Plant Cell Reports

, Volume 25, Issue 3, pp 206–213 | Cite as

Improved cotyledonary node method using an alternative explant derived from mature seed for efficient Agrobacterium-mediated soybean transformation

  • Margie M. Paz
  • Juan Carlos Martinez
  • Andrea B. Kalvig
  • Tina M. Fonger
  • Kan Wang
Genetic Transformation and Hybridization

Abstract

The utility of transformation for soybean improvement requires an efficient system for production of stable transgenic lines. We describe here an improved cotyledonary node method using an alternative explant for Agrobacterium tumefaciens-mediated soybean transformation. We use the term “half-seed” to refer to this alternative cotyledonary explant that is derived from mature seed of soybean following an overnight imbibition and to distinguish it from cotyledonary node derived from 5–7-day-old seedlings. Transformation efficiencies using half-seed explants ranged between 1.4 and 8.7% with an overall efficiency of 3.8% based on the number of transformed events that have been confirmed in the T1 generation by phenotypic assay using the herbicide Liberty® (active ingredient glufosinate) and by Southern analysis. This efficiency is 1.5-fold higher than the cotyledonary node method used in our laboratory. Significantly, the half-seed system is simple and does not require deliberate wounding of explants, which is a critical and technically demanding step in the cotyledonary node method.

Keywords

Agrobacterium tumefaciens Cotyledonary node Glufosinate Half-seed Soybean Transformation 

Notes

Acknowledgements

The authors thank Joanna Harbaugh, Amanda Ehrler and Francois Torney for their technical assistance; Mike Webber, Bayer CropScience, USA for graciously giving us Liberty® herbicide; and Bronwyn Frame and John Pesek for critical review of the manuscript and valuable suggestions. This work was partially supported by the Iowa Soybean Promotion Board and the North Central Soybean Research Program.

