Submitted to: Plant Cell Tissue And Organ Culture
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: December 31, 2012
Publication Date: April 30, 2013
Citation: Martin, R.C., Glover-Cutter, K.M., Martin, R.R., Dombrowski, J.E. 2013. Virus induced gene silencing in Lolium temulentum. Plant Cell Tissue And Organ Culture. 113:163-171. Interpretive Summary: Forage grasses are a critical component of feed used in livestock production worldwide, with many of these same species of grasses being utilized for lawns, erosion prevention, and recreation. Consequently, it is important to develop a better understanding of stress tolerance in forage and related grass species. This paper describes the use of a virus vector to facilitate the characterization of genes in forage and turf related species. Barley stripe mosaic virus (BSMV) was used as a vector to selectively reduce the expression of a marker gene. The plants infected with this vector displayed a visible phenotype approximately two weeks later, demonstrating that the BSMV vector can be used as a tool to study selected stress-related genes in the model grass species, Lolium temulentum, to identify genes that may be used to increase stress tolerance in forage and turf grasses.
Technical Abstract: Lolium temulentum L. is valuable as a model species for studying abiotic stress in closely related forage and turf grasses, many of which are polyploid outcrossing species. As with most monocot species, Agrobacterium-mediated transformation of L. temulentum is still challenging, time consuming and not very efficient. The aim of this study was to use the Barley stripe mosaic virus (BSMV) vector to develop a Virus Induced Gene Silencing (VIGS) system in L. temulentum to facilitate gene functional analysis. Plants infected with the BSMV vector containing a small region of the L. temulentum phytoene desaturase (PDS) gene, which is commonly used as a phenotypic marker in VIGS studies, displayed the characteristic albino phenotype observed in PDS silenced plants. Decreased PDS gene expression in these albino leaves was also confirmed by quantitative reverse transcriptase polymerase chain reaction. To evaluate the utility of using the VIGS vector for gene interactions in L. temulentum, a region of the translation factor eIF4A gene was incorporated into the VIGS vector. Inoculation of young seedlings with this modified vector resulted in a partial reduction in the expression of eIF4A suggesting that this vector will be a useful tool to examine gene interactions in L. temulentum.