| Autor: |
Sandeep Kumar, Zhan-Bin Liu, Nathalie Sanyour-Doyel, Brian Lenderts, Andrew Worden, Ajith Anand, Hyeon-Je Cho, Joy Bolar, Charlotte Harris, Lingxia Huang, Aiqiu Xing, Alexandra Richardson |
| Rok vydání: |
2021 |
| Předmět: |
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| Zdroj: |
Plant physiology. 189(2) |
| ISSN: |
1532-2548 |
| Popis: |
Gene targeting (GT) for precise gene insertion or swap into pre-defined genomic location has been a bottleneck for expedited soybean precision breeding. We report a robust selectable marker-free GT system in soybean, one of the most economically important crops. An efficient Oh H1-8 (Ochrobactrum haywardense H1-8)-mediated embryonic axis transformation method was used for the delivery of CRISPR-Cas9 components and donor template to regenerate T0 plants 6–8 weeks after transformation. This approach generated up to 3.4% targeted insertion of the donor sequence into the target locus in T0 plants, with ∼ 90% mutation rate observed at the genomic target site. The GT was demonstrated in two genomic sites using two different donor DNA templates without the need for a selectable marker within the template. High-resolution Southern-by-Sequencing analysis identified T1 plants with precise targeted insertion and without unintended plasmid DNA. Unlike previous low-frequency GT reports in soybean that involved particle bombardment–mediated delivery and extensive selection, the method described here is fast, efficient, reproducible, does not require a selectable marker within the donor DNA, and generates nonchimeric plants with heritable GT. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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