Name: Virus-induced Gene Silencing (VIGS) in Nicotiana benthamiana and Tomato (Video) | JoVE Journal
Uploaded: 2009-06-10T00:00:00.000+00:00
Duration: 6 min 34 s
Description: Learn virus-induced gene silencing for functional genomics in Nicotiana benthamiana and tomato. This method enables transient gene knock-down using bacterial infiltration techniques.

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5 minutes

We thank Dr. Patricia Manosalva for her valuable insights on the manuscript. Funding was provided by the National Science Foundation Plant Genome Program, award number DBI-0605059.

Part 1: Plant material
N. benthamiana plants used for silencing should be around 2 ½ weeks old which is the time when the cotyledons and the first 2 - 4 true leaves have emerged. Tomato (Solanum lycopersicum) plants are used 7 - 8 days post emergence, when the true leaves have not yet appeared.
Part 2: VIGS
DAY 1
<ol>
<li>For each experiment, Agrobacterium tumefaciens harboring pTRV1, pTRV2, pTRV2-PDS and pTRV2-host target gene are grown on LB agar plates supplemented with 50 μg/mL of kanamycin and 100 μg/mL of rifampicin. The kanamycin selects for the pTRV plasmid while the rifampicin does so for the Agrobacterium. Incubate the plate at 30° C for 2 days. Silencing of PDS will cause the plants to photobleach and is used as a silencing efficiency control. Also, the genes to be downregulated must be cloned into a pTRV2 vector. There is a Gateway compatible pTRV2 vector which may facilitate the cloning and is described by Liu et al. (2002).</li>
</ol>
DAY 3
<ol>
<li>Inoculate a 2 - 3 mL liquid culture of LB with the above mentioned antibiotics for each of the strains. Incubate by shaking at 30 °C for 16 - 18 hours and 200 r.p.m.</li>
</ol>
DAY 4
<ol>
<li>Inoculate a 1 : 25 dilution of the primary culture into a secondary liquid Induction Media (IM) IM culture with kanamycin, rifampicin and 200 μM acetosyringone (Tables 1, 2 and 3). The acetosyringone is used as an inducer of the vir genes of Agrobacterium which are required for T-DNA transfer into the plant9 while the IM mimics the environment this pathogen encounters in the host apoplast. Incubate by shaking at 30 °C for 20 – 24 hours and 200 r.p.m. </li>
</ol>
DAY 5
<ol>
<li>Harvest the cells by centrifugating for 10 minutes at 3000 x g. Resuspend in the same volume that the original culture had with 10 mM MgCl2, 10 mM MES pH 5.5. Cells may be gently vortexed to resuspend them.</li>
<li>Centrifugue the cells again for 10 minutes at 3000 x g. Resuspend in half the volume of the original culture with 10 mM MgCl2, 10 mM MES pH 5.5.</li>
<li>Prepare a bacterial suspension with an O.D.600 of 0.3 for each bacterial culture. Add acetosyringone to a final concentration of 400 μM to the pTRV1 culture. </li>
<li>Mix the cultures containing the pTRV1 and the pTRV2 (or the pTRV2 containing the gene of interest) in a 1 to 1 ratio. Also include a pTRV2-PDS control. Please note that the final acetosyringone concentration is now 200 μM and that each culture is at an O.D.600 of 0.15. </li>
