Turnip mosaic virus (TuMV) constitutes one of the primary diseases affecting Brassica rapa, severely impacting its production and resulting in crop failures in various regions worldwide. Recent research has demonstrated the significance of plant translation initiation factors, specifically the eIF4E and eIF4G family genes, as essential recessive disease resistance genes. In our study, we conducted evolutionary and gene expression studies, leading us to identify eIF(iso)4E.c as a potential TuMV-resistant gene. Leveraging CRISPR/Cas9 technology, we obtained mutant B. rapa plants with edited eIF(iso)4E.c gene. We confirmed eIF(iso)4E.c confers resistance against TuMV through phenotypic observations and virus content evaluations. Furthermore, we employed ribosome profiling assays on eif(iso)4e.c mutant seedlings to unravel the translation landscape in response to TuMV. Interestingly, we observed a moderate correlation between the fold changes in gene expression at the transcriptional and translational levels (R2 = 0.729). Comparative analysis of ribosome profiling and RNA-seq data revealed that plant–pathogen interaction, and MAPK signaling pathway–plant pathways were involved in eIF(iso)4E.c-mediated TuMV resistance. Further analysis revealed that sequence features, coding sequence length, and normalized minimal free energy, influenced the translation efficiency of genes. Our study highlights that the loss of eIF(iso)4E.c can result in a highly intricate translation mechanism, acting synergistically with transcription to confer resistance against TuMV.
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Brassica rapa L. is cultivated globally and consumed in many areas worldwide. Using the transgenic Agrobacterium-mediated transformation method, which is a reproducible and efficient technique, genes can be transferred into various B. rapa species. This review summarizes the processes involved in Agrobacterium-mediated transformation of B. rapa, including surface seed sterilization, co-cultivation with A. tumefaciens, induction of callus/shoot/root formation, and confirmation of transgenic plants. In addition, factors such as the Agrobacterium strain, plant genotype, explant age, transformation efficiency of the hybrid or inbred line, and the concentrations of N6-benzyl amino purine and naphthalene acetic acid, are discussed. And this review shows clearly how to do it, what to do, and what not to do in the transgenic Agrobacterium-mediated in Brassica rapa. The information presented here lays the foundation for a simple and efficient method that resolves existing problems and improves overall transgenic B. rapa production, thereby benefiting both basic and applied research.
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