Efficient transformation and artificial miRNA gene silencing inLemna minor
| Autor: | Alex Cantó-Pastor, Evan Ernst, Yiheng Yan, Jixian Zhai, Robert A. Martienssen, William Dahl, Blake C. Meyers, John Shanklin, Almudena Mollá-Morales |
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| Rok vydání: | 2014 |
| Předmět: |
DNA
Bacterial Lemna gibba Transgene Green Fluorescent Proteins Molecular Sequence Data Down-Regulation Plant Science Polymerase Chain Reaction Article Transformation Genetic Gene Expression Regulation Plant microRNA Araceae Regeneration Gene silencing Gene Silencing RNA Messenger Ecology Evolution Behavior and Systematics Genetics Lemna minor Base Sequence biology General Medicine Plants Genetically Modified biology.organism_classification Cell biology MicroRNAs Mutagenesis Insertional RNA silencing Transformation (genetics) Phenotype Magnesium chelatase |
| Zdroj: | Plant Biology. 17:59-65 |
| ISSN: | 1435-8603 |
| DOI: | 10.1111/plb.12215 |
| Popis: | Lack of genetic tools in the Lemnaceae (duckweed) has impeded full implementation of this organism as model for biological research, despite its rapid doubling time, simple architecture and unusual metabolic characteristics. Here we present technologies to facilitate high-throughput genetic studies in duckweed. We developed a fast and efficient method for producing Lemna minor stable transgenic fronds via agrobacterium-mediated transformation and regeneration from tissue culture. Additionally, we engineered an artificial microRNA (amiRNA) gene silencing system. We identified a Lemna gibba endogenous miR166 precursor and used it as a backbone to produce amiRNAs. As a proof of concept we induced the silencing of CH42, a Magnesium Chelatase subunit, using our amiRNA platform. Expression of CH42 in transgenic Lemna minor fronds was significantly reduced, which resulted in reduction of chlorophyll pigmentation. The techniques presented here will enable tackling future challenges in the biology and biotechnology of Lemnaceae. |
| Databáze: | OpenAIRE |
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