Virus induced gene silencing in Brachypodium distachyon, a model organism for cereals

Demircan T., AKKAYA M.

PLANT CELL TISSUE AND ORGAN CULTURE, vol.100, no.1, pp.91-96, 2010 (SCI-Expanded) identifier identifier

  • Publication Type: Article / Article
  • Volume: 100 Issue: 1
  • Publication Date: 2010
  • Doi Number: 10.1007/s11240-009-9623-x
  • Journal Name: PLANT CELL TISSUE AND ORGAN CULTURE
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.91-96
  • Keywords: Brachypodium distachyon, Gene silencing, GFP expression, Phytoene desaturase (PDS) gene, qRT-PCR, VIGS, REVERSE GENETICS, MESSENGER-RNA, GRASS, COLINEARITY, ARABIDOPSIS, SYSTEM, INHIBITION, RESISTANCE, ALIGNMENT, WHEAT
  • Middle East Technical University Affiliated: Yes

Abstract

Brachypodium distachyon is emerging as a model organism for crops as a better alternative to Oryzae sativa. It shares common characteristics of a model plant with its small genome, small physical plant size, a short lifecycle, and less demanding growth requirements similar to Arabidopsis thaliana. In this study, we are reporting for the first time, an implementation of virus induced gene silencing (VIGS), a powerful method allowing rapid and effective means of loss of gene function through RNA interference. To this end, Phytoene desaturase (Pds) gene, commonly preferred in gene silencing studies as a phenotypic marker, was silenced using Barley Stripe Mosaic Virus (BSMV), the most effectively used virus in monocots for VIGS., a fragment of the ORF of the B. distachyon Pds gene was cloned into the BSMV vector having the gamma genome proviral DNA. The decreased Pds gene expression was confirmed after plant infection by qRT-PCR. The effectiveness of BSMV infection was also tested with the transcripts of the vector constructed for GFP expression. We believe the demonstration of BSMV mediated VIGS will be an important step for evaluating functions of crop genes in this model organism.