The use of protoplasts to study innate immune responses.

@article{He2007TheUO,
  title={The use of protoplasts to study innate immune responses.},
  author={Ping He and Libo Shan and Jen Sheen},
  journal={Methods in molecular biology},
  year={2007},
  volume={354},
  pages={
          1-9
        }
}
The use of plant protoplast transient expression system has facilitated the discovery and dissection of many signal transduction pathways in response to hormones, metabolites, and stresses. Recently, Arabidopsis protoplasts also have been used successfully to study plant innate immune responses triggered by pathogen-derived elicitors. Here, we describe the detailed protocols for studying innate immune responses, including cell death and early defense gene regulation activated by two types of… 

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References

SHOWING 1-10 OF 21 REFERENCES
Signal transduction in maize and Arabidopsis mesophyll protoplasts.
  • J. Sheen
  • Environmental Science, Biology
    Plant physiology
  • 2001
TLDR
The development of defined protoplast transient expression systems for high-throughput screening and systematic characterization of gene functions has greatly contributed to elucidating plant signal transduction pathways, in combination with genetic, genomic, and transgenic approaches.
Understanding the functions of plant disease resistance proteins.
Many disease resistance (R) proteins of plants detect the presence of disease-causing bacteria, viruses, or fungi by recognizing specific pathogen effector molecules that are produced during the
Signal transduction in the plant immune response.
Direct delivery of bacterial avirulence proteins into resistant Arabidopsis protoplasts leads to hypersensitive cell death.
TLDR
It is demonstrated that E. coli-produced AvrRpt2 and AvrB proteins must enter the plant cell to be recognized by R gene-mediated mechanisms, and that a method based on a carrier peptide can be used to introduce proteins into plant cells.
MAP kinase signalling cascade in Arabidopsis innate immunity
TLDR
An Arabidopsis thaliana leaf cell system based on the induction of early-defence gene transcription by flagellin, a highly conserved component of bacterial flagella that functions as a PAMP in plants and mammals is developed, suggesting that signalling events initiated by diverse pathogens converge into a conserved MAPK cascade.
Disabling surveillance: bacterial type III secretion system effectors that suppress innate immunity
TLDR
Type III effectors can suppress signal transduction pathways activated by PRR surveillance systems, which will reveal much about bacterial pathogenicity and the innate immune system in plants and animals.
Plant pathogens and integrated defence responses to infection
TLDR
The current knowledge of recognition-dependent disease resistance in plants is reviewed, and a few crucial concepts are included to compare and contrast plant innate immunity with that more commonly associated with animals.
FLS2: an LRR receptor-like kinase involved in the perception of the bacterial elicitor flagellin in Arabidopsis.
Plants have a sensitive perception system for the most conserved domain of bacterial flagellin.
TLDR
It is demonstrated that plants have a highly sensitive chemoperception system for eubacterial flagellins, specifically targeted to the most highly conserved domain within its N terminus.
...
1
2
3
...