Abstract

Genome editing has become one of the key technologies for plant breeding. However, in polyploid species such as chrysanthemum, knockout of all loci of multiple genes is needed to eliminate functional redundancies. We identified six cDNAs for the CmDMC1 genes involved in meiotic homologous recombination in chrysanthemum. Since all six cDNAs harbored a homologous core region, simultaneous knockout via TALEN-mediated genome editing should be possible. We isolated the CmDMC1 loci corresponding to the six cDNAs and constructed a TALEN-expression vector bearing a CmDMC1 target site containing the homologous core region. After transforming two chrysanthemum cultivars with the TALEN-expression vector, seven lines exhibited disruption of all six CmDMC1 loci at the target site as well as stable male and female sterility at 10–30 °C. This strategy to produce completely sterile plants could be widely applicable to prevent the risk of transgene flow from transgenic plants to their wild relatives.

Details

Title
Simultaneous TALEN-mediated knockout of chrysanthemum DMC1 genes confers male and female sterility
Author
Shinoyama Harue 1 ; Ichikawa Hiroaki 2 ; Nishizawa-Yokoi Ayako 3   VIAFID ORCID Logo  ; Skaptsov Mikhail 4 ; Toki Seiichi 5   VIAFID ORCID Logo 

1  Fukui Agricultural Experiment Station, Fukui, Japan; Fukui Prefectural University, Department of Bioscience, Awara, Japan (GRID:grid.411756.0) 
2  National Agriculture and Food Research Organization (NARO), Institute of Agrobiological Sciences, Tsukuba, Japan (GRID:grid.416835.d) (ISNI:0000 0001 2222 0432) 
3  National Agriculture and Food Research Organization (NARO), Institute of Agrobiological Sciences, Tsukuba, Japan (GRID:grid.416835.d) (ISNI:0000 0001 2222 0432); Japan Science and Technology Agency (JST), Precursory Research for Embryonic Science and Technology (PRESTO), Saitama, Japan (GRID:grid.419082.6) (ISNI:0000 0004 1754 9200) 
4  Altai State University, South Siberian Botanical Garden, Barnaul, Russia (GRID:grid.77225.35) (ISNI:0000000112611077) 
5  National Agriculture and Food Research Organization (NARO), Institute of Agrobiological Sciences, Tsukuba, Japan (GRID:grid.416835.d) (ISNI:0000 0001 2222 0432); Yokohama City University, Graduate School of Nanobioscience, Yokohama, Japan (GRID:grid.268441.d) (ISNI:0000 0001 1033 6139); Yokohama City University, Kihara Institute for Biological Research, Yokohama, Japan (GRID:grid.268441.d) (ISNI:0000 0001 1033 6139) 
Publication year
2020
Publication date
2020
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1038/s41598-020-72356-1
ProQuest document ID
2449449826
Copyright
© The Author(s) 2020. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.