title: Transient silencing of the grapevine gene VvPGIP1 by agroinfiltration with a construct for RNA interference.
abstract: Grapevine is an economically important crop, and the recent completion of its genome makes it possible to study the function of specific genes through reverse genetics. However, the analysis of gene function by RNA interference (RNAi) in grapevine is difficult, because the generation of stable transgenic plants has low efficiency and is time consuming. Recently, transient expression of genes in grapevine leaves has been obtained by Agrobacterium tumefaciens infiltration (agroinfiltration). We therefore tested the possibility to silence grapevine genes by agroinfiltration of RNAi constructs. A construct to express a double strand RNA (dsRNA) corresponding to the defense-related gene VvPGIP1, encoding a polygalacturonase-inhibiting protein (PGIP), was obtained and transiently expressed by agroinfiltration in leaves of grapevine plants grown in vitro. Expression of VvPGIP1 and accumulation of PGIP activity were strongly induced by infiltration with control bacteria, but not with bacteria carrying the dsRNA construct, indicating that the gene was efficiently silenced. In contrast, expression of another defense-related gene, VST1, encoding a stilbene synthase, was unaffected by the dsRNA construct. We have therefore demonstrated the possibility of transient down-regulation of grapevine genes by agroinfiltration of constructs for the expression of dsRNA. This system can be employed to evaluate the effectiveness of constructs that can be subsequently used to generate stable RNAi transgenic plants.
authors.affiliation: CRA-VIT Centro di ricerca per la viticoltura, Conegliano, TV, Italy.
authors.forename: Nadia
authors.initials: N
authors.lastname: Bertazzon
authors.forename: Alessandro
authors.initials: A
authors.lastname: Raiola
authors.forename: Carla
authors.initials: C
authors.lastname: Castiglioni
authors.forename: Massimo
authors.initials: M
authors.lastname: Gardiman
authors.forename: Elisa
authors.initials: E
authors.lastname: Angelini
authors.forename: Michele
authors.initials: M
authors.lastname: Borgo
authors.forename: Simone
authors.initials: S
authors.lastname: Ferrari
mesh_terms: D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism; D016960:Agrobacterium tumefaciens / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D018515:Plant Leaves / Q000235:genetics / Q000382:microbiology; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D012330:RNA, Double-Stranded; D059386:Reverse Genetics; D027843:Vitis / Q000235:genetics* / Q000378:metabolism / Q000382:microbiology
chemical_list: C081479:PGIP protein, plant; D010940:Plant Proteins; D012330:RNA, Double-Stranded; D000217:Acyltransferases; C022674:stilbene synthase