Rice fertility recovery gene auxiliary breeding molecular marker and application thereof

A fertility restoration gene and molecular marker technology, applied in the fields of molecular biology and crop breeding, can solve the problems of long time-consuming, easy to be restricted by environmental conditions, and low selection efficiency, so as to improve work efficiency and eliminate aerosol pollution , The effect of reducing the waste of human resources

Active Publication Date: 2017-11-24
HUAZHI RICE BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] For the identification of Rf3 and Rf4 restorer genes, the marker types mainly used in existing reports are SSR and InDel markers, which have the disadvantages of low polymorphism rate and small differences in breeding
The use of EB or polyacrylamide in the detection process will easily pollute the environment and cause harm to the human body
However, the main identification method used in traditional breeding is based on phenotypic observation and identification such as seed setting rate, which takes a long time and is easily restricted by environmental conditions. There are errors in identification results and low selection efficiency.

Method used

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  • Rice fertility recovery gene auxiliary breeding molecular marker and application thereof
  • Rice fertility recovery gene auxiliary breeding molecular marker and application thereof
  • Rice fertility recovery gene auxiliary breeding molecular marker and application thereof

Examples

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Embodiment 1

[0027] Example 1 Obtaining of Rice Fertility Restoration Gene Assisted Breeding Molecular Markers

[0028] 1. Primer design

[0029] According to the published linkage markers and comparison with the reference genome, the gene Rf3 is located in the Chr.1,5598557-6278465 interval. We took the gene mapping interval as the center and expanded 50Kb to both sides, and used 3000 rice resequencing databases to extract the SNP sites in the expanded interval. Use gene donor and recipient resequencing data for analysis, SNP site mining, and at the same time, compare the cloned Rf4 gene sequence with the Nipponbare sequence, and based on the PIC value and whether the 50 bp around the SNP site contains other SNP sites, etc. Make a selection. The flanking sequences of the selected SNP sites were extracted, and primers were designed using the online primer design website BatchPrimer3 (http: / / probes.pw.usda.gov / batchprimer3 / ). There are three primers for each marker, and the 5' ends of tw...

Embodiment 2

[0051] Example 2 Verification of genetic position and phenotype of SNP markers RSRF30121 and FRF4-10-01 co-segregated with rice fertility restoration genes

[0052] 1. Marker genetic position verification

[0053] The F2 cross population (88 individual plants) of Zhonghui 8015, the donor parent of Rf3 and Rf4, and Neixiang 5A, the recipient parent, was used for marker genetic position verification. Using 15 polymorphic SNP markers in the parents and RSRF30121 and FRF4-10-01 of the present invention for genetic position analysis, the results showed that the marker RSRF30121 of the present invention was mapped to the 21.6cM position of chromosome 1. The 13 SNP markers with polymorphism in the parents and the FRF4-10-01 of the present invention were used for genetic position verification, and the marker FRF4-10-01 of the present invention was mapped to the 96.2cM position of chromosome 10.

[0054] 2. Marker phenotype verification

[0055] Using the F2 population of the donor p...

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Abstract

The invention provides a rice fertility recovery gene auxiliary breeding molecular marker, which is SNP markers RSRF30121 and FRF4-10-01 respectively co-separated from rice fertility recovery genes Rf3 and Rf4. The marker RSRF30121 is used for detecting a 5606898-th site basic group of an No.1 rice chromosome; the marker FRF4-10-01 is used for detecting a 18838172-th site basic group of an No.10 rice chromosome; primer sequences of the marker RSRF30121 developed on the basis of KASP technology are shown as SEQ ID NO:1-3. Primary sequences of the marker FRF4-10-01 developed on the basis of the KASP technology are shown as SEQ ID NO:4-6. The invention also provides application of the SNP markers RSRF30121 and FRF4-10-01 to rice fertility recovery gene auxiliary breeding. By using the marker combination, whether the rice has the fertility recovery gene or not can be fast identified; important significance is realized on promoting the application of the Rf3 and Rf4 fertility recovery genes to commercial breeding.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and crop breeding, in particular to a rice fertility restoration gene assisted breeding molecular marker and its application. Background technique [0002] Molecular marker-assisted selection breeding is the product of the combination of modern biotechnology and traditional breeding technology. It changes the breeding technology from traditional extensive and empirical to modern precision, making breeding more operable and predictable. . [0003] In the three-line hybrid rice breeding, according to the difference of the restoration relationship, there are three main types of cytoplasmic male sterility, wild defeat type (WA-CMS), Baotai type (BT-CMS) and red lotus type (HL-CMS) . Most researchers believe that wild-type cytoplasmic male sterility in rice is controlled by 2 pairs of recessive genes. Bharaj et al. (1995) believed that wild aborted male sterility is controlled by two pair...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 郑秀婷彭佩吴云天李宙炜李为国李继明肖金华
Owner HUAZHI RICE BIO TECH CO LTD
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