biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 51:277-284, 2007 | DOI: 10.1007/s10535-007-0055-3

Improvement of protein quality in transgenic soybean plants

H. A. El-Shemy1,2,*, M. M. Khalafalla2,3, K. Fujita4, M. Ishimoto2
1 Biochemistry Department, Faculty of Agriculture, Cairo University, Giza, Egypt
2 National Agricultural Research Center for Western Region, Fukuyama, Hiroshima, Japan
3 Commission of Biotechnology and Genetic Engineering, National Center for Research, Khartoum, Sudan
4 Graduate School of Biosphere Sciences, Hiroshima University, Higashi-Hiroshima, Japan

Glycinin is one of the abundant storage proteins in soybean seeds. A modified Gy1 (A1aB1b) proglycinin gene with a synthetic DNA encoding four continuous methionines (V3-1) was connected between the hpt gene and the modified green fluorescent protein sGFP(S65T) gene, and a resultant plasmid was introduced into soybean by particle bombardment in order to improve nutritional value of its seeds. After the selection with hygromycin, the efficiency of gene introduction was evaluated. More than 60 % of the regenerated plants tolerant to hygromycin yielded the hpt and V3-1 fragment by polymerase chain reaction (PCR) analysis, and the expression of sGFP was detected in about 50 % of putative transgenic soybeans. Southern hybridization confirmed the presence of transgenes in T0 plants and the transgenic soybeans hybridized with the hpt and V3-1 genes were analyzed showed different banding patterns. Most of the transgenic plants were growing, flowering normally and produced seeds. Analysis of seed obtained from transgenic soybean plants expressing hpt and V3-1 genes showed higher accumulation of glycinin compared with non-transgenic plants. In addition, protein expression in transgenic soybean plants was observed by using 2D-electrophoresis.

Keywords: genetic improvement; Glycine max; glycinin; particle bombardment; PCR; seed storage proteins
Subjects: chimeric genes; germination; Glycine max; glycinin; green fluorescent protein (GFP); in vitro culture, embryogenesis, somatic embryogenesis; in vitro culture, regeneration, proliferation, differentiation; particle bombardment; plasmid; polymerase chain reaction (PCR); seed storage; Southern blot analysis; soybean; transgenic plants

