Tomato Root Transformation Followed by Inoculation with Ralstonia Solanacearum for Straightforward Genetic Analysis of Bacterial Wilt Disease
| Autor: | Alberto P. Macho, Rafael J. L. Morcillo, José Manuel García-Garrido, María Isabel Tamayo-Navarrete, Achen Zhao |
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| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
0301 basic medicine Ralstonia solanacearum biology General Immunology and Microbiology Host (biology) Agrobacterium Bacterial wilt General Chemical Engineering General Neuroscience fungi food and beverages Plant Immunity biology.organism_classification 01 natural sciences General Biochemistry Genetics and Molecular Biology Microbiology 03 medical and health sciences Transformation (genetics) 030104 developmental biology Ralstonia Pseudomonas syringae bacteria 010606 plant biology & botany |
| Zdroj: | Journal of Visualized Experiments. |
| ISSN: | 1940-087X |
| Popis: | Ralstonia solanacearum is a devastating soil borne vascular pathogen that can infect a large range of plant species, causing an important threat to agriculture. However, the Ralstonia model is considerably underexplored in comparison to other models involving bacterial plant pathogens, such as Pseudomonas syringae in Arabidopsis. Research targeted to understanding the interaction between Ralstonia and crop plants is essential to develop sustainable solutions to fight against bacterial wilt disease but is currently hindered by the lack of straightforward experimental assays to characterize the different components of the interaction in native host plants. In this scenario, we have developed a method to perform genetic analysis of Ralstonia infection of tomato, a natural host of Ralstonia. This method is based on Agrobacterium rhizogenes-mediated transformation of tomato roots, followed by Ralstonia soil-drenching inoculation of the resulting plants, containing transformed roots expressing the construct of interest. The versatility of the root transformation assay allows performing either gene overexpression or gene silencing mediated by RNAi. As a proof of concept, we used this method to show that RNAi-mediated silencing of SlCESA6 in tomato roots conferred resistance to Ralstonia. Here, we describe this method in detail, enabling genetic approaches to understand bacterial wilt disease in a relatively short time and with small requirements of equipment and plant growth space. |
| Databáze: | OpenAIRE |
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