数字农科院2.0

An efficient soybean transformation protocol for use with elite lines

文献类型： 外文期刊

作者： Wang, Ying;Li, Zhan;Chen, Xiaorui;Gu, Yongzhe;Zhang, Lijuan;Qiu, Lijuan

作者机构：

关键词： Soybean;Agrobacterium-mediated transformation;Elite line;Glyphosate selection

期刊名称： PLANT CELL TISSUE AND ORGAN CULTURE

ISSN： 0167-6857

年卷期： 2023 年

页码：

收录情况： SCIE（2023版） ; ; EI（2023版）

摘要： Key message High transformation rates, even in elite soybean varieties, can be achieved via this highly efficient method for Agrobacterium-mediated transformation using a seed-based explant type, with suitable media for each stage, selectable markers, and desiccation treatment during co-culture. The transformation frequencies of soybean (Glycine max) are low (< 10%), especially when glyphosate is used as the selection agent. Here, we describe a highly efficient soybean transformation method based on the use of mature seed halves containing mature embryos with the cotyledonary leaves removed as explants. Suitable media composition and culture condition lead to high transformation frequency. Except explant type and media, we used a different selectable marker and an improved desiccation co-culture treatment to improve transformation efficiency. Co-culture on a single piece of filter paper moistened with 1 mL liquid medium to induce desiccation significantly improved transient expression level compared to co-culture on solid medium, which is commonly used in plant transformation. The use of the glyphosate resistance gene CP4-EPSPS as a selectable marker led to a higher regeneration rate (24.7%) and transformation frequency (22.9%) compared to the glyphosate resistance gene CG10-EPSPS (13.8% and 13.6%, respectively). This protocol resulted in transformation frequencies of up to 31.7% when the soybean variety 'Jack' was used as the recipient. Using this system, 92.5% of regenerated shoots were transgenic. Moreover, the transgene in approximately 89% of T-0 plants was inherited by T-1 progeny based on quantitative RT-PCR analysis. Our method was successfully applied to 15 elite soybean lines, with transformation frequencies of 3-35.8%; half of these lines showed high transformation frequencies. This protocol should facilitate basic research and enhance genetic engineering of soybean.

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