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A procedure for the transient expression of genes by agroinfiltration above the permissive threshold to study temperature-sensitive processes in plant–pathogen interactions

AutorDel Toro, Francisco ; Tenllado, Francisco ; Chung, Bong-Nam; Canto, Tomás
Palabras claveAgroinfiltration
Plant virology
Plant–virus interactions
Transient expression
Fecha de publicación14-abr-2014
EditorJohn Wiley & Sons
CitaciónMolecular Plant Pathology
ResumenLocalized expression of genes in plants from T-DNAs delivered into plant cells by Agrobacterium tumefaciens is an important tool in plant research. The technique, known as agroinfiltration, provides fast, efficient ways to transiently express or silence a desired gene without resorting to the time-consuming, challenging stable transformation of the host, use of less efficient means of delivery, such as bombardment, or use of viral vectors, which multiply and spread within the host causing themselves physiological alterations. A drawback of the agroinfiltration technique is its temperature dependency: early works had shown that temperatures above 29 °C were non-permissive to tumor induction by the bacterium as result of failure in pilus formation. However, research in plant sciences has interest in studying processes at those temperatures, above the 25 °C experimental standard, common to many host-environment and host-pathogen interactions in nature, and agroinfiltration would be an excellent tool for this purpose. Here we have measured the efficiency of agroinfiltration for expressing reporter genes in plants from T-DNAs at non-permissive 30 °C, either transiently or as part of viral amplicons, and envisaged procedures that allow and optimize its use for gene expression at that temperature. We have applied this technical advance to assess the performance at 30 °C of two viral suppressors of silencing in agropatch assays: Potato virus Y HCPro and Cucumber mosaic virus 2b protein, and within the context of infection by a PVX vector, indirectly assessed also their effect on the overall response of the host Nicotiana benthamiana to the virus.
Descripción24 p.-1 tab.-3 fig.
Versión del editorhttp://dx.doi.org/10.1111/mpp.12136
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