The role of a soybean 14-3-3 gene (Glyma05g29080) on white mold resistance and nodulation investigations using CRISPR-Cas9 editing and RNA silencing

Authors: ZHANG, YUNFANG - Agriculture And Agri-Food Canada; BLAHUT-BEATTY, LAUREEN - Agriculture And Agri-Food Canada; ZHENG, SUQIN - Agriculture And Agri-Food Canada; Clough, Steven; SIMMONDS, DAINA - Agriculture And Agri-Food Canada

Submitted to: Molecular Plant-Microbe Interactions
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/25/2022
Publication Date: 11/23/2022
Citation: Zhang, Y., Blahut-Beatty, L., Zheng, S., Clough, S.J., Simmonds, D.H. 2022. The role of a soybean 14-3-3 gene (Glyma05g29080) on white mold resistance and nodulation investigations using CRISPR-Cas9 editing and RNA silencing. Molecular Plant-Microbe Interactions. 36(3):159-164. https://doi.org/10.1094/MPMI-07-22-0157-R.
DOI: https://doi.org/10.1094/MPMI-07-22-0157-R

Interpretive Summary: It can be a challenge to determine the precise role of an individual gene. To determine what a given gene might be doing for a plant, one can mutate that gene and observe how the mutant plant behaves. In plants, it is now possible to specifically mutate a gene of interest using the technology called CRISPR-Cas9. We used this technique to specifically mutate a 14-3-3 gene which encodes for a class of proteins known to be involved in signal transduction within plants and animals. We found that plants that have had this specific 14-3-3 gene mutated had decreased nodulation and were more susceptible to disease development after inoculation with the pathogen Sclerotinia sclerotiorum. Therefore, we provide strong evidence that this specific 14-3-3 is involved in healthy nodulation as well as plays a beneficial role in reducing the levels of disease development. This work will be of interest to the researchers in the fields of signal transduction and plant-microbe interactions.

Technical Abstract: The role of soybean 14-3-3 gene (Glyma05g29080) in defense against white mold and in nodulation was investigated by loss-of-gene-function with CRISPR-Cas9 editing and RNAi silencing. Particle bombardment was used to introduce the CRISPR expression cassette to target the soybean 14-3-3 gene and an RNAi construct to silence gene transcription. Transmission of the edited 14-3-3 gene and the RNAi construct was confirmed in their respective progeny. The recovered transgenic plants and their progeny were significantly more susceptible to S. sclerotiorum infection and showed a significant reduction in nodulation, thus confirming the role of the 14-3-3 (Glyma05g29080) gene in both nodulation and defense.