AgrobacTransV1, Main, Exploration, bibRecord, 000659

Agrobacterium-mediated transient gene expression and silencing: a rapid tool for functional gene assay in potato.

Identifieur interne : 000659 ( Main/Exploration ); précédent : 000658; suivant : 000660

Agrobacterium-mediated transient gene expression and silencing: a rapid tool for functional gene assay in potato.

Auteurs : Pudota B. Bhaskar [États-Unis] ; Muthusubramanian Venkateshwaran ; Lei Wu ; Jean-Michel Ané ; Jiming Jiang

Source :

PloS one [ 1932-6203 ] ; 2009.

RBID : pubmed:19503835

Descripteurs français

English descriptors

KwdEn :
- Gene Expression Regulation (MeSH), Gene Expression Regulation, Plant (MeSH), Gene Silencing (MeSH), Genetic Techniques (MeSH), Genome, Plant (MeSH), Genotype (MeSH), Green Fluorescent Proteins (metabolism), Phytophthora infestans (metabolism), Plant Diseases (genetics), Plant Diseases (microbiology), Plants, Genetically Modified (MeSH), Plasmids (metabolism), RNA Interference (MeSH), Rhizobium (genetics), Solanum tuberosum (genetics), Solanum tuberosum (microbiology).
MESH :
- chemical , metabolism : Green Fluorescent Proteins.
- genetics : Plant Diseases, Rhizobium, Solanum tuberosum.
- metabolism : Phytophthora infestans, Plasmids.
- microbiology : Plant Diseases, Solanum tuberosum.
- Gene Expression Regulation, Gene Expression Regulation, Plant, Gene Silencing, Genetic Techniques, Genome, Plant, Genotype, Plants, Genetically Modified, RNA Interference.

Abstract

Potato is the third most important food crop worldwide. However, genetic and genomic research of potato has lagged behind other major crops due to the autopolyploidy and highly heterozygous nature associated with the potato genome. Reliable and technically undemanding techniques are not available for functional gene assays in potato. Here we report the development of a transient gene expression and silencing system in potato. Gene expression or RNAi-based gene silencing constructs were delivered into potato leaf cells using Agrobacterium-mediated infiltration. Agroinfiltration of various gene constructs consistently resulted in potato cell transformation and spread of the transgenic cells around infiltration zones. The efficiency of agroinfiltration was affected by potato genotypes, concentration of Agrobacterium, and plant growth conditions. We demonstrated that the agroinfiltration-based transient gene expression can be used to detect potato proteins in sub-cellular compartments in living cells. We established a double agroinfiltration procedure that allows to test whether a specific gene is associated with potato late blight resistance pathway mediated by the resistance gene RB. This procedure provides a powerful approach for high throughput functional assay for a large number of candidate genes in potato late blight resistance.

DOI: 10.1371/journal.pone.0005812
PubMed: 19503835
PubMed Central: PMC2686102

Affiliations:

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Le document en format XML

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<front><div type="abstract" xml:lang="en">Potato is the third most important food crop worldwide. However, genetic and genomic research of potato has lagged behind other major crops due to the autopolyploidy and highly heterozygous nature associated with the potato genome. Reliable and technically undemanding techniques are not available for functional gene assays in potato. Here we report the development of a transient gene expression and silencing system in potato. Gene expression or RNAi-based gene silencing constructs were delivered into potato leaf cells using Agrobacterium-mediated infiltration. Agroinfiltration of various gene constructs consistently resulted in potato cell transformation and spread of the transgenic cells around infiltration zones. The efficiency of agroinfiltration was affected by potato genotypes, concentration of Agrobacterium, and plant growth conditions. We demonstrated that the agroinfiltration-based transient gene expression can be used to detect potato proteins in sub-cellular compartments in living cells. We established a double agroinfiltration procedure that allows to test whether a specific gene is associated with potato late blight resistance pathway mediated by the resistance gene RB. This procedure provides a powerful approach for high throughput functional assay for a large number of candidate genes in potato late blight resistance.</div>
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<ArticleIdList><ArticleId IdType="pubmed">15077675</ArticleId>
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</Reference>
<Reference><Citation>Plant Mol Biol. 2005 Nov;59(4):647-61</Citation>
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<affiliations><list><country><li>États-Unis</li>
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<tree><noCountry><name sortKey="Ane, Jean Michel" sort="Ane, Jean Michel" uniqKey="Ane J" first="Jean-Michel" last="Ané">Jean-Michel Ané</name>
<name sortKey="Jiang, Jiming" sort="Jiang, Jiming" uniqKey="Jiang J" first="Jiming" last="Jiang">Jiming Jiang</name>
<name sortKey="Venkateshwaran, Muthusubramanian" sort="Venkateshwaran, Muthusubramanian" uniqKey="Venkateshwaran M" first="Muthusubramanian" last="Venkateshwaran">Muthusubramanian Venkateshwaran</name>
<name sortKey="Wu, Lei" sort="Wu, Lei" uniqKey="Wu L" first="Lei" last="Wu">Lei Wu</name>
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<country name="États-Unis"><region name="Wisconsin"><name sortKey="Bhaskar, Pudota B" sort="Bhaskar, Pudota B" uniqKey="Bhaskar P" first="Pudota B" last="Bhaskar">Pudota B. Bhaskar</name>
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Agrobacterium-mediated transient gene expression and silencing: a rapid tool for functional gene assay in potato.

Agrobacterium-mediated transient gene expression and silencing: a rapid tool for functional gene assay in potato.

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