CRISPR/Cas9-Mediated Targeted Mutagenesis of BnaCOL9 Advances the Flowering Time of Brassica napus L.
Abstract
:1. Introduction
2. Results
2.1. Isolation of BnaCOL9 from Brassica napus
2.2. Knockout of BnaCOL9 Shows an Early-Flowering Phenotype
2.3. Overexpression of BnaCOL9 Delays the Flowering Time
2.4. BnaCOL9 Expression Pattern Analysis and Subcellular Localization
2.5. Transcription of Flowering-Related Genes Is Regulated by BnaCOL9
3. Discussion
4. Materials and Methods
4.1. Plant Materials
4.2. Construction of Phylogenetic Tree and Sequence Alignment
4.3. Construction of a CRISPR/Cas9 Vector
4.4. Transgenic Plant Mutation Site Identification
4.5. RNA Extraction and qRT-PCR
4.6. Subcellular Localization
4.7. GUS Assay
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Guo, J.; Zeng, L.; Chen, H.; Ma, C.; Tu, J.; Shen, J.; Wen, J.; Fu, T.; Yi, B. CRISPR/Cas9-Mediated Targeted Mutagenesis of BnaCOL9 Advances the Flowering Time of Brassica napus L. Int. J. Mol. Sci. 2022, 23, 14944. https://doi.org/10.3390/ijms232314944
Guo J, Zeng L, Chen H, Ma C, Tu J, Shen J, Wen J, Fu T, Yi B. CRISPR/Cas9-Mediated Targeted Mutagenesis of BnaCOL9 Advances the Flowering Time of Brassica napus L. International Journal of Molecular Sciences. 2022; 23(23):14944. https://doi.org/10.3390/ijms232314944
Chicago/Turabian StyleGuo, Jian, Lei Zeng, Hui Chen, Chaozhi Ma, Jinxing Tu, Jinxiong Shen, Jing Wen, Tingdong Fu, and Bin Yi. 2022. "CRISPR/Cas9-Mediated Targeted Mutagenesis of BnaCOL9 Advances the Flowering Time of Brassica napus L." International Journal of Molecular Sciences 23, no. 23: 14944. https://doi.org/10.3390/ijms232314944