title: Virus-induced gene silencing for comparative functional studies in Gladiolus hybridus.
abstract: Functional analysis of genes in gladiolus has previously been impractical due to the lack of an efficient stable genetic transformation method. However, virus-induced gene silencing (VIGS) is effective in some plants which are difficult to transform through other methods. Although the Tobacco rattle virus (TRV)-based VIGS system has been developed and used for verifying gene functions in diverse plants, an appropriate TRV-VIGS approach for gladiolus has not been established yet. In this report we describe the first use of the TRV-VIGS system for gene silencing in gladiolus. Vacuum infiltration of cormels and young plants with the GhPDS-VIGS vector effectively down-regulated the PHYTOENE DESATURASE ortholog GhPDS gene and also resulted in various degrees of photobleaching in Gladiolus hybridus. The reduction in GhPDS expression was tested after TRV-based vector infection using real-time RT-PCR. In addition, the progress of TRV infection was detected by fluorescence visualization using a pTRV2: CP-GFP vector. In conclusion, the TRV-mediated VIGS described here will be an effective gene function analysis mechanism in gladiolus.
authors.affiliation: Department of Ornamental Horticulture and Landscape Architecture, China Agricultural University, Yuan Mingyuan Western Road 2#, Beijing, 100193, China, xionghui.zhong2012@gmail.com.
authors.forename: Xionghui
authors.initials: X
authors.lastname: Zhong
authors.forename: Xue
authors.initials: X
authors.lastname: Yuan
authors.forename: Ze
authors.initials: Z
authors.lastname: Wu
authors.forename: Muhammad Ali
authors.initials: MA
authors.lastname: Khan
authors.forename: Jin
authors.initials: J
authors.lastname: Chen
authors.forename: Xiaoxin
authors.initials: X
authors.lastname: Li
authors.forename: Benhe
authors.initials: B
authors.lastname: Gong
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhao
authors.forename: Jian
authors.initials: J
authors.lastname: Wu
authors.forename: Chenyu
authors.initials: C
authors.lastname: Wu
authors.forename: Mingfang
authors.initials: M
authors.lastname: Yi
mesh_terms: D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D027621:Iridaceae / Q000166:cytology / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D010641:Phenotype; D018515:Plant Leaves / Q000166:cytology / Q000235:genetics / Q000378:metabolism / Q000821:virology; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000166:cytology / Q000235:genetics / Q000378:metabolism / Q000821:virology; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins; D036226:Seedlings / Q000166:cytology / Q000235:genetics / Q000378:metabolism / Q000821:virology; D017422:Sequence Analysis, DNA
chemical_list: D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D010088:Oxidoreductases; C022398:phytoene dehydrogenase