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Molecular Breeding

, Volume 25, Issue 2, pp 313–326 | Cite as

Evaluation of amino acid content and nutritional quality of transgenic soybean seeds with high-level tryptophan accumulation

  • Masao IshimotoEmail author
  • Shaikh M. Rahman
  • Moemen S. Hanafy
  • Mutasim M. Khalafalla
  • Hany A. El-Shemy
  • Yumi Nakamoto
  • Yoichi Kita
  • Kojiro Takanashi
  • Fumio Matsuda
  • Yoshihiro Murano
  • Tomoko Funabashi
  • Hisashi Miyagawa
  • Kyo Wakasa
Article

Abstract

Anthranilate synthase (AS) is a key regulatory enzyme in tryptophan (Trp) biosynthesis and is subject to feedback inhibition by Trp. The gene encoding a mutated feedback-resistant α subunit of rice AS (OASA1D) under the control of either a soybean glycinin gene promoter or the 35S promoter of cauliflower mosaic virus for seed-specific or constitutive expression, respectively, was introduced into soybean [Glycine max (L.) Merrill] by particle bombardment. A total of seven different transgenic lines that showed markedly increased accumulation of free Trp in their seeds were developed. The overproduction of free Trp was stably inherited in subsequent generations without any apparent detrimental effect on plant growth or reproduction. The total Trp content of transgenic seeds was also about twice that of nontransgenic seeds, whereas the amount of protein-bound Trp was not substantially affected by OASA1D expression. In spite of the marked increase in free Trp content, metabolic profiling by high-performance liquid chromatography coupled with mass spectrometry revealed little change in the amounts of other aromatic compounds in the transgenic seeds. We developed a rapid and feasible system based on farmed rainbow trout to evaluate the nutritional quality of a limited quantity of transgenic soybean seeds. Supplementation of fish food with OASA1D transgenic soybean seeds or with nontransgenic seeds plus crystalline Trp increased the growth rate of the farmed fish. These results indicate transformation with OASA1D is a reliable approach to improve the nutritional quality of soybean (or of other grain legumes) for human and animal food.

Keywords

Amino acid content Anthranilate synthase Feeding trial Seed quality Transgenic plant Tryptophan content 

Notes

Acknowledgments

This study was supported by CREST of the Japan Science and Technology Agency. We thank Yumi Naganuma and Midori Yokoyama for technical assistance. S.M.R., M.H., and M.M.K. were supported by postdoctoral fellowships from the Japan Society for the Promotion of Science. H.A.E.-S. was supported by a postdoctoral fellowship from the Japanese Science and Technology Corporation. K.T., F. M., and H.M. thank Phenomenome Discoveries Inc. for help with metabolic profiling analysis by Profiler M software.

Supplementary material

11032_2009_9334_MOESM1_ESM.doc (164 kb)
Supplementary material 1 (DOC 164 kb)
11032_2009_9334_MOESM2_ESM.pdf (92 kb)
Supplementary material 2 (PDF 93 kb)

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Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Masao Ishimoto
    • 1
    • 2
    • 3
    Email author
  • Shaikh M. Rahman
    • 1
    • 3
    • 8
  • Moemen S. Hanafy
    • 1
    • 2
    • 3
    • 9
  • Mutasim M. Khalafalla
    • 1
    • 3
    • 10
  • Hany A. El-Shemy
    • 1
    • 3
    • 11
  • Yumi Nakamoto
    • 1
    • 2
  • Yoichi Kita
    • 2
    • 4
  • Kojiro Takanashi
    • 5
  • Fumio Matsuda
    • 1
    • 12
  • Yoshihiro Murano
    • 6
  • Tomoko Funabashi
    • 6
  • Hisashi Miyagawa
    • 1
    • 5
  • Kyo Wakasa
    • 1
    • 7
  1. 1.Core Research for Evolutional Science and Technology (CREST)Japan Science and Technology AgencyKawaguchiJapan
  2. 2.National Agricultural Research Center for Hokkaido RegionHokkaidoJapan
  3. 3.National Agricultural Research Center for Western RegionHiroshimaJapan
  4. 4.Graduate School of AgricultureHokkaido UniversitySapporoJapan
  5. 5.Graduate School of AgricultureKyoto UniversityKyotoJapan
  6. 6.Central Research LaboratoryNisshin OilliO Group LtdYokosukaJapan
  7. 7.Tokyo University of AgricultureKanagawaJapan
  8. 8.Fukuoka Agricultural Research CenterFukuokaJapan
  9. 9.Plant Biotechnology DepartmentNational Research CenterCairoEgypt
  10. 10.Commission of Biotechnology and Genetic EngineeringKhartoumSudan
  11. 11.Faculty of AgricultureCairo UniversityGizaEgypt
  12. 12.RIKEN Plant Science CenterYokohamaJapan

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