Efficient transformation and artificial miRNA gene silencing in Lemna minor

Canto-Pastor, A., Molla-Morales, A., Ernst, E., Dahl, W., Zhai, J., Yan, Y., Meyers, B. C., Shanklin, J., Martienssen, R. (January 2015) Efficient transformation and artificial miRNA gene silencing in Lemna minor. Plant Biol (Stuttg), 17 (s1). pp. 59-65. ISSN 1435-8603

URL: http://www.ncbi.nlm.nih.gov/pubmed/24989135
DOI: 10.1111/plb.12215

Abstract

Despite rapid doubling time, simple architecture and ease of metabolic labelling, a lack of genetic tools in the Lemnaceae (duckweed) has impeded the full implementation of this organism as a model for biological research. 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 L. 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.

Item Type: Paper
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene silencing
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > miRNA
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > miRNA

organism description > plant
CSHL Authors:
Communities: CSHL labs > Martienssen lab
Depositing User: Matt Covey
Date: January 2015
Date Deposited: 11 Jul 2014 16:20
Last Modified: 11 Sep 2015 14:23
PMCID: PMC4458260
Related URLs:
URI: http://repository.cshl.edu/id/eprint/30500

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