title: Virus-induced gene silencing in Medicago truncatula and Lathyrus odorata.
abstract: Virus-induced gene silencing (VIGS) has become an important reverse genetics tool for functional genomics. VIGS vectors based on Pea early browning virus (PEBV, genus Tobravirus) and Bean pod mottle virus (genus Comovirus) are available for the legume species Pisum sativum and Glycine max, respectively. With the aim of extending the application of the PEBV VIGS vector to other legumes, we examined susceptibility of 99 accessions representing 24 legume species including 21 accessions of Medicago truncatula and 38 accessions Lotus japonicus. Infectivity of PEBV was tested by agro-inoculation with a vector carrying the complete beta-glucuronidase (GUS) coding sequence. In situ histochemical staining analysis indicated that 4 of 21 M. truncatula and three of three Lathyrus odorata accessions were infected systemically by GUS tagged PEBV, while none of 38 L. japonicus accessions displayed GUS staining of either inoculated or uninoculated leaves. Agro-inoculation of plants representing PEBV-GUS susceptible M. truncatula and L. odorata accessions with PEBV carrying a fragment of Phytoene desaturase (PDS) resulted in development of a bleaching phenotype suggesting a down-regulation of PDS expression. In M. truncatula this was supported by quantification of PDS mRNA levels by real-time PCR.
authors.affiliation: Department of Genetics and Biotechnology, Faculty of Agricultural Sciences, University of Aarhus, Thorvaldsensvej 40, DK-1871 Frederiksberg C, Denmark. mette.groenlund@risoe.dk
authors.forename: Mette
authors.initials: M
authors.lastname: Grønlund
authors.forename: Gabriela
authors.initials: G
authors.lastname: Constantin
authors.forename: Elodie
authors.initials: E
authors.lastname: Piednoir
authors.forename: Jordan
authors.initials: J
authors.lastname: Kovacev
authors.forename: I Elisabeth
authors.initials: IE
authors.lastname: Johansen
authors.forename: Ole Søgaard
authors.initials: OS
authors.lastname: Lund
mesh_terms: D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D005822:Genetic Vectors; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism*; D027807:Lathyrus / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D000070116:Lotus / Q000235:genetics / Q000378:metabolism / Q000821:virology; D046913:Medicago truncatula / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism / Q000821:virology; D010942:Plant Viruses / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity; D030821:Plants, Genetically Modified; D012328:RNA Viruses / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*
chemical_list: D012333:RNA, Messenger; D005966:Glucuronidase