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

Optimisation of tobacco rattle virus-induced gene silencing in Arabidopsis

2006 ◽  
Vol 33 (4) ◽  
pp. 347 ◽  
Author(s):  
Changchun Wang ◽  
Xinzhong Cai ◽  
Xuemin Wang ◽  
Zhong Zheng
Keyword(s):  
Gene Silencing ◽  
High Throughput ◽  
Function Analysis ◽  
Tobacco Rattle Virus ◽  
Phytoene Desaturase ◽  
Growth Stages ◽  
Virus Induced Gene Silencing ◽  
Gene Functions ◽  
Genes Encoding ◽  
Gene Function Analysis

Arabidopsis thaliana (L.) Heynh. is a model plant species in which to study plant gene functions. Recently developed virus-induced gene silencing (VIGS) offers a rapid and high-throughput technique platform for gene function analysis. In this paper we report optimisation of tobacco rattle virus (TRV)-induced gene silencing in Arabidopsis. The parameters potentially affecting the efficiency of VIGS in Arabidopsis were investigated. These included the concentration and pre-incubation of Agrobacterium inocula (agro-inocula), the concentration of acetosyringone included in agro-inocula, the Agrobacterium inoculation (agro-inoculation) method, the ecotypes and the growth stages of Arabidopsis plants for agro-inoculation, and the growth temperature of agro-inoculated plants. The optimised VIGS procedure involves preparing the agro-inocula with OD600 of 2.0, pre-incubating for 2 h in infiltration buffer containing 200 μm acetosyringone, agro-inoculating by vacuum infiltration, and growth of agro-inoculated plants at 22 −24°C. Following this procedure consistent and highly efficient VIGS was achieved for the genes encoding phytoene desaturase (PDS) and actin in Arabidopsis. The silencing phenotype lasts for at least 6 weeks, and is applicable in at least seven ecotypes, including Col-0, Cvi-0, Sd, Nd-1, Ws-0, Bay-0 and Ler. TRV-induced VIGS was expressed not only in leaves, but also in stems, inflorescences and siliques. However, VIGS was not transmissible through seed to the subsequent generation. The optimised procedure of the TRV-induced gene silencing should facilitate high-throughput functional analysis of genes in Arabidopsis.

scholarly journals Virus-induced gene silencing (VIGS) in Cannabis sativa L.

Plant Methods ◽  
2019 ◽  
Vol 15 (1) ◽  
Author(s):  
Julia Schachtsiek ◽  
Tajammul Hussain ◽  
Khadija Azzouhri ◽  
Oliver Kayser ◽  
Felix Stehle
Keyword(s):  
Medicinal Plant ◽  
Gene Silencing ◽  
Cannabis Sativa ◽  
Gene Knockdown ◽  
Vector System ◽  
Carotenoid Content ◽  
Transcriptional Gene Silencing ◽  
Cotton Leaf ◽  
Virus Induced Gene Silencing ◽  
Gene Functions

Abstract Background The raised demand of cannabis as a medicinal plant in recent years led to an increased interest in understanding the biosynthetic routes of cannabis metabolites. Since there is no established protocol to generate stable gene knockouts in cannabis, the use of a virus-induced gene silencing (VIGS) method, resulting in a gene knockdown, to study gene functions is desirable. Results For this, a computational approach was employed to analyze the Cannabis sativa L. transcriptomic and genomic resources. Reporter genes expected to give rise to easily scorable phenotypes upon silencing, i.e. the phytoene desaturase (PDS) and magnesium chelatase subunit I (ChlI), were identified in C. sativa. Subsequently, the targets of specific small interfering RNAs (siRNAs) and silencing fragments were predicted and tested in a post-transcriptional gene silencing (PTGS) approach. Here we show for the first time a gene knockdown in C. sativa using the Cotton leaf crumple virus (CLCrV) in a silencing vector system. Plants transiently transformed with the Agrobacterium tumefaciens strain AGL1, carrying the VIGS-vectors, showed the desired phenotypes, spotted bleaching of the leaves. The successful knockdown of the genes was additionally validated by quantitative PCR resulting in reduced expression of transcripts from 70 to 73% for ChlI and PDS, respectively. This is accompanied with the reduction of the chlorophyll a and carotenoid content, respectively. In summary, the data clearly demonstrate the potential for functional gene studies in cannabis using the CLCrV-based vector system. Conclusions The applied VIGS-method can be used for reverse genetic studies in C. sativa to identify unknown gene functions. This will gain deeper inside into unknown biosynthetic routes and will help to close the gap between available genomic data and biochemical information of this important medicinal plant.

The Rise and Rise of Nicotiana benthamiana: A Plant for All Reasons

2018 ◽  
Vol 56 (1) ◽  
pp. 405-426 ◽  
Author(s):  
Julia Bally ◽  
Hyungtaek Jung ◽  
Cara Mortimer ◽  
Fatima Naim ◽  
Joshua G. Philips ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Protein Interactions ◽  
Nicotiana Benthamiana ◽  
Virus Induced Gene Silencing ◽  
Protein Protein Interactions ◽  
Species Origin ◽  
The Past ◽  
Gene Functions ◽  
Genetics And Genomics

A decade ago, the value of Nicotiana benthamiana as a tool for plant molecular biologists was beginning to be appreciated. Scientists were using it to study plant-microbe and protein-protein interactions, and it was the species of choice with which to activate plasmid-encoded viruses, screen for gene functions with virus-induced gene silencing (VIGS), and transiently express genes by leaf agroinfiltration. However, little information about the species’ origin, diversity, genetics, and genomics was available, and biologists were asking the question of whether N. benthamiana is a second fiddle or virtuoso. In this review, we look at the increased knowledge about the species and its applications over the past decade. Although N. benthamiana may still be the sidekick to Arabidopsis, it shines ever more brightly with realized and yet-to-be-exploited potential.

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 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
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