scholarly journals Virus-induced gene silencing unravels multiple transcription factors involved in floral growth and development in Phalaenopsis orchids

2013 ◽  
Vol 64 (12) ◽  
pp. 3869-3884 ◽  
Author(s):  
Ming-Hsien Hsieh ◽  
Zhao-Jun Pan ◽  
Pei-Han Lai ◽  
Hsiang-Chia Lu ◽  
Hsin-Hung Yeh ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Gene Silencing ◽  
Growth And Development ◽  
Virus Induced Gene Silencing

scholarly journals Transcriptome analyses and virus induced gene silencing identify genes in the Rpp4-mediated Asian soybean rust resistance pathway

2013 ◽  
Vol 40 (10) ◽  
pp. 1029 ◽  
Author(s):  
Aguida M. A. P. Morales ◽  
Jamie A. O'Rourke ◽  
Martijn van de Mortel ◽  
Katherine T. Scheider ◽  
Timothy J. Bancroft ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcription Factors ◽  
Gene Silencing ◽  
Differentially Expressed ◽  
Soybean Rust ◽  
Phakopsora Pachyrhizi ◽  
R Gene ◽  
Virus Induced Gene Silencing ◽  
Asian Soybean Rust ◽  
Defence Responses

Rpp4 (Resistance to Phakopsora pachyrhizi 4) confers resistance to Phakopsora pachyrhizi Sydow, the causal agent of Asian soybean rust (ASR). By combining expression profiling and virus induced gene silencing (VIGS), we are developing a genetic framework for Rpp4-mediated resistance. We measured gene expression in mock-inoculated and P. pachyrhizi-infected leaves of resistant soybean accession PI459025B (Rpp4) and the susceptible cultivar (Williams 82) across a 12-day time course. Unexpectedly, two biphasic responses were identified. In the incompatible reaction, genes induced at 12 h after infection (hai) were not differentially expressed at 24 hai, but were induced at 72 hai. In contrast, genes repressed at 12 hai were not differentially expressed from 24 to 144 hai, but were repressed 216 hai and later. To differentiate between basal and resistance-gene (R-gene) mediated defence responses, we compared gene expression in Rpp4-silenced and empty vector-treated PI459025B plants 14 days after infection (dai) with P. pachyrhizi. This identified genes, including transcription factors, whose differential expression is dependent upon Rpp4. To identify differentially expressed genes conserved across multiple P. pachyrhizi resistance pathways, Rpp4 expression datasets were compared with microarray data previously generated for Rpp2 and Rpp3-mediated defence responses. Fourteen transcription factors common to all resistant and susceptible responses were identified, as well as fourteen transcription factors unique to R-gene-mediated resistance responses. These genes are targets for future P. pachyrhizi resistance research.

scholarly journals A High-Throughput Virus-Induced Gene-Silencing Vector for Screening Transcription Factors in Virus-Induced Plant Defense Response in Orchid

2012 ◽  
Vol 25 (6) ◽  
pp. 738-746 ◽  
Author(s):  
Hsiang-Chia Lu ◽  
Ming-Hsien Hsieh ◽  
Cheng-En Chen ◽  
Hong-Hwa Chen ◽  
Hsiang-Iu Wang ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Gene Silencing ◽  
Plant Defense ◽  
High Throughput ◽  
Viral Vector ◽  
Defense Responses ◽  
Dna Fragments ◽  
Virus Induced Gene Silencing ◽  
High Throughput Analysis ◽  
Related Plant

The large number of species and worldwide spread of species of Orchidaceae indicates their successful adaptation to environmental stresses. Thus, orchids provide rich resources to study how plants have evolved to cope with stresses. This report describes our improvement of our previously reported orchid virus-induced gene silencing vector, pCymMV-pro60, with a modified Gateway cloning system which requires only one recombination and can be inoculated by agroinfiltration. We cloned 1,700 DNA fragments, including 187 predicted transcription factors derived from an established expression sequence tag library of orchid, into pCymMV-Gateway. Phalaenopsis aphrodite was inoculated with these vectors that contained DNA fragments of the 187 predicted transcription factors. The viral vector initially triggered the expression of the salicylic acid (SA)-related plant defense responses and later induced silencing of the endogenous target transcription factor genes. By monitoring the expression of the SA-related plant defense marker PhaPR1 (homolog of PR1), we identified a gene, PhaTF15, involved in the expression of PhaPR1. Knockdown of PhaTF15 by virus-induced gene silencing and by transient delivery of double-stranded RNA (dsRNA) reduced expression of the orchid homolog of the conserved positive defense regulator NPR1, PhaNPR1. Cymbidium mosaic virus also accumulated to high levels with knockdown of PhaTF15 by transient delivery of dsRNA. We demonstrated efficient cloning and screening strategies for high-throughput analysis of orchid and identify a gene, PhaTF15, involved in regulation of SA-related plant defense.

