A molecular marker and special primers used for identifying a restorer line of a cytoplasmic male sterile line of common wild rice in Yuanjiang Yunnan
A male sterile line, wild rice technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., to achieve the effects of simple operation, reduced breeding costs, and short cycle times
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Embodiment 1
[0020] Example 1, Discovery of special primers and molecular markers
[0021] In this study, a high-generation backcross population BC was constructed with cultivated rice 93-11 as the genetic background and Yunnan Yuanjiang common wild rice as the donor. 6 f 1 , and obtained a cytoplasmic male sterile line of Oryza sativa from Yuanjiang, Yunnan, named "Yuan 9A", and the recurrent parent 93-11 was its maintainer line. This indicated that 93-11 with fertile cytoplasm could maintain the fertility of Yuan 9A, while Yuanjiang wild rice with sterile cytoplasm could restore the fertility of Yuan 9A. Yuan 9A was planted in Beijing, Hainan and Hefei, and the matching restorer line, maintainer line and sterile line confirmed that Yuan 9A was cytoplasmic male sterile.
[0022] I performed I on the pollen of Yuan 9A 2 -KI staining, it was found that the pollen of Yuan 9A was mainly lost, and could be recovered by multiple restorer lines. Testcross the sterile line with the restorer l...
Embodiment 2、489-5
[0026] Example 2. The amplification product (molecular marker) of 489-5ac in Yuanjiang wild rice and 9YJ06 is related to the fertility of the population
[0027] First according to the method of embodiment 1, to the F constructed by 9A and 9YJ06 in Yuanzhong 2 and F 3 The fertility of the population was observed for phenotype, and the sterile individual plants were selected to form a positioning population. At the same time, the genomic DNA of each sterile individual plant was extracted, and PCR amplification was carried out with primer 489-5ac in Example 1 (PCR amplification reaction and electrophoresis The conditions are the same as in Example 1), using the amplified products (molecular markers) in 9A and 9YJ06 in Yuanzhong as Markers, detect the banding type of each individual plant, analyze the genotype, and use the T-Test method to analyze the genotype and sterility Relationship. The results showed that there was a very significant correlation between primer 489-5ac and...
Embodiment 3
[0030] Example 3. Feasibility verification of screening Yuanjiang wild rice restorer genes with primer 489-5ac
[0031] Rice varieties (except Yuanzhong 9A, the following rice varieties are all from the National Germplasm Bank): Shuhui 527, Minghui 63, R6547, R507, Mianhui 501, Fuhui 838, Gui99, CDR22, Multiline 1 Hao, Mang 6, Zhongxian 2503, Digu, Wan 37B, K18B, K17B, Jin 23B, Xieqingzao, Zhenshan 97B, Yunnan Yuanjiang common wild rice and Yuan 9A. The above material was hybridized with Yuan 9A, and by F 1 Observation of pollen fertility and judgment of seed setting rate The recovery relationship between the material and Yuan 9A is shown in the table below:
[0032] Table 3. The recovery relationship between 18 rice varieties and Yuan 9A
[0033] breed name
[0034] Genomic DNA of the above-mentioned different rice varieties were extracted respectively, and PCR amplification was performed under the guidance of primer 489-5ac. The reaction system is as follows: 20...
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