ArticlePlant methods2024
A high-efficiency transient expression system mediated by Agrobacterium tumefaciens in Spinacia oleracea leaves.
Article in Plant methods, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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5 citing papers in PubMed.
- Establishment of Transient Expression and Hairy Root Induction Systems inPlants (Basel, Switzerland) · 2026Article
- PpBRC1 negatively regulates branching via modulating GA signal transduction gene PpGID1b in peach (Prunus persica).Plant cell reports · 2026Article
- Advancing the frontier of plant-based therapeutics: critical innovations in molecular farming and bioprocess Integration.Biotechnology letters · 2026Review
- Article
- Transgene-free genome editing in citrus and poplar trees using positive and negative selection markers.Plant cell reports · 2025Article
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7 authors.
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Abstract
backgroundOptimization of a highly efficient transient expression system is critical for the study of gene function, particularly in those plants in which stable transformation methods are not widely available. Agrobacterium tumefaciens‑mediated transient transformation is a simple and low-cost method that has been developed and applied to a wide variety of plant species. However, the transient expression in spinach (Spinacia oleracea L.) is still not reported.
resultsWe developed a transient expression system in spinach leaves of the Sp75 and Sp73 varieties. Several factors influencing the transformation efficiency were optimized such as Agrobacterium strain, spinach seedling stage, leaf position, and the expression time after injection. Agrobacterium strain GV3101 (pSoup-p19) was more efficient than AGL1 in expressing recombinant protein in spinach leaves. In general, Sp75 leaves were more suitable than Sp73 leaves, regardless of grow stage. At four-leaf stage, higher intensity and efficiency of transient expression were observed in group 1 (G1) of Sp75 at 53 h after injection (HAI) and in G1 of Sp73 at 64 HAI. At six-leaf stage of Sp75, group 3 (G3) at 72 HAI were the most effective condition for transient expression. Using the optimized expression system, we detected the subcellular localization of a transcriptional co-activator SoMBF1c and a NADPH oxidase SoRbohF. We also detected the interaction of the protein kinase SoCRK10 and the NADPH oxidase SoRbohB.
conclusionThis study established a method of highly efficient transient expression mediated by Agrobacterium in spinach leaves. The transient expression system will facilitate the analysis of gene function and lay a solid foundation for molecular design breeding of spinach.
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