<li>Label the seedlings to be infiltrated with the gene to be silenced and the date of the experiment. </li>
<li>Poke a hole into each leaf to be infiltrated with a needle. Use a 1 mL needless syringe to infiltrate the bacterial suspension into the seedlings. For tomato, infiltrate both cotyledons while for N. benthamiana, infiltrate the biggest two true leaves. Five mL of each bacterial mixture should be enough to infiltrate 15 N. benthamiana and 25 tomato seedlings. Avoid cross-contamination by changing gloves between infiltrations and by not watering the plants until the next day after the inoculation.</li>
<li>The plants are kept at 20 - 22 °C in a growth chamber with a 16 hour day length and 50 % RH for at least 3 ½ weeks before they may be used for assays.</li>
</ol>
Part 3: Representative results
Figure 1 shows a representative experiment with N. benthamiana and tomato plants silenced for PDS. Plants show the characteristic photobleaching phenotype observed in plants with diminished amounts of carotenoids. For the PDS-silenced control plants, photobleaching starts to be seen as soon as 1 ½ weeks after infiltration.
<img src="/files/ftp_upload/1292/Figure%201A%201292.jpg" alt="Plant pathology experiment; Arabidopsis with leaf chlorosis, soil substrate, visual analysis of symptoms." data-altupdatedat="1787630408000" data-alt="figure 1a"><img src="/files/ftp_upload/1292/Figure%201B%201292.jpg" alt="Plant disease research, chlorosis symptom, green-yellow leaves, botany experiment, growth analysis." data-altupdatedat="1787630408000" data-alt="figure 1b">
Figure 1. Silencing of the PDS control gene causes photobleaching in N. benthamiana (A) and tomato (B) plants. Photographs were taken 3 ½ weeks after silencing.
Table 1. Preparation of Induction Medium (IM).
<table border="1" cellspacing="0" cellpadding="0">
<tbody>
<tr>
<td>400 mL</td>
<td>
distilled H2O</td>
</tr>
<tr>
<td>4.88 g</td>
<td>
MES (2-(4 morpholino)-ethane sulfonic acid)
</td>
</tr>
<tr>
<td>2.5 g</td>
<td>Glucose</td>
</tr>
<tr>
<td>0.12 g</td>
<td>
NaH2PO4
</td>
</tr>
</tbody>
</table>
Bring to a final volume of 475 mL with dH2O and adjust the pH to 5.6. Autoclave. After the medium has cooled down, add 25 mL of 20X AB salts.
Table 2. Preparation of AB salts.
<table border="1" cellspacing="0" cellpadding="0">
<tbody>
<tr>
<td>20 g</td>
<td>NH4Cl</td>
</tr>
<tr>
<td>6 g</td>
<td>MgSO4·7H2O</td>
</tr>
<tr>
<td>3 g</td>
<td>KCl</td>
</tr>
<tr>
<td>0.2 g</td>
<td>
CaCl2
</td>
</tr>
<tr>
<td>0.05 g</td>
<td>FeSO4·7H2O</td>
</tr>
</tbody>
</table>
Bring to a final volume of 1 liter with distilled water. Autoclave. Be aware that AB salts precipitate as an orange powder. Just mix them well by swirling before their use.
Table 3. Preparation of 200 mM Acetosyringone. Please note that acetosyringone should be prepared the day it will be used.
<table border="1" cellspacing="0" cellpadding="0">
<tbody>
<tr>
<td>19.6 mg</td>
<td>Acetosyringone (3’, 5’-Dimethoxy-4’-hydroxyacetophenone)</td>
</tr>
<tr>
<td>500 μL</td>
<td>
DMSO (Dimethyl sulfoxide)</td>
</tr>
</tbody>
</table>

