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Fragrance is an important trait of rice eating quality. In this study, the fragrance gene Badh2 in rice was edited using CRISPR/Cas9 technology to create a high-quality aromatic japonica rice line.
Two target sequences located in the third and fourth exons of Badh2 were selected as target sites for constructing the binary expression vector pHUE411-Badh2-gRNA. The high-quality japonica rice line Tai 20-29 was used as the transformation material.
Twenty T0 transgenic plants were obtained after transformation. Through propagation, PCR, and sequencing verification, six homozygous mutants with different base alterations in the third and fourth exons of Badh2 were identified. Except for a slightly lower seed setting rate and a higher grain number per panicle, the six mutants(bh62, bh64, bh65, bh67, bh68, and bh74) showed similar agronomic traits such as panicle length, grain length, grain width, and 1000-grain weight compared with the wild-type Tai 20-29. The mutants showed a significantly lower seed setting rate and a higher grain number per panicle compared to Tai 20-29. The 2-AP content in brown rice of mutants bh62, bh64, bh65, and the wild type was determined by GC-MS. The results showed that the 2-AP content in the mutants increased significantly, reaching 24.27, 33.89, and 38.96 mg/kg, respectively, which was much higher than that in Tai 20-29.
We successfully generated a high-quality aromatic japonica rice line by editing the Badh2 gene using CRISPR/Cas9 technology.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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