Received: September 13, 2005; Accepted: April 3, 2006; Published: June 1, 2007  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
El-Shemy, H.A., Khalafalla, M.M., Fujita, K., & Ishimoto, M. (2007). Improvement of protein quality in transgenic soybean plants. Biologia plantarum, 51(2), 277-284. doi: 10.1007/s10535-007-0055-3
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. Adachi, M., Kanamori, J., Masuda, T., Yagasaki, K., Kitamura, K., Mikami, B., Utsumi, S.: Crystal structure of soybean 11S globulin: glycinin A3B4 homohexamer.-Proc. nat. Acad. Sci. USA 100: 7395-7400, 2003. Go to original source...
    2. Adessi, C., Miege, C., Albrieux, C., Rabilloud, T.: Two-dimensional electrophoresis of membrane proteins: a current challenge for immobilized pH gradients.-Electrophoresis 18: 127-135, 2001. Go to original source...
    3. Altenbach, S.B., Pearson, K.W., Leung, F.W. Sun, S.S.M.: Cloning and sequence analysis of a cDNA encoding a Brazil nut protein exceptionally rich in methionine.-Plant mol. Biol. l8: 239-250, 1987. Go to original source...
    4. Altenbach, S.B., Pearson, K.W., Meeker, G., Staraci, L.C., Sun, S.S.M.: Enhancement of the methionine content of seed proteins by the expression of a chimeric gene encoding a methionine-rich protein in transgenic plants.-Plant mol. Biol. 13: 513-522, 1989. Go to original source...
    5. Chiu, W.-L., Niwa, Y., Zeng, W., Hirano, T., Kobayashi, H., Sheen, J.: Engineered GFP as a vital reporter in plants.-Curr. Biol. 6: 325-330, 1996. Go to original source...
    6. Draper, J., Scott, R. (ed.): The Isolation of Plant Nucleic Acids.-Blackwell Scientific Publications, London 1988.
    7. El-Shemy, H.A., Khalafalla, M.M., Nishizawa, K., Utsumi, S., Ishimoto, M.: Isolation of soybean plants with stable transgene expression by visual selection based on green fluorescent protein.-Mol. Breed. 14: 227-238, 2004. Go to original source...
    8. Finer, J.J., Nagasawa, A.: Development of an embryogenic suspension culture of soybean [Glycine max (L.) Merrill.].-Plant Cell Tissue Organ Cult. 15: 125-136, 1988. Go to original source...
    9. Finer, J.J., McMullen, M.D.: Transformation of soybean via particle bombardment of embryogenic suspention culture tissue.-In vitro cell. dev. Biol. Plant 27: 175-182, 1991. Go to original source...
    10. Finnie, C., Melchior, S., Roepstorff, P., Svensson, B.: Proteome analysis of grain filling and seed maturation in barley.-Plant Physiol. 129: 1308-1319, 2002. Go to original source...
    11. Gamborg, O.L., Miller, R.A., Ojima, K.: Nutrient requirements of suspension cultures of soybean root cells.-Exp. Cell Res. 50: 151-158, 1968. Go to original source...
    12. Gorg, A., Obermaier, C., Boguth, G., Csordas, A., Diaz, J., Madjar, J.: Very alkaline immobilized pH gradients for two-dimensional electrophoresis of ribosomal and nuclear proteins.-Electrophoresis 18: 328-337, 1997. Go to original source...
    13. Hadi, M.Z., McMullen, M.D., Finer, J.J.: Transformation of 12 different plasmids into soybean via particle bombardment.-Plant Cell Rep. 15: 500-505, 1996. Go to original source...
    14. Hajduch, M., Tanaka, H., Morinaka, Y., Otake, Y., Hakamura, H., Kayano, T., Kaga-Ban, Y.: Protein analysis of dwarfed transgenic rice plants overexpressing GA2-oxidase gene.-Biol. Plant. 49: 621-624, 2005. Go to original source...
    15. Hajika, M., Takahashi, M., Sakai, S., Igita, M.: A new genotype of 7S globulin (beta;-conglycinin) detected in wild soybean (Glycine soja Sieb. et Zucc.).-Breed. Sci. 46: 385-386, 1996. Go to original source...
    16. Harada, K., Toyokawa, Y., Kitamura, K.: Genetic analysis of the most acidic 11S globulin subunit and related characters in soybean seeds.-Jap. J. Breed. 33: 23-30, 1983. Go to original source...
    17. Herman, E.M., Helm, R.M., Jung, R., Kinney, A.J.: Genetic modification removes an immunodominant allergen from soybean.-Plant Physiol. 132: 36-43, 2003. Go to original source...
    18. Hinchee, M.A.W., Conner-Ward, D.V., Newell, C.A., McDonnell, R.E., Sato, S.J., Gasser, C.S., Fischhoff, D.A., Re, D.B., Fraley, R.T., Horsch, R.B.: Production of transgenic soybean plants using Agrobacterium-mediated DNA transfer.-Bio/Technology 6: 915-922, 1988. Go to original source...
    19. Hu, B., Esen, A.: Heterogeneity of soybean proteins: two-dimensional electrophoretic maps of three solubility fractions.-J. Agr. Food Chem. 30: 21-25, 1982. Go to original source...
    20. Ikeda, T.M., Ohnishi, N., Nagamine, T., Oda, S., Hisatomi, T., Yano, H.: Identification of new puroindoline genotypes and their protein products among wheat cultivars.-J. cereal Sci. 4: 1-6, 2005. Go to original source...
    21. Kim, C.S., Kamiya, S., Sato, T., Utsumi, S., Kito, M.: Improvement of nutritional value and functional properties of soybean glycinin by protein engineering.-Protein Eng. 3: 725-731, 1990. Go to original source...
    22. Kitamura, K., Davies, C.S., Nielsen, N.C.: Inheritance of alleles for Cgy1 and Gy4 storage protein genes in soybean.-Theor. appl. Genet. 68: 253-257, 1984. Go to original source...
