Tobacco Rattle Virus-induced ( and () Gene Silencing in L

Overview

Journal PeerJ

Specialties Biology
Environmental Health
General Medicine

Date 2018 Mar 27

PMID 29576941

Citations 10

Authors

Hua Xu

Leifeng Xu

Panpan Yang

Yuwei Cao

Yuchao Tang

Guoren He

Suxia Yuan

Jun Ming

Affiliations

Soon will be listed here.

Abstract

Virus-induced gene silencing (VIGS) is an attractive tool for determining gene function in plants. The present study constitutes the first application of VIGS in , which has great ornamental and pharmaceutical value, using (TRV) vectors. Two marker genes, () and (), were used to test the VIGS system in . The photobleaching and yellow-leaf phenotypes of the silenced plants were shown to significantly correlate with the down-regulation of endogenous and , respectively ( ≤ 0.05). Moreover, the parameters potentially affecting the efficiency of VIGS in , including the Agrobacterium strain and the inoculation method (leaf syringe-infiltration, sprout vacuum-infiltration and seed vacuum-infiltration), were compared. The optimized VIGS parameters were the leaf syringe-infiltration method, the Agrobacterium strain GV3101 and the growth of agro-inoculated plants at 25°. With these parameters, the silencing efficiency of and could reach approximately 50% in . Additionally, the suitability of various reference genes was screened by RT-qPCR using three candidate genes, and the results demonstrated that glyceraldehyde 3-phosphate dehydrogenase () can serve as a suitable reference for assessing the gene expression levels of VIGS systems in . The proven application of VIGS in and the characterization of a suitable reference gene in the present work will expedite the functional characterization of novel genes in

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