Tang, Yang, Wang, Fei, Zhao, Jinping, Xie, Ke, Hong, Yiguo and Liu, Yule. (2010) Virus-based microRNA expression for gene functional analysis in plants. Plant Physiology, Vol.153 (No.2). pp. 632-641. ISSN 0032-0889

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Abstract

Traditional virus-induced gene silencing (VIGS) is a powerful virus-based short interfering RNA-mediated RNA silencing technique for plant functional genomics. Besides short interfering RNAs, microRNAs (miRNAs) have also been shown to regulate gene expression by RNA silencing in various organisms. However, plant virus-based miRNA silencing has not been reported. In addition, a number of plant miRNAs have been identified or predicted, while their functions are largely unknown. Thus, there is an urgent need for the development of new technologies to study miRNA function. Here, we report that a modified cabbage leaf-curl geminivirus vector can be used to express artificial and endogenous miRNAs in plants. Using this viral miRNA expression system, we demonstrate that VIGS using artificial miRNAs, dubbed as "MIR VIGS," was effective to silence the expression of endogenous genes, including PDS, Su, CLA1, and SGT1, in Nicotiana benthamiana. Silencing of SGT1 led to the loss of N-mediated resistance to Tobacco mosaic virus. Furthermore, using this viral miRNA expression system, we found that viral ectopic expression of endogenous miR156 and miR165 but not their mutants in N. benthamiana resulted in earlier abnormal developmental phenotypes, and expression of miR165 induced abnormal chlorotic spots on leaves. These results demonstrate that the cabbage leaf-curl geminivirus-based miRNA expression system can be utilized not only to specifically silence genes involved in general metabolism and defense but also to investigate the function of endogenous miRNAs in plants.

Item Type:	Journal Article
Subjects:	S Agriculture > SB Plant culture
Divisions:	Faculty of Science > Life Sciences (2010- )
Journal or Publication Title:	Plant Physiology
Publisher:	American Society of Plant Biologists
ISSN:	0032-0889
Date:	June 2010
Volume:	Vol.153
Number:	No.2
Number of Pages:	10
Page Range:	pp. 632-641
Identification Number:	10.1104/pp.110.155796
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Restricted or Subscription Access
Funder:	National Natural Science Foundation of China, National Transgenic Program of China
Grant number:	30725002, 30930060, 2008ZX08009-003, 2009ZX08001-006B
URI:	http://wrap.warwick.ac.uk/id/eprint/5758

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