<ol><li>Voinnet, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((RNA+silencing+as+a+plant+immune+system+against+viruses%5BTitle%5D)+AND+%22Trends+Genet%22%5BJournal%5D)">RNA silencing as a plant immune system against viruses.</a> Trends Genet. 17, 449-459 (2001).</li>
<li>Carrington, J. C., Ambros, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Role+of+microRNAs+in+plant+and+animal+development%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">Role of microRNAs in plant and animal development.</a> Science. 301, 336-338 (2003).</li>
<li>Chen, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+microRNA+as+a+translational+repressor+of+APETALA2+in+Arabidopsis+flower+development%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">A microRNA as a translational repressor of APETALA2 in Arabidopsis flower development.</a> Science. 303, 2022-2025 (2004).</li>
<li>Palatnik, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Control+of+leaf+morphogenesis+by+microRNAs%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Control of leaf morphogenesis by microRNAs.</a> Nature. 425, 257-263 (2003).</li>
<li>Brigneti, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Viral+pathogenicity+determinants+are+suppressors+of+transgene+silencing+in+Nicotiana+benthamiana%5BTitle%5D)+AND+%22EMBO+J%22%5BJournal%5D)">Viral pathogenicity determinants are suppressors of transgene silencing in Nicotiana benthamiana.</a> EMBO J. 17, 6739-6746 (1998).</li>
<li>Liu, Y., Schiff, M., Dinesh-Kumar, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Virus-induced+gene+silencing+in+tomato%5BTitle%5D)+AND+%22Plant+Journal%22%5BJournal%5D)">Virus-induced gene silencing in tomato.</a> Plant Journal. 31, 777-786 (2002).</li>
<li>MacFarlane, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Molecular+biology+of+the+tobraviruses%5BTitle%5D)+AND+%22J.+Gen.+Virol%22%5BJournal%5D)">Molecular biology of the tobraviruses.</a> J. Gen. Virol. 80, 2799-2807 (1999).</li>
<li>Senthil-Kumar, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+systematic+study+to+determine+the+extent+of+gene+silencing+in+Nicotiana+benthamiana+and+other+Solanaceae+species+when+heterologous+gene+sequences+are+used+for+virus-induced+gene+silencing%5BTitle%5D)+AND+%22New+Phytol%22%5BJournal%5D)">A systematic study to determine the extent of gene silencing in Nicotiana benthamiana and other Solanaceae species when heterologous gene sequences are used for virus-induced gene silencing.</a> New Phytol. 176, 782-791 (2007).</li>
<li>McCullen, C. A., Binns, A. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Agrobacterium+tumefaciens+and+plant+cell+interactions+and+activities+required+for+interkingdom+macromolecular+transfer%5BTitle%5D)+AND+%22Annu.+Rev.+Cell+Dev.+Biol%22%5BJournal%5D)">Agrobacterium tumefaciens and plant cell interactions and activities required for interkingdom macromolecular transfer.</a> Annu. Rev. Cell Dev. Biol. 22, 101-127 (2006).</li>
<li>Rosebrock, T. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+bacterial+E3+ubiquitin+ligase+targets+a+host+protein+kinase+to+disrupt+plant+immunity%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">A bacterial E3 ubiquitin ligase targets a host protein kinase to disrupt plant immunity.</a> Nature. 448, 370-374 (2007).</li>
<li>Miki, D., Itoh, R., Shimamoto, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((RNA+silencing+of+single+and+multiple+members+in+a+gene+family+of+rice%5BTitle%5D)+AND+%22Plant+Physiol%22%5BJournal%5D)">RNA silencing of single and multiple members in a gene family of rice.</a> Plant Physiol. 138, 1903-1913 (2005).</li>
<li>Voinnet, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Use%2C+tolerance+and+avoidance+of+amplified+RNA+silencing+by+plants%5BTitle%5D)+AND+%22Trends+Plant+Sci%22%5BJournal%5D)">Use, tolerance and avoidance of amplified RNA silencing by plants.</a> Trends Plant Sci. 13, 317-328 (2008).</li>
<li>Ekengren, S. K., Liu, Y., Schiff, M., Kumar-Dinesh, S. P., Martin, G. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Two+MAPK+cascades%2C+NPR1%2C+and+TGA+transcription+factors+play+a+role+in+Pto-mediated+disease+resistance+in+tomato%5BTitle%5D)+AND+%22Plant+J%22%5BJournal%5D)">Two MAPK cascades, NPR1, and TGA transcription factors play a role in Pto-mediated disease resistance in tomato.</a> Plant J. 36, 905-917 (2003).</li>
<li>Liu, E., Page, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Optimized+cDNA+libraries+for+virus-induced+gene+silencing+%28VIGS%29+using+tobacco+rattle+virus%5BTitle%5D)+AND+%22Plant.+Methods%22%5BJournal%5D)">Optimized cDNA libraries for virus-induced gene silencing (VIGS) using tobacco rattle virus.</a> Plant. Methods. 4, 5-5 (2008).</li>
</ol>

