title: Virus induced gene silencing (VIGS) for functional analysis of wheat genes involved in Zymoseptoria tritici susceptibility and resistance.
abstract: Virus-induced gene silencing (VIGS) has emerged as a powerful reverse genetic technology in plants supplementary to stable transgenic RNAi and, in certain species, as a viable alternative approach for gene functional analysis. The RNA virus Barley stripe mosaic virus (BSMV) was developed as a VIGS vector in the early 2000s and since then it has been used to study the function of wheat genes. Several variants of BSMV vectors are available, with some requiring in vitro transcription of infectious viral RNA, while others rely on in planta production of viral RNA from DNA-based vectors delivered to plant cells either by particle bombardment or Agrobacterium tumefaciens. We adapted the latest generation of binary BSMV VIGS vectors for the identification and study of wheat genes of interest involved in interactions with Zymoseptoria tritici and here present detailed and the most up-to-date protocols.
authors.affiliation: Plant Biology and Crop Science Department, Rothamsted Research, Harpenden AL5 2JQ, United Kingdom.
authors.forename: Wing-Sham
authors.initials: WS
authors.lastname: Lee
authors.affiliation: Plant Biology and Crop Science Department, Rothamsted Research, Harpenden AL5 2JQ, United Kingdom.
authors.forename: Jason J
authors.initials: JJ
authors.lastname: Rudd
authors.affiliation: Plant Biology and Crop Science Department, Rothamsted Research, Harpenden AL5 2JQ, United Kingdom. Electronic address: kostya.kanyuka@rothamsted.ac.uk.
authors.forename: Kostya
authors.initials: K
authors.lastname: Kanyuka
mesh_terms: D001203:Ascomycota / Q000254:growth & development / Q000276:immunology; D060467:Disease Resistance; D004198:Disease Susceptibility; D055785:Gene Knockdown Techniques / Q000379:methods*; D020868:Gene Silencing; D017343:Genes, Plant; D005822:Genetic Vectors; D010935:Plant Diseases / Q000235:genetics* / Q000382:microbiology; D010942:Plant Viruses / Q000235:genetics*; D012328:RNA Viruses / Q000235:genetics; D059386:Reverse Genetics / Q000379:methods; D014170:Transformation, Genetic; D014908:Triticum / Q000235:genetics* / Q000382:microbiology
keywords: Functional genomics; Mycosphaerella graminicola; RNAi; Septoria tritici; Triticum aestivum; Virus induced gene silencing (VIGS); Wheat