    23. Kitamura, Y., Arahira, M., Itoh, Y., Fukazawa, C.: The complete nucleotide sequence of soybean glycinin A2B1a gene spanning to another glycinin gene A1aB1b.-Nucl. Acids Res. 18: 4245, 1990. Go to original source...
    24. Koshiyama, I.: Chemical and physical properties of a 7S protein in soybean globulins.-Cereal Chem. 45: 394-404, 1968.
    25. Laemmli, U.K.: Cleavage of structural proteins during the assembly of the head bacteriophage T4.-Nature 277: 680-685, 1970. Go to original source...
    26. Li, H.Y., Zhu, Y.M., Chen, Q., Conner, R.L., Ding, X.D., Li, J., Zhang, B.B.: Production of transgenic soybean plants with two antifungal protein genes via Agrobacterium and particle bombardment.-Biol. Plant. 48: 367-374, 2004. Go to original source...
    27. Maruyama, N., Mohamed Salleh, M.R., Takahashi, K., Yagasaki, K., Goto, H., Hontani, N., Nakagawa, S., Utsumi, S.: Structure-physicochemical function relationships of soybean beta-conglycinin heterotrimers.-J. Agr. Food Chem. 50: 4323-4326, 2002. Go to original source...
    28. Mooney, B.P., Thelen, J.J.: High-throughput peptide mass fingerprinting of soybean seed proteins: automated workflow and utility of UniGene expressed sequence tag databases for protein identification.-Phytochemistry 65: 1733-1744, 2004. Go to original source...
    29. Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassays with tobacco tissue culture.-Physiol. Plant. 15: 473-497, 1962. Go to original source...
    30. Natarajan, S., Xu, C., Caperna, T J., Garrett, W M.: Comparison of protein solubilization methods suitable for proteomic analysis of soybean seed proteins.-Anal. Biochem. 342: 214-220, 2005. Go to original source...
    31. Neuhoff, V., Arold, N., Taube, D., Ehrhardt, W.: Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Blue G-250 and R-250.-Electrophoresis 9: 255-262, 1988. Go to original source...
    32. Nielsen, N.C.: Structure and complexity of the 11S polypeptides in soybean.-J. amer. Oil Chem. Soc. 62: 1680-1686, 1985. Go to original source...
    33. Nordlee, J.A., Taylor, S.L., Townsend, J.A., Thomas, L.A., Bush, R.K.: Identification of a brazil-nut allergen in transgenic soybean.-New Eng. J. Med. 334: 688-692, 1996. Go to original source...
    34. Odanaka, H., Kaizuma, N.: Mutants on soybean storage proteins induced with gama-ray irradiation.-Jap. J. Breed. 39: 430-431, 1989.
    35. Ogawa, T., Tayama, E., Kitamura, K., Kaizuma, N.: Genetic improvement of seed storage proteins using three varient alleles of 7S globulin subunits in soybean.-Jap. J. Breed. 39: 137-147, 1989. Go to original source...
    36. Russel, D.R., Wallace, K.M., Bathe, J.H., Martinell, B.J., McCabe, D.E.: Stable transformation of Phaseolus vulgaris via electric-discharge mediated particle acceleration.-Plant Cell Rep. 12: 165-169, 1993. Go to original source...
    37. Saalbach, I., Pickardt, T., Machemehl, F., Saalbach, G., Schieder, O., Müntz, K.: A chimeric gene encoding the methionine-rich 2S albumin of the Brazil nut (Bertholletia excelsa H.B.K.) is stably expressed and inherited in transgenic grain legumes.-Mol. gen. Genet. 242: 226-236, 1994. Go to original source...
    38. Saalbach, I., Waddell, D., Pickardt, T., Schieder, O., Müntz, K.: Stable expression of the sulfur-rich 2S albumin gene in transgenic Vicia narbonensis increases the methionine content of seed.-J. Plant Physiol. 145: 674-681, 1995. Go to original source...
    39. Samoylov, V.M., Tucker, D.M., Parrott, W.A.: A liquid medium-based protocol for rapid regeneration from embryogenic soybean cultures.-Plant Cell Rep. 18: 49-54, 1998. Go to original source...
    40. Sato, S., Newell, C., Kolacz, K., Tredo, L., Finer, J., Hinchee, M.: Stable transformation via particle bombardment in two different soybean regeneration systems.-Plant Cell Rep. 12: 408-413, 1993. Go to original source...
    41. Schroeder, H.E., Schotz, A.H., Wardley-Richardson, T., Spencer, D., Higgins, T.J.V.: Transformation and regeneration of two cultivars of pea (Pisum sativum L.).-Plant Physiol. 101: 751-757, 1993. Go to original source...
    42. Staswick, P.E., Hermodson, M.A., Nielsen, N.C.: Identification of the cystines which link the acidic and basic components of the glycinin subunits.-J. biol. Chem. 259: 13431-13435, 1984. Go to original source...
    43. Takahashi, K., Banaba, H., Kikuchi, A., Ito, M., Nakamura, S.: An iduced mutant line lacking the α-subunit of β-conglycinin in soybean (Glycine max (L.) Merrill).-Breed. Sci. 46: 251-255, 1994. Go to original source...
    44. Yagasaki, K., Kaizuma, N., KItamura, K.: Inheritance of glynicin subunits and characterization of glycinin molecules lacking the subunits in soybean (Glycine max (L.) Merr.).-Breed. Sci. 46: 11-15, 1996. Go to original source...
    45. Yamada, T., Teraishi, M., Hattori, K., Ishimoto, M.: Transformation of azuki bean by Agrobacterium tumefaciens.-Plant Cell Tissue Organ Cult. 64: 47-54, 2001. Go to original source...

    Return to the content