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Assessment of conditions affecting Agrobacterium-mediated soybean transformation using the cotyledonary node explant

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Abstract

Conditions affecting Agrobacterium-mediated transformation of soybean [Glycine max (L.) Merr.], including seed vigor of explant source, selection system, and cocultivation conditions, were investigated. A negative correlation between seed sterilization duration and seed vigor, and a positive correlation between seed vigor and regenerability of explants were observed in the study, suggesting that use of high vigor seed and minimum seed sterilization duration can further improve transformation efficiency. Selection schemes using glufosinate or bialaphos as selective agents in vitro were assessed. Glufosinate selection enhanced soybean transformation as compared to bialaphos. The use of 6 mg L-1 glufosinate during shoot induction and shoot elongation stages yielded higher final transformation efficiency ranging from 2.0% to 6.3% while bialaphos at 4 to 6 mg L-1 gave 0% to 2.1% efficiency. Including cysteine and DTT during cocultivation increased the transformation efficiency from 0.2–0.9% to 0.6–2.9%. This treatment also improved T-DNA transfer as indicated by enhanced transient GUS expression. Shoot regeneration and Agrobacterium infection were attained in twelve soybean cultivars belonging to maturity groups I-VI. These cultivars maybe amenable to genetic transformation and may provide a valuable tool in soybean improvement programs.

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Authors and Affiliations

  1. Plant Transformation Facility, Department of Agronomy, Iowa State University, Ames, IA, 50011–1010, USA

    Margie M. Paz, Huixia Shou, Zibiao Guo, Zhanyuan Zhang, Anjan K. Banerjee & Kan Wang

  2. College of Life Science, Zhejiang University, Hangzhou, 310029, China

    Huixia Shou

  3. Plant Biology Division, The Samuel Roberts Noble Foundation, Inc., 2510 Sam Noble Parkway, Ardmore, OK, 73401, USA

    Zibiao Guo

  4. Plant Transformation Core Facility, University of Missouri-Columbia, MO, 65211, USA

    Zhanyuan Zhang

  5. Department of Horticulture, Iowa State University, Ames, IA, 50011-1100, USA

    Anjan K. Banerjee

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  1. Margie M. Paz
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Paz, M.M., Shou, H., Guo, Z. et al. Assessment of conditions affecting Agrobacterium-mediated soybean transformation using the cotyledonary node explant. Euphytica 136, 167–179 (2004). https://doi.org/10.1023/B:EUPH.0000030669.75809.dc

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