Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana

J. Du, H. Rietman, V.G.A.A. Vleeshouwers

Research output: Contribution to journalArticleAcademicpeer-review

26 Citations (Scopus)

Abstract

Agroinfiltration and PVX agroinfection are two efficient transient expression assays for functional analysis of candidate genes in plants. The most commonly used agent for agroinfiltration is Agrobacterium tumefaciens, a pathogen of many dicot plant species. This implies that agroinfiltration can be applied to many plant species. Here, we present our protocols and expected results when applying these methods to the potato (Solanum tuberosum), its related wild tuber-bearing Solanum species (Solanum section Petota) and the model plant Nicotiana benthamiana. In addition to functional analysis of single genes, such as resistance (R) or avirulence (Avr) genes, the agroinfiltration assay is very suitable for recapitulating the R-AVR interactions associated with specific host pathogen interactions by simply delivering R and Avr transgenes into the same cell. However, some plant genotypes can raise nonspecific defense responses to Agrobacterium, as we observed for example for several potato genotypes. Compared to agroinfiltration, detection of AVR activity with PVX agroinfection is more sensitive, more high-throughput in functional screens and less sensitive to nonspecific defense responses to Agrobacterium. However, nonspecific defense to PVX can occur and there is a risk to miss responses due to virus-induced extreme resistance. Despite such limitations, in our experience, agroinfiltration and PVX agroinfection are both suitable and complementary assays that can be used simultaneously to confirm each other's results.
Original languageEnglish
Article numbere50971
Number of pages7
JournalJournal of Visualized Experiments
Issue number83
DOIs
Publication statusPublished - 2014

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Solanum tuberosum
Tobacco
Assays
Functional analysis
Genes
Pathogens
Bearings (structural)
Solanum
Agrobacterium
Viruses
Genotype
Host-Pathogen Interactions
Agrobacterium tumefaciens
Throughput
Genetic Association Studies
Transgenes

Keywords

  • late blight resistance
  • mediated plant transformation
  • phytophthora-infestans
  • pathogen phytophthora
  • disease resistance
  • effector proteins
  • gene-expression
  • binary vector
  • agrobacterium
  • recognition

Cite this

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title = "Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana",
abstract = "Agroinfiltration and PVX agroinfection are two efficient transient expression assays for functional analysis of candidate genes in plants. The most commonly used agent for agroinfiltration is Agrobacterium tumefaciens, a pathogen of many dicot plant species. This implies that agroinfiltration can be applied to many plant species. Here, we present our protocols and expected results when applying these methods to the potato (Solanum tuberosum), its related wild tuber-bearing Solanum species (Solanum section Petota) and the model plant Nicotiana benthamiana. In addition to functional analysis of single genes, such as resistance (R) or avirulence (Avr) genes, the agroinfiltration assay is very suitable for recapitulating the R-AVR interactions associated with specific host pathogen interactions by simply delivering R and Avr transgenes into the same cell. However, some plant genotypes can raise nonspecific defense responses to Agrobacterium, as we observed for example for several potato genotypes. Compared to agroinfiltration, detection of AVR activity with PVX agroinfection is more sensitive, more high-throughput in functional screens and less sensitive to nonspecific defense responses to Agrobacterium. However, nonspecific defense to PVX can occur and there is a risk to miss responses due to virus-induced extreme resistance. Despite such limitations, in our experience, agroinfiltration and PVX agroinfection are both suitable and complementary assays that can be used simultaneously to confirm each other's results.",
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Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana. / Du, J.; Rietman, H.; Vleeshouwers, V.G.A.A.

In: Journal of Visualized Experiments, No. 83, e50971, 2014.

Research output: Contribution to journalArticleAcademicpeer-review

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