kardal and amf background, apparently normal oval-shaped starch granules were composed of multiple smaller ones, as evidenced from the many “Maltese crosses” within these granules. The results are discussed in terms of different binding modes of SBD.

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Abbreviations

SBD:

Starch-binding domain

SBD2:

Double starch-binding domain

GBSSI:

Granule-bound starch synthase I

amf :

Amylose-free

LM:

Light microscopy

SEM:

Scanning electron microscopy

DSC:

Differential scanning calorimetry

KD-UT:

Kardal untransformed

amf-UT:

Amylose-free mutant untransformed

KDS:

Kardal transformant containing SBD gene

KDSS:

Kardal transformant containing double SBD gene

KDS3:

Kardal transformant containing triple SBD gene

KDS4:

Kardal transformant containing quadruple SBD gene

KDS5:

Kardal transformant containing pentuple SBD gene

amfS:

amf mutant transformed with SBD gene

amfSS:

amf mutant transformed with double SBD gene

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Authors and Affiliations

  1. Graduate School Experimental Plant Sciences, Laboratory of Plant Breeding, Wageningen University, P.O. Box 386, 6700 AJ, Wageningen, The Netherlands

    Farhad Nazarian Firouzabadi, Jean-Paul Vincken, Qin Ji, Luc C. J. M. Suurs & Richard G. F. Visser

  2. Agronomy and plant breeding group, Faculty of Agriculture, University of Lorestan, P.O. Box 465, Khorramabad, Iran

    Farhad Nazarian Firouzabadi

  3. Laboratory of Food Chemistry, Wageningen University, P.O. Box 8129, 6700 EV, Wageningen, The Netherlands

    Jean-Paul Vincken

  4. Department of Biology, HuaiYin Teachers College, 223300, Huaian, China

    Qin Ji

  5. Laboratoire de Physico-chimie des Macromolécules, Institut National de la Recherche Agronomique, BP 71627, 44316, Nantes Cedex 3, France

    Alain Buléon

Authors
  1. Farhad Nazarian Firouzabadi
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  2. Jean-Paul Vincken
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  3. Qin Ji
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  4. Luc C. J. M. Suurs
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  5. Alain Buléon
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  6. Richard G. F. Visser
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Correspondence to Richard G. F. Visser.

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Firouzabadi, F.N., Vincken, JP., Ji, Q. et al. Accumulation of multiple-repeat starch-binding domains (SBD2–SBD5) does not reduce amylose content of potato starch granules. Planta 225, 919–933 (2007). https://doi.org/10.1007/s00425-006-0411-0

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  • DOI: https://doi.org/10.1007/s00425-006-0411-0

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