Virus-induced gene silencing is a method that allows rapid reverse genetic screens. It avoids the generation of T-DNA or transposon mediated gene knock-outs, which are only available in certain plants like Arabidopsis and maize. It also circumvents the time consuming process of plant transformation and allows the targeting of multiple genes at the same time, provided that they have sufficient homology10 or that different host target sequences are arranged in tandem in the silencing vector6, 11</s...

virus-induced gene silencing (vigs) in nicotiana benthamiana and tomato

Virus-induced Gene Silencing (VIGS) in Nicotiana benthamiana and Tomato

RNA interference (RNAi) is a highly specific gene-silencing phenomenon triggered by dsRNA1. This silencing mechanism uses two major classes of RNA regulators: microRNAs, which are produced from non-protein coding genes and short interfering RNAs (siRNAs). Plants use RNAi to control transposons and to exert tight control over developmental processes such as flower organ formation and leaf development2,3,4. Plants also use RNAi to defend themselves against infection by viruses. Consequently, many viruses have evolved suppressors of gene silencing to allow their successful colonization of their host5.
Virus-induced gene silencing (VIGS) is a method that takes advantage of the plant RNAi-mediated antiviral defense mechanism. In plants infected with unmodified viruses the mechanism is specifically targeted against the viral genome. However, with virus vectors carrying sequences derived from host genes, the process can be additionally targeted against the corresponding host mRNAs. VIGS has been adapted for high-throughput functional genomics in plants by using the plant pathogen Agrobacterium tumefaciens to deliver, via its Ti plasmid, a recombinant virus carrying the entire or part of the gene sequence targeted for silencing. Systemic virus spread and the endogenous plant RNAi machinery take care of the rest. dsRNAs corresponding to the target gene are produced and then cleaved by the ribonuclease Dicer into siRNAs of 21 to 24 nucleotides in length. These siRNAs ultimately guide the RNA-induced silencing complex (RISC) to degrade the target transcript2.
Different vectors have been employed in VIGS and one of the most frequently used is based on tobacco rattle virus (TRV). TRV is a bipartite virus and, as such, two different A. tumefaciens strains are used for VIGS. One carries pTRV1, which encodes the replication and movement viral functions while the other, pTRV2, harbors the coat protein and the sequence used for VIGS6,7. Inoculation of Nicotiana benthamiana and tomato seedlings with a mixture of both strains results in gene silencing. Silencing of the endogenous phytoene desaturase (PDS) gene, which causes photobleaching, is used as a control for VIGS efficiency. It should be noted, however, that silencing in tomato is usually less efficient than in N. benthamiana. RNA transcript abundance of the gene of interest should always be measured to ensure that the target gene has efficiently been down-regulated. Nevertheless, heterologous gene sequences from N. benthamiana can be used to silence their respective orthologs in tomato and vice versa8.

Description of a virus-induced gene silencing (VIGS) method for knock-down of gene expression in Nicotiana benthamiana and tomato.

RNA interference (RNAi) is a highly specific gene-silencing phenomenon triggered by dsRNA1. This silencing mechanism uses two major classes of RNA ...