Virus-induced Gene Silencing (VIGS) in Barley Seedling Leaves

BIO-PROTOCOL ◽  
2015 ◽  
Vol 5 (12) ◽  
Author(s):  
Lokanadha Gunupuru ◽  
Shahin Ali ◽  
Fiona Doohan ◽  
Steven Scofield
Keyword(s):  
Gene Silencing ◽  
Virus Induced Gene Silencing ◽  
Barley Seedling

scholarly journals Cucumber mosaic virus-induced gene silencing in banana

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Yuh Tzean ◽  
Ming-Chi Lee ◽  
Hsiao-Hsuan Jan ◽  
Yi-Shu Chiu ◽  
Tsui-Chin Tu ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Virus Induced Gene Silencing

scholarly journals A cassava common mosaic virus vector for virus-induced gene silencing in cassava

Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Decai Tuo ◽  
Peng Zhou ◽  
Pu Yan ◽  
Hongguang Cui ◽  
Yang Liu ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Mosaic Virus ◽  
Gene Function ◽  
Viral Vector ◽  
Function Analysis ◽  
Systemic Infection ◽  
Cdna Clones ◽  
Virus Induced Gene Silencing ◽  
Efficient Gene ◽  
Gene Function Analysis

Abstract Background Cassava is an important crop for food security and industry in the least-developed and developing countries. The completion of the cassava genome sequence and identification of large numbers of candidate genes by next-generation sequencing provide extensive resources for cassava molecular breeding and increase the need for rapid and efficient gene function analysis systems in cassava. Several plant virus-induced gene silencing (VIGS) systems have been developed as reverse genetic tools for rapid gene function analysis in cassava. However, these VIGS vectors could cause severe viral symptoms or inefficient gene silencing. Results In this study, we constructed agroinfection-compatible infectious cDNA clones of cassava common mosaic virus isolate CM (CsCMV-CM, genus Potexvirus, family Alphaflexiviridae) that causes systemic infection with mild symptoms in cassava. CsCMV-CM was then modified to a viral vector carrying the Nimble cloning frame, which facilitates the rapid and high-throughput cloning of silencing fragments into the viral genome. The CsCMV-based vector successfully silenced phytoene desaturase (PDS) and magnesium chelatase subunit I (ChlI) in different cassava varieties and Nicotiana benthamiana. The silencing of the ChlI gene could persist for more than two months. Conclusions This CsCMV-based VIGS system provides a new tool for rapid and efficient gene function studies in cassava.

scholarly journals Virus-induced gene silencing as a tool for analysis of gene functions in plants

Uirusu ◽  
2010 ◽  
Vol 60 (2) ◽  
pp. 155-162 ◽  
Author(s):  
Noriko YAMAGISHI ◽  
Nobuyuki YOSHIKAWA
Keyword(s):  
Gene Silencing ◽  
Virus Induced Gene Silencing ◽  
Gene Functions

scholarly journals Application of virus-induced gene silencing for identification of FHB resistant genes

2019 ◽  
Vol 18 (10) ◽  
pp. 2183-2192 ◽  
Author(s):  
Yan-hui FAN ◽  
Bing-qian HOU ◽  
Pei-sen SU ◽  
Hong-yan WU ◽  
Gui-ping WANG ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Virus Induced Gene Silencing ◽  
Resistant Genes

scholarly journals Simultaneous silencing of two different Arabidopsis genes with a novel virus-induced gene silencing vector

Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Kunxin Wu ◽  
Yadan Wu ◽  
Chunwei Zhang ◽  
Yan Fu ◽  
Zhixin Liu ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Double Mutant ◽  
Target Genes ◽  
Rna Binding ◽  
Protein Product ◽  
Antiviral Defense ◽  
Virus Induced Gene Silencing ◽  
Argonaute 2 ◽  
Plant Genes ◽  
Silencing Pathways

Abstract Background Virus-induced gene silencing (VIGS) is a useful tool for functional characterizations of plant genes. However, the penetrance of VIGS varies depending on the genes to be silenced, and has to be evaluated by examining the transcript levels of target genes. Results In this report, we report the development of a novel VIGS vector that permits a preliminary assessment of the silencing penetrance. This new vector is based on an attenuated variant of Turnip crinkle virus (TCV) known as CPB that can be readily used in Arabidopsis thaliana to interrogate genes of this model plant. A CPB derivative, designated CPB1B, was produced by inserting a 46 nucleotide section of the Arabidopsis PHYTOENE DESATURASE (PDS) gene into CPB, in antisense orientation. CPB1B induced robust PDS silencing, causing easily visible photobleaching in systemically infected Arabidopsis leaves. More importantly, CPB1B can accommodate additional inserts, derived from other Arabidopsis genes, causing the silencing of two or more genes simultaneously. With photobleaching as a visual marker, we adopted the CPB1B vector to validate the involvement of DICER-LIKE 4 (DCL4) in antiviral defense against TCV. We further revealed the involvement of ARGONAUTE 2 (AGO2) in PDS silencing and antiviral defense against TCV in dcl2drb4 double mutant plants. These results demonstrated that DOUBLE-STRANDED RNA-BINDING PROTEIN 4 (DRB4), whose protein product (DRB4) commonly partners with DCL4 in the antiviral silencing pathway, was dispensable for PDS silencing induced by CPB1B derivative in dcl2drb4 double mutant plants. Conclusions The CPB1B-based vector developed in this work is a valuable tool with visualizable indicator of the silencing penetrance for interrogating Arabidopsis genes, especially those involved in the RNA silencing pathways.

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