Simultaneous TALEN-mediated knockout of chrysanthemum DMC1 genes confers male and female sterility

• Published in: Scientific reports Vol. 10; no. 1; p. 16165
• Main Authors: Shinoyama, Harue, Ichikawa, Hiroaki, Nishizawa-Yokoi, Ayako, Skaptsov, Mikhail, Toki, Seiichi
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 30-09-2020; Nature Publishing Group
• Subjects: 631/1647; 631/449; 631/61; Cell Cycle Proteins - genetics; Chrysanthemum - genetics; Cultivars; DNA-Binding Proteins - genetics; Gene Editing; Genome editing; Genomes; Homologous recombination; Humanities and Social Sciences; Meiosis; multidisciplinary; Plant breeding; Plant Proteins - genetics; Plants, Genetically Modified; Polyploidy; Reproduction - genetics; Science; Science (multidisciplinary); Sterility; Transcription activator-like effector nucleases; Transcription Activator-Like Effector Nucleases - genetics; Transgenic plants
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-020-72356-1

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.