Acquisition of seed dormancy breaking in rice (Oryza sativa L.) via CRISPR/Cas9-targeted mutagenesis of OsVP1 gene
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Genome editing offers great advantages in identifying gene function and generating agronomical important mutations in crops. Here, we report the development of edited lines with reduced seed dormancy by knockout viviparous-1 (OsVP1) gene known as a transcription factor that regulates key aspects of plant seed development and ABA signaling in rice. Thirty-three genetic edited lines out of 55 T0 rice plants were generated using CRISPR/Cas9 system. Sequencing analysis showed that the plants had four different mutation types at the target site of OsVP1, the mutations were found to be transmitted to the succeeding generations. Stable transmission of CRISPR/Cas9-mediated mutant lines without the transferred DNA (T-DNA) was confirmed by segregation in the T1 generation. Regarding many investigated agronomic trait, there are no significant differences between homozygous mutants and wildtype plants under field’s growth conditions. Especially in RT-PCR analysis of ABA/GA signaling genes, the expression of OsNCED2, OsGA20ox1, OsGA20ox2, OsGA20ox3 genes in homozygous mutants was increased compared to wildtype plants. Results of this study exemplified the effectiveness of CRISPR/Cas9 as a gene editing tool in broke down the seed dormancy in rice.
KeywordsCRISPR/Cas9 Gene expression Genetic transformation OsVP1 Seed dormancy
This research was supported by a Grant from the Next-Generation BioGreen 21 Program (Project no. PJ01368902), Rural Development Administration, Republic of Korea.
- Bewley JD, Bradford KJ, Hilhorst HWM, NonogakiH (2013) Seeds: physiology of development, germination and dormancy, 3rd edn. Springer, Heidelberg, pp 249–250. https://doi.org/10.1007/978-1-4614-4693-4https://doi.org/10.1007/978-1-4614-4693-4
- Gao Y, Liu J, Fan J (2011) Construction and transformation of RNAi vector of OsVP1 for a regulatory gene of pre-harvest sprouting in Oryza sativa. AGRIS 43:1321–1327Google Scholar
- Han Y, Luo D, Usman B, Nawaz G, Zhao N, Liu F, Li R (2018) Development of high yielding glutinous cytoplasmic male sterile rice (Oryza sativa L.) lines through CRISPR/Cas9 based mutagenesis of Wx and TGW6 and proteomic analysis of anther. Agronomy 8:290. https://doi.org/10.3390/agronomy8120290 CrossRefGoogle Scholar
- Koornneef M, Hanhart CJ, Hilhorst HW, Karssen CM (1989) In vivo inhibition of seed development and reserve protein accumulation in recombinants of abscisic acid biosynthesis and responsiveness mutants in Arabidopsis thaliana. Plant Physiol 90:463–469. https://doi.org/10.1104/pp.90.2.463 CrossRefPubMedPubMedCentralGoogle Scholar