Identification and Characterization of Plant Genes Involved in Agrobacterium-Mediated Plant Transformation by Virus-Induced Gene Silencing
| Autor: | Ajith Anand, Zarir Vaghchhipawala, Choong-Min Ryu, Li Kang, Keri Wang, Olga del-Pozo, Gregory B. Martin, Kirankumar S. Mysore |
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| Jazyk: | angličtina |
| Rok vydání: | 2007 |
| Předmět: | |
| Zdroj: | Molecular Plant-Microbe Interactions, Vol 20, Iss 1, Pp 41-52 (2007) |
| Druh dokumentu: | article |
| ISSN: | 1943-7706 0894-0282 |
| DOI: | 10.1094/MPMI-20-0041 |
| Popis: | Genetic transformation of plant cells by Agrobacterium tu-mefaciens represents a unique case of trans-kingdom sex requiring the involvement of both bacterial virulence proteins and plant-encoded proteins. We have developed in planta and leaf-disk assays in Nicotiana benthamiana for identifying plant genes involved in Agrobacterium-mediated plant transformation using virus-induced gene silencing (VIGS) as a genomics tool. VIGS was used to validate the role of several genes that are either known or speculated to be involved in Agrobacterium-mediated plant transformation. We showed the involvement of a nodulin-like protein and an alpha-expansin protein (α-Exp) during Agrobacterium infection. Our data suggest that α-Exp is involved during early events of Agrobacterium-mediated transformation but not required for attaching A. tumefaciens. By employing the combination of the VIGS-mediated forward genetics approach and an in planta tumorigenesis assay, we identified 21 ACG (altered crown gall) genes that, when silenced, produced altered crown gall phenotypes upon infection with a tumorigenic strain of A. tumefaciens. One of the plant genes identified from the screening, Histone H3 (H3), was further characterized for its biological role in Agrobacterium-mediated plant transformation. We provide evidence for the role of H3 in transfer DNA integration. The data presented here suggest that the VIGS-based approach to identify and characterize plant genes involved in genetic transformation of plant cells by A. tumefaciens is simple, rapid, and robust and complements other currently used approaches. |
| Databáze: | Directory of Open Access Journals |
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