ArticleThe Plant journal : for cell and molecular biology2014
Direct visualization of Agrobacterium-delivered VirE2 in recipient cells.
Article in The Plant journal : for cell and molecular biology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed.
- Myosin VIII and XI isoforms interact with the Agrobacterium VirE2 protein and facilitate its transport from the plasma membrane to the perinuclear region during plant transformation.Plant communications · 2026Article
- Using the Superfolder GFP (sfGFP) System to Study Plant Peroxisomal Protein Import.Methods in molecular biology (Clifton, N.J.) · 2023Article
- A tau class GST, OsGSTU5, interacts with VirE2 and modulates the Agrobacterium-mediated transformation in rice.Plant cell reports · 2022Article
- Genetic factors governing bacterial virulence and host plant susceptibility duringAdvances in genetics · 2022Review
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- Single Molecule Imaging of T-DNA Intermediates FollowingInternational journal of molecular sciences · 2019Article
- Pathways of DNA Transfer to Plants fromAnnual review of phytopathology · 2019Article
- Development and Applications of Superfolder and Split Fluorescent Protein Detection Systems in Biology.International journal of molecular sciences · 2019Review
- Split Green Fluorescent Protein System to Visualize Effectors Delivered from Bacteria During Infection.Journal of visualized experiments : JoVE · 2018Article
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- Spatiotemporal Monitoring ofThe Plant cell · 2017Article
- Direct and Indirect Visualization of Bacterial Effector Delivery into Diverse Plant Cell Types during Infection.The Plant cell · 2017Article
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- A self-perpetuating repressive state of a viral replication protein blocks superinfection by the same virus.PLoS pathogens · 2017Article
- Agrobacterium-mediated plant transformation: biology and applications.The arabidopsis book · 2017Article
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Authors and funding
5 authors.
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No grant is acknowledged in the PubMed record.
Abstract
Agrobacterium tumefaciens is a natural genetic engineer widely used to deliver DNA into various recipients, including plant, yeast and fungal cells. The bacterium can transfer single-stranded DNA molecules (T-DNAs) and bacterial virulence proteins, including VirE2. However, neither the DNA nor the protein molecules have ever been directly visualized after the delivery. In this report, we adopted a split-GFP approach: the small GFP fragment (GFP11) was inserted into VirE2 at a permissive site to create the VirE2-GFP11 fusion, which was expressed in A. tumefaciens; and the large fragment (GFP1-10) was expressed in recipient cells. Upon delivery of VirE2-GFP11 into the recipient cells, GFP fluorescence signals were visualized. VirE2-GFP11 was functional like VirE2; the GFP fusion movement could indicate the trafficking of Agrobacterium-delivered VirE2. As the natural host, all plant cells seen under a microscope received the VirE2 protein in a leaf-infiltration assay; most of VirE2 moved at a speed of 1.3-3.1 μm sec⁻¹ in a nearly linear direction, suggesting an active trafficking process. Inside plant cells, VirE2-GFP formed filamentous structures of different lengths, even in the absence of T-DNA. As a non-natural host recipient, 51% of yeast cells received VirE2, which did not move inside yeast. All plant cells seen under a microscope transiently expressed the Agrobacterium-delivered transgene, but only 0.2% yeast cells expressed the transgene. This indicates that Agrobacterium is a more efficient vector for protein delivery than T-DNA transformation for a non-natural host recipient: VirE2 trafficking is a limiting factor for the genetic transformation of a non-natural host recipient. The split-GFP approach could enable the real-time visualization of VirE2 trafficking inside recipient cells.
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