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

Inactive Publication Date: 2014-06-25
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The male sterile cytoplasm used in the production of hybrid rice in my country is mainly derived from common wild rice. It can be speculated that there must be a corresponding restorer gene in wild rice (Li Shaoqing et al., 2005), but the restorer gene used in production comes from wild rice. but very few

Method used

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  • 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 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 molecular marker and special primers used for identifying a restorer line of a cytoplasmic male sterile line of common wild rice in Yuanjiang Yunnan

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses 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. The nucleotide sequence of one of the special primers used for assisting identification of the restorer line of the cytoplasmic male sterile line of the common wild rice in Yuanjiang Yunnan is shown as the sequence 1 in the sequence table, and the nucleotide sequence of the other special primer is shown as the sequence 2 in the sequence table. The molecular marker and the special primers can be used for breeding the restorer line of the cytoplasmic male sterile line of the common wild rice in Yuanjiang Yunnan, shortening the breeding period of three-line hybrid rice containing high-quality genetic resources of the common wild rice in Yuanjiang Yunnan, accelerating the breeding speed, and reducing the breeding cost. The primers and the molecular marker have advantages of simple operation, low cost, and short period, are suitable for popularization and application, and provide a rapid selection method for the restorer line having the genetic background of the common wild rice in Yuanjiang Yunnan.

Description

technical field [0001] The invention relates to a molecular marker and a special primer for identifying a restorer line of a cytoplasmic male sterile line of common wild rice Yuanjiang, Yunnan. Background technique [0002] Rice is one of the most important food crops in the world, and increasing rice yield per unit area is one of the powerful measures to ensure world food security. In the long-term evolution process of crops, because breeders select target traits according to needs and carry out directional breeding and repeated use of parents, the genetic basis of varieties is narrow. At present, more than 95% of the hybrid rice hybrids that account for more than half of the rice planting area are composed of wild-type or similar cytoplasmic male sterile lines, and long-term use of single-source cytoplasm , there may be a risk of genetic vulnerability. [0003] Wild rice is not only an important physical basis for studying the origin, classification, and evolution of ric...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13
Inventor 孙传清付强霍兴付永彩朱作峰谭禄宾刘凤霞顾凭才宏伟
Owner CHINA AGRI UNIV
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