References

  1. An G, Evert PR, Mitra A, Ha SB (1988) Binary vectors. In: Gelvin SB, Schilperoort RA (eds.) Plant molecular biology manual A3. Kluwer Academic Publishers, Dordrecht, pp 1–19Google Scholar
  2. Buising CM, Shoemaker RC, Benbow RM (1994) Early events of multiple bud formation and shoot development in soybean embryonic axes treated with the cytokinin, 6-benzylaminopurine. Am J Bot 81:1435–1448CrossRefGoogle Scholar
  3. Carrington JC, Freed DD (1990) Cap-independent enhancement of translation by a plant potyvirus 5′ nontranslated region. J Virol 64:1590–1597PubMedGoogle Scholar
  4. Clemente T, LaVallee BJ, Howe AR, Ward DC, Rozman RJ, Hunter PE, Broyles DL, Kasten DS, Hinchee MA (2000) Progeny analysis of glyphosate selected transgenic soybeans derived from Agrobacterium-mediated transformation. Crop Sci 40:797–803CrossRefGoogle Scholar
  5. Dan Y, Reichert NA (1998) Organogenic regeneration of soybean from hypocotyl explants. In Vitro Cell Dev Biol-Plant 34:14–21Google Scholar
  6. Di R, Purcell V, Collins GB, Ghabril SA (1996) Production of transgenic soybean lines expressing the bean pod mottle virus coat protein precursor gene. Plant Cell Rep 15:746–750CrossRefGoogle Scholar
  7. Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15Google Scholar
  8. Droste A, Pasquali G, Bodanese-Zanettini MH (2000) Integrated bombardment and Agrobacterium transformation system: an alternative method for soybean transformation. Plant Mol Biol Rep 18:51–59CrossRefGoogle Scholar
  9. Droste A, Pasquali G, Bodanese-Zanettini MH (2002) Transgenic fertile plants of soybean [Glycine max (L.) Merrill] obtained from bombarded embryogenic tissue. Euphytica 127:367–376CrossRefGoogle Scholar
  10. Finer KR, Finer JJ (2000) Use of Agrobacterium expressing green fluorescent protein to evaluate colonization of sonication-assisted Agrobacterium-mediated transformation-treated soybean cotyledons. Lett Appl Microbiol 30:406–410CrossRefPubMedGoogle Scholar
  11. Frame BR, Shou H, Chikwamba RK, Zhang Z, Xiang C, Fonger TM, Pegg SEK, Li B, Nettleton DS, Pei D, Wang K (2002) Agrobacterium tumefaciens-mediated transformation of maize embryos using a standard binary vector system. Plant Physiol 129:13–22CrossRefPubMedGoogle Scholar
  12. Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158CrossRefPubMedGoogle Scholar
  13. Hadi MZ, McMullen MD, Finer JJ (1996) Transformation of 12 different plasmids into soybean via particle bombardment. Plant Cell Rep 15:500–505CrossRefGoogle Scholar
  14. Hajdukiewicz P, Svab Z, Maliga P (1994) The small versatile pZP family of Agrobacterium binary vectors for plant transformation. Plant Mol Biol 25:989–994PubMedCrossRefGoogle Scholar
  15. Hinchee MAW, Connor-Ward DV, Newell CA, McDonnell RE, Sato SJ, Gasser CS, Fischhoff DA, Re DB, Fraley RT, Horsch RB (1988) Production of transgenic soybean plants using Agrobacterium-mediated DNA transfer. Biotechnology 6:915–922CrossRefGoogle Scholar
  16. Hood EE, Helmer GL, Fraley RT, Chilton MD (1986) The hypervirulence of Agrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside of T-DNA. J Bacteriol 168:1291–1301PubMedGoogle Scholar
  17. Ko T, Lee S, Krasnyanski S, Korban SS (2003) Two critical factors are required for efficient transformation of multiple soybean cultivars: Agrobacterium strain and orientation of immature cotyledonary explant. Theor Appl Genet 107:439–447CrossRefPubMedGoogle Scholar
  18. Koscianska E, Wypijewski K (2001) Electroporated intact BY-2 tobacco culture cells as a model of transient expression study. Acta Biochim Pol 48:657–661PubMedGoogle Scholar
  19. Liu H, Yang C, Wei Z (2004) Efficient Agrobacterium tumefaciens-mediated transformation of soybeans using an embryonic tip regeneration system. Planta 219:1042–1049CrossRefPubMedGoogle Scholar
  20. Mason HS, DeWald D, Mullet JE (1993) Identification of a methyl jasmonate-responsive domain in the soybean vspB promoter. Plant Cell 5:241–251CrossRefPubMedGoogle Scholar
  21. Meurer CA, Dinkins RD, Collins GB (1998) Factors affecting soybean cotyledonary node transformation. Plant Cell Rep 18:180–186CrossRefGoogle Scholar
  22. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–479CrossRefGoogle Scholar
  23. Odell JT, Nagy F, Chua NH (1985) Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. Nature 6:810–812CrossRefGoogle Scholar
  24. Olhoft PM, Flagel LE, Donovan CM, Somers DA (2003) Efficient soybean transformation using hygromycin B selection in the cotyledonary-node method. Planta 216:723–735PubMedGoogle Scholar
  25. Olhoft PM, Lin K, Galbraith J, Nielsen NC, Somers DA (2001) The role of thiol compounds in increasing Agrobacterium-mediated transformation of soybean cotyledonary-node cells. Plant Cell Rep 20:731–737CrossRefGoogle Scholar
  26. Parrott WA, Hoffman LM, Hildebrand DF, Williams EG, Collins GB (1989) Recovery of primary transformants of soybean. Plant Cell Rep 7:615–617Google Scholar
  27. Paz MM, Shou H, Guo Z, Zhang Z, Banerjee AK, Wang K (2004) Assessment of conditions affecting Agrobacterium-mediated soybean transformation using the cotyledonary node explant. Euphytica 136:167–179CrossRefGoogle Scholar
  28. Saka H, Voqui-Dinh TH, Cheng T (1980) Stimulation of multiple shoot formation on soybean stem nodes in culture. Plant Sci Lett 19:193–201CrossRefGoogle Scholar
  29. Santarem ER, Finer JJ (1999) Transformation of soybean [Glycine max (L.) Merrill] using proliferative embryogenic tissue maintained on semi-solid medium. In Vitro Cell Dev Biol Plant 35:451–455CrossRefGoogle Scholar
  30. Santarem ER, Trick HN, Essig JS, Finer JJ (1998) Sonication-assisted Agrobacterium-mediated transformation of soybean immature cotyledons: optimization of transient expression. Plant Cell Rep 17:752–759CrossRefGoogle Scholar
  31. Somers DA, Samac DA, Olhoft PM (2003) Recent advances in legume transformation. Plant Physiol 131:892–899CrossRefPubMedGoogle Scholar
  32. Townsend JA, Thomas LA (1993) An improved method of Agrobacterium-mediated transformation of cultured soybean cells. US Patent WO 94/02620Google Scholar
  33. Vancanneyt G, Schmidt R, O'Connor-Sanchez A, Willmitzer L, Rocha-Sosa M (1990) Construction of an intron-containing marker gene: splicing of the intron in transgenic plants and its use in monitoring early events in Agrobacterium-mediated plant transformation. Mol Gen Genet 220:245–250CrossRefPubMedGoogle Scholar
  34. White J, Chang SP, Bibb MJ, Bibb MJ (1990) A cassette containing the bar gene of Streptomyces hygroscopicus: a selectable marker for plant transformation. Nucl Acids Res 18:1062PubMedCrossRefGoogle Scholar
  35. Wright MS, Koehler SM, Hinchee MA, Carnes MG (1986) Plant regeneration by organogenesis in Glycine max. Plant Cell Rep 5:150–154CrossRefGoogle Scholar
  36. Wroblewski T, Tomczak A, Michelmore R (2005) Optimization of Agrobacterium-mediated transient assays of gene expression in lettuce, tomato and Arabidopsis. Plant Biotechnol J 3:259–273CrossRefPubMedGoogle Scholar
  37. Wu FS, Feng TY (1999) Delivery of plasmid DNA into intact plant cells by electroporation of plasmolyzed cells. Plant Cell Rep 18:381–386CrossRefGoogle Scholar
  38. Yan B, Reddy MSS, Collins GB, Dinkins RD (2000) Agrobacterium tumefaciens– mediated transformation of soybean [Glycine max (L.) Merrill] using immature zygotic cotyledon explants. Plant Cell Rep 19:1090–1097CrossRefGoogle Scholar
  39. Zeng P, Vadnais DA, Zhang Z, Polacco JC (2004) Refined glufosinate selection in Agrobacterium-mediated transformation of soybean [Glycine max (L.) Merrill]. Plant Cell Rep 22:478–482CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Margie M. Paz
    • 1
  • Juan Carlos Martinez
    • 1
  • Andrea B. Kalvig
    • 1
  • Tina M. Fonger
    • 1
  • Kan Wang
    • 1
  1. 1.Department of AgronomyIowa State UniversityAmesUSA

Personalised recommendations