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The effect of co-cultivation and selection parameters on Agrobacterium-mediated transformation of Chinese soybean varieties

Plant Cell Reports volume 27, pages 489–498 (2008)Cite this article

Abstract

In the present study, an efficient Agrobacterium-mediated gene transformation system was developed for soybean [Glycine max (L.) Merrill] based on the examinations of several factors affecting plant transformation efficiency. Increased transformation efficiencies were obtained when the soybean cotyledonary node were inoculated with the Agrobacterium inoculum added with 0.02% (v/v) surfactant (Silwet L-77). The applications of Silwet L-77 (0.02%) during infection and l-cysteine (600 mg l−1) during co-cultivation resulted in more significantly improved transformation efficiency than each of the two factors alone. The optimized temperature for infected explant co-cultivation was 22°C. Regenerated transgenic shoots were selected and produced more efficiently with the modified selection scheme (initiation on shoot induction medium without hygromycin for 7 days, with 3 mg l−1 hygromycin for 10 days, 5 mg l−1 hygromycin for another 10 days, and elongation on shoot elongation medium with 8 mg l−1 hygromycin). Using the optimized system, we obtained 145 morphologically normal and fertile independent transgenic plants in five important Chinese soybean varieties. The transformation efficacies ranged from 3.8 to 11.7%. Stable integration, expression and inheritance of the transgenes were confirmed by molecular and genetic analysis. T1 plants were analyzed and transmission of transgenes to the T1 generation in a Mendelian fashion was verified. This optimized transformation system should be employed for efficient Agrobacterium-mediated soybean gene transformation.

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Abbreviations

BAP :

N6-Benzylaminopurine

GUS :

β-Glucuronidase

IAA :

Indole-3-acetic acid

IBA :

Indole-3-butyric acid

GA 3 :

Gibberellic acid

MES :

2-[N-Morpholino]ethanesulfonic acid

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Affiliations

  1. Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, 200032, Shanghai, China

    Sheng-Jun Liu, Zhi-Ming Wei & Jian-Qiu Huang

  2. Graduate School of the Chinese Academy of Sciences, 300 Fenglin Road, 200032, Shanghai, China

    Sheng-Jun Liu

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  1. Sheng-Jun Liu
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  2. Zhi-Ming Wei
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  3. Jian-Qiu Huang
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Correspondence to Jian-Qiu Huang.

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Communicated by J. Register.

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Liu, SJ., Wei, ZM. & Huang, JQ. The effect of co-cultivation and selection parameters on Agrobacterium-mediated transformation of Chinese soybean varieties. Plant Cell Rep 27, 489–498 (2008). https://doi.org/10.1007/s00299-007-0475-8

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  • DOI: https://doi.org/10.1007/s00299-007-0475-8

Keywords

  • Agrobacterium tumefaciens
  • Glycine max
  • cotyledonary node
  • Transformation