New genetic engineering techniques have advanced the field of plant molecular biology, and Agrobacterium-mediated transformation has enabled the discovery of numerous molecular and genetic functions. It has been widely used in many plants, including the economically important crop, soybean. Large-scale genetic analyses are needed to comprehend the molecular mechanisms that underlie the agronomic traits of soybean, and the generation of stable transgenic plants involves a lengthy and laborious process. Agrobacterium rhizogenes-mediated hairy root transformation is a quick and efficient method for investigations of root-specific processes and interactions. Generation of composite plants with transgenic roots and wild-type shoots allows for the study of the genetic mechanisms involved in root biology, such as the Bradyrhizobium-soybean interaction. Here, we provide an updated protocol for generating hairy soybean roots in as little as 18 days in a cost- and space-effective manner and demonstrate possible uses of composite plants with soybean nodulation assays and gene expression analysis as examples. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Soybean hairy root transformation. Basic Protocol 2: Soybean nodulation assay. Alternate Protocol: Soybean nodulation assay in germination pouches. Support Protocol: Bradyrhizobium japonicum culture preparation for inoculation. Basic Protocol 3: Histochemical GUS staining for promoter analysis.
CITATION STYLE
Song, J., Tóth, K., Montes-Luz, B., & Stacey, G. (2021). Soybean Hairy Root Transformation: A Rapid and Highly Efficient Method. Current Protocols, 1(7). https://doi.org/10.1002/cpz1.195
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