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Plant Biotechnol J. 2019 Apr 10. doi: 10.1111/pbi.13128. [Epub ahead of print]

Efficient genetic transformation and CRISPR/Cas9-mediated genome editing in Lemna aequinoctialis.

Liu Y1,2, Wang Y2, Xu S3, Tang X1,2, Zhao J1, Yu C2, He G2, Xu H1,2, Wang S2, Tang Y2, Fu C2, Ma Y2, Zhou G1,2.

Author information

1
College of Resources and Environment, Qingdao Agricultural University, Qingdao, China.
2
Key Laboratory of Biofuels, Qingdao Engineering Research Center of Biomass Resources and Environment, Shandong Provincial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, China.
3
Institute for Evolution and Biodiversity, University of Münster, Münster, Germany.

Abstract

The fast growth, ease of metabolic labelling and potential for feedstock and biofuels production make duckweeds not only an attractive model system for understanding plant biology, but also a potential future crop. However, current duckweed research is constrained by the lack of efficient genetic manipulation tools. Here, we report a case study on genome editing in a duckweed species, Lemna aequinoctialis, using a fast and efficient transformation and CRISPR/Cas9 tool. By optimizing currently available transformation protocols, we reduced the duration time of Agrobacterium-mediated transformation to 5-6 weeks with a success rate of over 94%. Based on the optimized transformation protocol, we generated 15 (14.3% success rate) biallelic LaPDS mutants that showed albino phenotype using a CRISPR/Cas9 system. Investigations on CRISPR/Cas9-mediated mutation spectrum among mutated L. aequinoctialis showed that most of mutations were short insertions and deletions. This study presents the first example of CRISPR/Cas9-mediated genome editing in duckweeds, which will open new research avenues in using duckweeds for both basic and applied research.

KEYWORDS:

Agrobacterium-mediated transformation; CRISPR/Cas9; duckweed; genome editing

PMID:
30972865
DOI:
10.1111/pbi.13128
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