Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana

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...

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Main Authors:	Du, Juan, Rietman, Hendrik, Vleeshouwers, Vivianne G. A. A.
Format:	Online
Language:	English
Published:	MyJove Corporation 2014
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4063549/

id	pubmed-4063549
recordtype	oai_dc
spelling	pubmed-40635492014-06-25 Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana Du, Juan Rietman, Hendrik Vleeshouwers, Vivianne G. A. A. Plant Biology 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. MyJove Corporation 2014-01-03 /pmc/articles/PMC4063549/ /pubmed/24430891 http://dx.doi.org/10.3791/50971 Text en Copyright © 2014, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Du, Juan Rietman, Hendrik Vleeshouwers, Vivianne G. A. A.
spellingShingle	Du, Juan Rietman, Hendrik Vleeshouwers, Vivianne G. A. A. Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana
author_facet	Du, Juan Rietman, Hendrik Vleeshouwers, Vivianne G. A. A.
author_sort	Du, Juan
title	Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana
title_short	Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana
title_full	Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana
title_fullStr	Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana
title_full_unstemmed	Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana
title_sort	agroinfiltration and pvx agroinfection in potato and nicotiana benthamiana
description	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.
publisher	MyJove Corporation
publishDate	2014
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4063549/
_version_	1612103628492898304

Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana

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