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Improved soybean transformation for efficient and high throughput transgenic production

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Abstract

Although genetic transformation of soybean dates back to over two decades, the process remains inefficient. Here, we report the development of an organogenesis-based transformation method of soybean that resulted in an average transformation frequency of 18.7%. This improved method resorts to Agrobacterium-mediated transformation of the split-seed explant with an attached partial embryonic axis obtained from an imbibed seed. In addition to the split-seed explant, Agrobacterium strain and preparation were shown to be important for improved transformation. Transformation with Agrobacterium tumefaciens EHA105 generated higher transformation frequencies and number of low copy events compared to the strain EHA101. In this system, phosphinothricin acetyl transferase conferring tolerance to glufosinate was successfully employed for efficiently producing transgenic events. Around 48% of the T1 progeny was demonstrated to be heritable based on molecular analysis and screening with the herbicide Liberty®. This method was shown to be applicable to different genotypes and a few elite lines showed high transformation frequencies. This split-seed system with an attached partial embryonic axis serves not only as an efficient means for high throughput transgenic production for basic research studies but also for the commercial development of transgenic soybean products.

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Acknowledgements

The authors thank Dr Todd Jones for critically reviewing the manuscript and helpful suggestions.

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Authors and Affiliations

  1. Dow AgroSciences LLC, Trait Product Development, Indianapolis, IN, USA

    Dayakar Pareddy, Tatyana Minnicks, Lauren Clark, David Griffin, Pon Samuel, Kelley Smith & Rodrigo Sarria

  2. Dow AgroSciences LLC, Trait Product Development, West Lafayette, IN, USA

    Siva Chennareddy, Geny Anthony, Nagesh Sardesai, Tejinder Mall, Olga Karpova, Brandon Bishop & Nolan Shumway

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  1. Dayakar Pareddy
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  2. Siva Chennareddy
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  3. Geny Anthony
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  4. Nagesh Sardesai
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Contributions

DP and SC designed research on method development. DP, SC, GA, NS, OK, TM, TM, PS, RS contributed to experimental design, transformations, data collection, data analysis, and/or preparation of manuscript. LC performed molecular analysis and contributed to preparation of manuscript. DG, BB, NS were responsible for plant care, T1 seed production and heritability studies. KS, RS monitored the progress and reviewed the manuscript.

Corresponding author

Correspondence to Nagesh Sardesai.

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Conflict of interest

GA, NS, TM, LC, DG, PS and KS are employed by Corteva Agriscience. TM, BB, NS and RS are employed by Inari, AgReliant Genetics, Syngenta, and AgBiome, Inc, respectively. DP, SC and OK declare that they have no conflict of interest.

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Pareddy, D., Chennareddy, S., Anthony, G. et al. Improved soybean transformation for efficient and high throughput transgenic production. Transgenic Res 29, 267–281 (2020). https://doi.org/10.1007/s11248-020-00198-8

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