Plant-Virus Interactions
Springer US
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| Chapter title |
Replication Assay of Begomovirus in Arabidopsis Protoplasts.
|
|---|---|
| Book title |
Plant-Virus Interactions
|
| Published in |
Methods in molecular biology, January 2024
|
| DOI | 10.1007/978-1-0716-3485-1_9 |
| Pubmed ID | |
| Book ISBNs |
978-1-07-163484-4, 978-1-07-163485-1
|
| Authors |
Martins, Laura Gonçalves Costa, Fontes, Elizabeth P B, Fontes, Elizabeth P. B. |
| Abstract |
Protoplasts are isolated plant cells from which the cell walls have been removed by treatment with fungal cellulase and macerozyme enzymes, which degrade the primary components of the cell wall. The protoplasts are totipotent, sensitive, and versatile; thereby, they have been extensively used to study signal transduction pathways, cell-autonomous responses, and replication of plant viruses. This system has several advantages over the use of whole plants for viral replication, including a high percentage of infected cells and uncoupling virus movement from replication assays. Here, we describe a simple and efficient protocol for begomovirus transfection and replication in Arabidopsis protoplasts. The method consists of four steps, (i) protoplast isolation, (ii) PEG-calcium transfection of begomovirus infectious clones, (iii) elimination of input plasmid DNA by DpnI digestion, and (iv) quantification of the viral newly synthesized DNA by qPCR. The protoplasts can be transformed efficiently with begomovirus infectious clones, and virus replication can be monitored by the accumulation of nascent viral DNA in the infected protoplasts. |
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The data shown below were collected from the profile of 1 X user who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Attention Score in Context
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 21 November 2023.
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#17,038,804
of 25,038,941 outputs
Outputs from Methods in molecular biology
#5,919
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Outputs of similar age
#67,003
of 133,235 outputs
Outputs of similar age from Methods in molecular biology
#82
of 197 outputs
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