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

被引:31
作者
Demircan, Turan [3 ]
Akkaya, Mahinur S. [1 ,2 ]
机构
[1] Middle E Tech Univ, Dept Chem, Biotechnol Programs, TR-06531 Ankara, Turkey
[2] Middle E Tech Univ, Biochem Programs, TR-06531 Ankara, Turkey
[3] Middle E Tech Univ, Biochem Grad Program, TR-06531 Ankara, Turkey
关键词
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;
D O I
10.1007/s11240-009-9623-x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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.
引用
收藏
页码:91 / 96
页数:6
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