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

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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 γ 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.

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Acknowledgments

We would like to thank Dr. Konstantin Kanyuka from Biotechnology and Biological Sciences Research Council: UK (BBRSC) for his kind gift of B. distachyon seeds originally collected from Kaman. We are also grateful to International Centre for Genetic Engineering and Biotechnology (ICGEB-CRP/TUR07-03), State Planning Organization of Turkey (DPT-2004K122750) and Unit of Scientific Research Projects of METU (BAP) for their support.

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Authors and Affiliations

  1. Biochemistry Graduate Program, Middle East Technical University, Inonu Blvd., 06531, Ankara, Turkey

    Turan Demircan

  2. Department of Chemistry, Biotechnology and Biochemistry Programs, Middle East Technical University, Inonu Blvd., 06531, Ankara, Turkey

    Mahinur S. Akkaya

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  1. Turan Demircan
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  2. Mahinur S. Akkaya
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Correspondence to Mahinur S. Akkaya.

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Demircan, T., Akkaya, M.S. Virus induced gene silencing in Brachypodium distachyon, a model organism for cereals. Plant Cell Tiss Organ Cult 100, 91–96 (2010). https://doi.org/10.1007/s11240-009-9623-x

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  • DOI: https://doi.org/10.1007/s11240-009-9623-x

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