Abstract
Seed weight, which is highly correlated to seed size, is a critical agronomic trait that determines the yield of Brassica napus. However, there have been limited researches on the genes involved in regulating seed size. In Arabidopsis thaliana, ENHANCER OF DA1 (EOD1), an E3 ubiquitin ligase gene, has been identified as a significant negative regulator in controlling organ size, but the function of its homologs in rapeseed remains unknown. Only two homologous of EOD1, BnaEOD1.A04 and BnaEOD1.C04, have been found in B. napus and were mutated using the CRISPR-Cas9 system. Three T-DNA-free lines, T2-157-1-C8, T2-390-2-B8, and T2-397-2-E2, were identified from the homozygous T2 mutant lines. The BnaEOD1.A04 showed a similar type of editing in these mutants, whereas the BnaEOD1.C04 in T2-397-2-E2 was only missing 26 amino acids, and the translation was not prematurely terminated, which was different from the other two mutants. In parallel, mutation of BnaEOD1s resulted in a noteworthy increase in both seed size and seed weight in the three editing lines. Additionally, there was a significant decline in the number of seeds per silique (SPS) and silique length (SL) in T2-157-1-C8 and T2-390-2-B8, but T2-397-2-E2 did not show any significant changes in the SPS and SL, possibly due to distinct types of editing in the three lines. The above results indicate the conserved function of EOD1 homologs and provides promising germplasm for breeding novel high-yield rapeseed varieties by improving seed size and thousand-seed weight.
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Data availability
The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- B. napus :
-
Brassica napus
- WT:
-
wild-type
- TSW:
-
thousand-seed weight
- T-DNA:
-
transfer DNA
- SL:
-
silique length
- SPS:
-
number of seeds per silique
- ZNF:
-
RING type zinc finger
- CDS:
-
coding sequence
- MW:
-
molecular weight
- PI:
-
isoelectric point
- Nucl:
-
nuclear
- NP:
-
number of plants used for phenotype analysis
- NS:
-
no significant difference
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This work was supported by the National Key Research and Development Program of China (2022YFD1200400) and the National Natural Science Foundation of China (32072099, 31971977)
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Material preparation, data collection, and analysis were performed by J.C. The first draft of the manuscript was written by J.G., and all authors commented on previous versions of the manuscript. D.H. participated in research design and manuscript proof. All authors read and approved the final manuscript.
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Gu, J., Chen, J., Zhao, C. et al. Mutating BnEOD1s via CRISPR-Cas9 increases the seed size and weight in Brassica napus. Mol Breeding 43, 79 (2023). https://doi.org/10.1007/s11032-023-01430-z
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DOI: https://doi.org/10.1007/s11032-023-01430-z