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A robust high-throughput functional screening assay for plant pathogen effectors using the TMV-GFP vector.

Peng Cao1, Haotian Shi1,2, Shuangxi Zhang1

  • 1College of Life Sciences, Shaanxi Normal University, Xi'an, 710119, China.

The Plant Journal : for Cell and Molecular Biology
|April 22, 2024
PubMed
Summary

Researchers developed a novel screening system using tobacco mosaic virus (TMV)-GFP to rapidly identify plant immune-interfering pathogen effectors. This efficient method aids in understanding plant diseases and developing crop protection strategies.

Keywords:
TMV‐GFPavirulenceeffectorfunctional screeningtechnical advancevirulence

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Area of Science:

  • Plant Pathology
  • Molecular Plant-Microbe Interactions
  • Biotechnology

Background:

  • Understanding phytopathogen effectors is key to deciphering pathogenicity mechanisms and enhancing plant disease resistance.
  • Functional characterization of numerous predicted effectors is a critical research focus in plant-pathogen interactions.

Purpose of the Study:

  • To design and validate a novel, rapid screening system for identifying plant immune-interfering pathogen effectors.
  • To evaluate the efficiency and broad applicability of the developed system for effector screening.

Main Methods:

  • Development of a screening system combining tobacco mosaic virus (TMV)-GFP vector and Agrobacterium-mediated transient expression in Nicotiana benthamiana.
  • Evaluation of the system using known effectors from Ralstonia solanacearum.
  • Screening for novel virulence effectors and assessment of cytoplasmic and apoplastic effector identification capabilities.

Main Results:

  • The TMV-GFP system accurately and efficiently identified known effectors, including a novel virulence effector, RipS1.
  • Consistent results were obtained for previously characterized cytoplasmic effectors from diverse pathogens.
  • The system demonstrated effectiveness in identifying apoplastic effectors.

Conclusions:

  • The TMV-GFP screening system offers a rapid, efficient, and broadly applicable method for identifying pathogen effectors that interfere with plant immunity.
  • Its ease of operation, time-saving nature, and low technical requirements make it suitable for high-throughput screening.