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Transgenic approach to improve wheat (Triticum aestivum L.) nutritional quality

  • Genetic Transformation and Hybridization
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Abstract

An amaranth (Amaranthus hypochondriacus) albumin gene, encoding the 35-kDa AmA1 protein of the seed, with a high content of essential amino acids, was used in the biolistic transformation of bread wheat (Triticum aestivum L.) variety Cadenza. The transformation cassette carried the ama1 gene under the control of a powerful wheat endosperm-specific promoter (1Bx17 HMW-GS). Southern-blot analysis of T1 lines confirmed the integration of the foreign gene, while RT-PCR and Western-blot analyses of the samples confirmed the transcription and translation of the transgene. The effects of the extra albumin protein on the properties of flour, produced from bulked T2 seeds, were calculated using total protein and essential amino acid content analysis, polymeric/monomeric protein and HMW/LMW glutenin subunit ratio measurements. The results indicated that not only can essential amino acid content be increased, but some parameters associated with functional quality may also be improved because of the expression of the AmA1 protein.

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Abbreviations

RT-PCR:

Reverse transcription polymerase chain reaction

HMW-GS:

High-molecular-weight glutenin subunit

LMW-GS:

Low-molecular-weight glutenin subunit

DPA:

Day post-anthesis

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

BCIP:

5-Bromo-4-chloro-3-indolyl phosphate

NBT:

Nitro blue tetrazolium chloride

DTT:

1, 4-Dithiotreitol

RP-HPLC:

Reverse phase high-performance liquid chromatography

SE-HPLC:

Size exclusion high-performance liquid chromatography

UPP:

Unextractable polymeric protein

HRP:

Horseradish peroxidase

PBST:

Phosphate buffered saline

OD:

Optical density

GUS:

β-Glucuronidase

CTAB:

Cetyl trimethylammonium bromide

PVDF:

Polyvinylidene fluoride

LSD:

Least significant difference

GBSS:

Granule-bound starch synthase

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Acknowledgments

This work was partly supported by the Hungarian National Scientific Research Fund (OTKA T 034791 and K 67844) and by the Bilateral Intergovernmental Science and Technology Cooperation (KOR 13/99). The authors are grateful to Dr. Ferenc Békés for help with the statistical analyses and for his critical reading of the manuscript and valuable discussions. The contributions of Dr. Tae-Jin Kang and Dr. Péter Szűcs are also appreciated to this work. Thanks are due to Dr. Maria Oszvald for help with the measurement of flour protein content and to Dr. Sándor Tömösközi and Gábor Balázs (BUTE, Budapest) for the HPLC study. The authors wish to acknowledge the excellent technical assistance of Mrs Magdolna P Holtai.

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Authors and Affiliations

  1. Agricultural Research Institute of the Hungarian Academy of Sciences, 2462, Martonvásár, Hungary

    Cecília Tamás, Boglárka N. Kisgyörgy, Mariann Rakszegi, László Láng & Zoltán Bedő

  2. Plant Science Department, Rothamsted Research, Harpenden, Hertfordshire, AL5 2JQ, UK

    Mark D. Wilkinson

  3. Chonbuk National University, Jonjou, South-Korea

    Moon-Sik Yang

  4. Department of Plant Physiology, Eötvös Loránd University, 1117, Budapest, Hungary

    László Tamás

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  1. Cecília Tamás
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  2. Boglárka N. Kisgyörgy
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  3. Mariann Rakszegi
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  4. Mark D. Wilkinson
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  5. Moon-Sik Yang
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  6. László Láng
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  7. László Tamás
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  8. Zoltán Bedő
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Correspondence to László Tamás.

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Communicated by H. Jones.

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Tamás, C., Kisgyörgy, B.N., Rakszegi, M. et al. Transgenic approach to improve wheat (Triticum aestivum L.) nutritional quality. Plant Cell Rep 28, 1085–1094 (2009). https://doi.org/10.1007/s00299-009-0716-0

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