This study developed an efficient virus-induced gene silencing (VIGS) system using Tobacco ring spot virus (TRSV) as a vector in cucumber (Cucumis sativus). TRSV is currently the most efficient VIGS vector. The key steps are designing the viral genome to insert fragments of the host target gene to be silenced, infecting the plant host, and silencing the target gene as part of the plant's antiviral defense response. For TRSV-based VIGS in cucumber, Agrobacterium containing modified pTRSV1 and pTRSV2 vectors with the plant target gene fragment are mixed and infiltrated into lower leaves. After 2-3 weeks, the target

1. Development of an efficient VIGS system using Tobacco ring spot virus vector in Cucumis sativus
Mesele Tilahun1, 2 and Jae Son Moon 1, 2
1Korea Research Institute of Bioscience and Biotechnology, 125 Gwahak-ro, Yuseong-gu,
Daejeon 34114, South Korea
2University of Science and Technology (UST), Yuseong, Daejeon 34113, South Korea
Virus-induced gene silencing (VIGS) is one of the methods of reverse genetics for gene function analysis and a newly
evolved technique of gene transcript suppression for plant gene function characterization. The family Cucurbitaceae
has abundant flower and plant sex types, and the regulation of sex determination can directly influence their yield and
quality Tobacco ring spot virus (TRSV)-based VIGS vector is the most efficient VIGS vector so far. Here, this study
describe a rapid and high efficient TRSV-based VIGS method for knocking down sex determinant gene of in Cucumis
sativus. The key steps in VIGS include: designing viral genomes to insert fragments of host genes that are targeted to
be silenced into the relevant viral vector, infecting the suitable plant hosts and silencing the target genes as part of the
plant's defense mechanism preventing virus infection. For TRSV-based VIGS, Agrobacterium culture containing
pTRSV1 and Agrobacterium culture containing pTRSV2 with plant target gene fragment are mixed and infiltrated into
the lower leaves of plant. After 2-3 weeks post infiltration, plant target gene will be silenced.
Hypothesis : Ethylene influences the sex-controlling
genes (ACS and ACO gene) in cucumber.
Virus-induced gene silencing (VIGS) is one of the methods of
reverse genetics for gene function analysis and a newly evolved
technique of gene transcript suppression for plant gene function
characterization. VIGS is a quick, effective and precise method
for transient gene silencing.
Overview of Experiments
fig.2 major steps of VIGS experiments
fig.3. cloning and infiltration
fig.4 expected result of silenced vs. non
silenced transcriptome expression
Fig .1 Molecular mechanism of VIGS
short interfering RNA
RNA-induced Silencing Complex
Including Argonaute
degraded
Repetitive, aberrant, viral