The virus induced gene silencing protocol begins with growing the aphasian strains. PT RV one, PT RV two pt, RV two PT S and PT RV two target gene on LB plates. Two days later, these strains are inoculated in liquid LB and grown overnight.This culture is then used to inoculate a secondary induction medial liquid culture, which is also grown overnight. The next day, a bacterial suspension of each culture is prepared and both PT RV one and PT RV two or PT RV two target gene are mixed in a one-to-one ratio. The bacterial mixture is then infiltrated into end ana or tomato seedlings using a needleless syringe.Hi, I'm Andre Velazquez from the laboratory of Greg Martin at the Department of Plant Pathology and Plant Microbiology at Cornell University. Today we'll show you how to do virus induced gene silencing tomato and nicotiana. We use this procedure in our lab to study the interactions between a plant pathogen and its host.So let's get started. To begin this procedure for agrobacterium to fassan strains, each harboring PT RV one, PT RV two pt, RV two PDS or PT RV two host target gene plasmids are grown on LB grates supplemented with kanamycin and rifampicin. Kanamycin selects for the PTRV plasmid while rifampicin selects for the agrobacterium.Incubate the plates at 30 degrees Celsius for two days when the bacterial colonies have grown on the LB plates, use each of the strains to inoculate four, two to three milliliter cultures of liquid LB containing kanamycin and rifampicin. Incubate the cultures at 30 degrees Celsius with shaking at 200 RPM for 16 to 18 hours. On the following day, inoculate a one to 25 dilution of each primary culture into five milliliters of a secondary liquid induction media culture with can mycin rifampicin and 200 micromolar acetophenone.The IM mimics the environment that Agrobacterium encounters in the host Alast and the Acetophenone induces the bacterial VI genes which are required for TD NA transfer into the plant. Incubate the cultures at 30 degrees Celsius with shaking at 200 RPM for 20 to 24 hours. The next day harvest the cells by centrifugation for 10 minutes at 3000 gs.After removing the S supernatant, add five milliliters of 10 millimolar magnesium chloride, 10 millimolar MES pH 5.5 to the pellet and resus suspend the cells by gentle vortexing, centrifuge the cells again for 10 minutes at 3000 Gs, then resuspend the cells in half the volume of the original culture with the magnesium chloride MES solution. Prepare a bacterial suspension with an OD 600 of 0.3 for each bacterial culture. Add acetone to a final concentration of 400 micromolar to the PT RV one culture.Mix the cultures containing the PT RV one and the PT RV two or the PT RV two host target gene in a one-to-one ratio also include a PT RV two PDS control. The final acetone concentration is now 200 micromolar and each culture is at an OD 600 of 0.15. The cultures are now ready to be used for viral infiltration of the nicotiana, hamana and tomato plants and hamana plants used for silencing should be around two and a half weeks old when the coens and the first two to four true leaves have emerged.Tomato plants are used seven to eight days post emergence when the true leaves have not yet appeared. Labeled the seedlings to be infiltrated with the gene to be silenced and the date of the experiment for tomato infiltrate both olean and for Ana. Infiltrate the biggest two true leaves.Poke a hole into each leaf to be infiltrated with a needle. Use a one milliliter needleless syringe to infiltrate the bacterial suspension into the seedlings to avoid cross-contamination. Change gloves between infiltrations and do not water the plants until the day after the inoculation.After infiltration, keep the plants at 20 to 22 degrees Celsius in a growth chamber with a 16 hour day length and 50%relative humidity for at least three and a half weeks before using them for assays. This is a representative result of an experiment with ana and tomato plants three and a half weeks after being silenced for the phyto desaturated gene. Both plants show the characteristic bleaching phenotype observed in plants with diminished amounts of carotinoids.For, for PDS Silenced control plants, photobleaching starts to be seen as soon as one and a half weeks after infiltration. We've just shown you how to do virus inducing silencing in ana and tomato. In this procedure, it is important to remember to be careful to ab avoid cross contamination and to keep the plants at the proper environmental conditions that favor virus spread and silencing.So that's it. Thanks for watching and good luck with your experiments.

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We present the detailed protocol for Agrobacterium-mediated virus-induced gene silencing (VIGS) assay in cotton.  The tobacco rattle virus (TRV)-derived VIGS vectors were deployed to induce RNA silencing of cotton GrCLA1, Cloroplastos alterados 1 gene.  The albino phenotype caused by silencing GrCLA1 was observed at the seedling stage within 2 weeks after inoculation.

Agrobacterium-Mediated Virus-Induced Gene Silencing Assay In Cotton

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Optimization and Utilization of Agrobacterium-mediated Transient Protein Production in Nicotiana

Transient heterologous expression of biosynthetic enzymes in Nicotiana benthamiana leaves can divert endogenous supplies of 2,3-oxidosqualene towards the production of new high-value triterpene products. Herein is described a detailed protocol for rapid (5 days) preparative-scale production of triterpenes and analogs utilizing this powerful plant-based platform.

Transient Expression in Nicotiana Benthamiana Leaves for Triterpene Production at a Preparative Scale

Here, we present a versatile method for tomato root transformation followed by inoculation with Ralstonia solanacearum to perform straightforward genetic analysis for the study of bacterial wilt disease.

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Potato Virus X-Based microRNA Silencing (VbMS) In Potato.

An Agrobacterium-based injection (agroinjection) protocol is presented for the inoculation of foxtail mosaic virus and sugarcane mosaic virusclones into maize seedlings. Inoculation in this manner leads to viral infection, virus-induced gene silencing of marker genes, and viral overexpression of GFP.

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Learn virus-induced gene silencing for functional genomics in Nicotiana benthamiana and tomato. This method enables transient gene knock-down using bacterial infiltration techniques.

Virus-induced Gene Silencing (VIGS) in Nicotiana benthamiana and Tomato

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