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Outline
Developing stable, efficient, and reliable markers for purity testing of three-line hybrid seeds is crucial for current hybrid rice seed production.
Based on the specific sequences of the wild abortive-type sterility gene WA352, the major fertility-restoring gene Rf4, and the internal reference gene Actin, multiple single-gene detection markers were designed. PCR amplification was then performed by optimizing the primer mixture ratio.
A mixture of primers WA352-2, Rf4-3, and Actin-3 in a ratio of 1:11:3 enabled simultaneous detection of the genotypes of WA352, Rf4, and Actin in a single PCR reaction, allowing differentiation among wild abortive-type CMS lines, restorer lines, maintainer lines, and hybrids.
The detection method established in this study is simple to perform and yields stable and reliable results. It can not only identify genotypes in wild-abortive three-line rice but also serve as an effective tool for molecular marker-assisted breeding. By improving the accuracy of purity assessment during breeding, this method provides strong support for achieving high-yield and high-quality rice breeding objectives in China.
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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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