Dynamics of the VIGS-Mediated chimeric silencing of the Nicotiana benthamiana ChIH gene and of the tobacco mosaic virus vector

被引:40
作者
Hiriart, JB [1 ]
Aro, EM [1 ]
Lehto, K [1 ]
机构
[1] Univ Turku, Dept Plant Physiol & Mol Biol, FIN-20014 Turku, Finland
关键词
chlorophyll biosynthesis pathway; cosuppression;
D O I
10.1094/MPMI.2003.16.2.99
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The ChlH gene, encoding for the H subunit of the magnesium chelatase enzyme, was silenced in Nicotiana bentahamiana plants by virus-induced gene silencing (VIGS), using tobacco mosaic virus (TMV) expression vector. Strong silencing of the ChlH target gene was initiated only in the apical tissues, in which the endogenous transcription level of the target gene and the level of TMV vector RNA were both very high. The virus vector was also targeted by VIGS, and its suppression was correlated with the silencing of the ChlH mRNA. In the apical tissues, the suppression of both the virus vector and the ChlH mRNA led to a reduction of the silencing pressure and, consequently, to partial recovery of the new growth from the silencing. As the virus vector and the target mRNA levels increased, silencing was reestablished. The feedback regulation system, caused by the transient increase and reduction in levels of the virus vector and ChlH mRNA, led to a fluctuation of the silenced and recovered phenotypes in the plant apex. This TMV-vector mediated silencing system differed from previously analyzed VIGS systems; although the TMV vector was initially targeted by the silencing system, it was not permanently suppressed, indicating that, in this system, TMV was able to effectively escape post-transcriptional gene silencing.
引用
收藏
页码:99 / 106
页数:8
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