Abstract Detail

Evolutionary Developmental Biology (Evo-Devo)

Berger, Brent A. [1], Ricigliano, Vincent [1], Boyden, Geraldine [1], Howarth, Dianella [1].

Heterologous Gene-Silencing of PDS Using VIGS Across Three Related Species of Valerianaceae.

Virus-induced gene silencing (VIGS) is a homology-based RNA-silencing technique that utilizes the antiviral defense mechanism in plants to suppress gene expression. Viral vectors containing a fragment of the target gene are introduced into the host plant and lead to the production of double stranded RNA molecules that are recognized as foreign bodies and cleaved into small (21-23 nucleotide), double stranded fragments (siRNA; small interfering RNAs). siRNAs are processed, and eventually, a single strand of the siRNA is incorporated into a RISC protein complex that targets mRNA complementary to the strand included. Hence, mRNA produced by the host plant that is complementary to the strand of the RISC complex is also targeted for degradation. In this study, we explore Agrobacterium-mediated TRV-induced silencing of the phytoene desaturase (PDS) enzyme in several species of Valerianaceae that are closely related based on recent molecular phylogenetic work. Sequences were cloned and a 444 bp fragment of the gene was inserted into the TRV2 vector. Photobleaching in the leaves was used as a visual marker to signal down-regulation, or “knocking-down,” of PDS, and subsequent qRT-PCR provided empirical evidence of our ability to successfully target the host gene in planta.

Broader Impacts:

1 - St. John's University, Department of Biological Sciences, 8000 Utopia Parkway, Queens, NY, 11439, USA

Keywords:
VIGS
Valerianaceae
Agrobacterium
Fedia
TRV
Valerianella
phytoene desaturase.

Presentation Type: Poster:Posters for Topics
Session: P
Location: Grand Salon A - D/Riverside Hilton
Date: Monday, July 29th, 2013
Time: 5:30 PM
Number: PEV002
Abstract ID:631
Candidate for Awards:None