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Gene Rf5 for restoring corn C-type cytoplasmic fertility and application thereof

A maize and cytoplasmic technology, applied in the field of genes, can solve problems such as timeliness limitations, and achieve the effects of improving production efficiency, reducing breeding costs, and shortening breeding cycles

Inactive Publication Date: 2021-01-05
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The traditional CMS and restorer line transfer is mainly based on the field phenotype identification of corn plants at the pollination stage, and it takes 6-8 generations to complete the transfer, so the timeliness of selection is limited to a certain extent

Method used

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  • Gene Rf5 for restoring corn C-type cytoplasmic fertility and application thereof
  • Gene Rf5 for restoring corn C-type cytoplasmic fertility and application thereof
  • Gene Rf5 for restoring corn C-type cytoplasmic fertility and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Example 1: Obtaining the Sequence of Maize Type C Cytoplasmic Restorer Gene Rf5

[0084] The restorer gene Rf5 derived from Fengke 1 was self-crossed after 5 consecutive generations of backcrossing, and finally the Rf5 gene was introduced into the background of the inbred line Mo17 to obtain a restorer line, which was named 6233 and recorded as Cms-6233. Using the sterile line Cms-CMo17 (rf4rf4rf5rf5) and the restorer line Cms-C6233 (rf4rf4Rf5Rf5) as experimental materials, the BC1 segregation population of Rf5 was constructed. On the basis of preliminary positioning of the restorer gene Rf5 with 1,500 isolated populations, the scale of the isolated population was expanded to 32,000 strains, and molecular markers such as SSR and Indel were developed with the help of B73 genome information, and the restored gene Rf5 was finely mapped ( figure 1 ), the restorer gene Rf5 was positioned between the molecular markers chr2.09-36 and chr2.09-6+1, with a physical distance of 82...

Embodiment 2

[0097] Example 2: Rf5 corn transgenic experiment and fertility analysis of transgenic materials

[0098] In this example, the tetrad anther of the restorer line Cms-6233Rf5Rf5 was used as the material, and the full-length cDNA of the Rf5 gene was amplified by reverse transcription.

[0099] (1) Construction and transformation of Rf5 overexpression transgene vector

[0100] According to the cDNA sequence of the Rf5 gene involved primers:

[0101] p3300U-BamH1-F: 5'-cgcggatccATGCCGTCATGTGCCCGCAT-3'

[0102] p3300U-Sacl-R:5'-cgagctcTCACGGACTGGCCTCTGCAA-3'

[0103] Using the full-length cDNA of the Rf5 gene as a template, the full-length sequence of Rf5 was amplified, and the PCR product was digested with BamhI and SacI to recover the target fragment, and connected to the p3300U vector recovered by double-digestion with the same two enzymes to construct into the overexpression vector p3300U-Rf5 ( figure 2 ). The vector was introduced into maize transformation recipient inbre...

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Abstract

The invention discloses a gene Rf5 for restoring corn C-type cytoplasmic fertility and application thereof. The gene Rf5 is a separated polynucleotide and comprises a nucleotide sequence selected fromthe following nucleotide sequences: SEQ ID NOs: any one shown nucleotide sequence, a nucleotide sequence hybridized with a complementary sequence of the nucleotide sequence under a medium strict condition, preferably a high strict hybridization condition; a nucleotide sequence which is different from a protein of which the nucleotide sequence codes the same amino acid sequence but is different insequence due to the degeneracy of a genetic code; and other nucleotide sequences which are different from SEQ ID NOs 4 and 5 and are generated by selectively shearing and selecting different transcription start sites or transcription stop sites from SEQ ID NOs: 4 and 5. The nucleotide and amino acid sequences of the corn C-type cytoplasmic sterility nucleus restoring gene Rf5 are determined. By using the sequences, a corn hybrid three-line matching material can be conveniently created, the breeding period is shortened, the breeding cost is reduced, and the seed production efficiency is improved.

Description

technical field [0001] The invention relates to the field of gene technology, in particular to the gene Rf5 for restoring maize C-type cytoplasmic fertility and its application. Background technique [0002] As an important food and feed crop, corn plays an inestimable role in the development of the world economy. Corn is also the earliest crop to utilize heterosis, which plays a vital role in solving the problem of food security in our country. In the process of seed production, tassels need to be removed manually or mechanically to avoid seed mixing, which increases the production cost of hybrid seeds to a certain extent and restricts the international competitiveness of my country's seed enterprises. In 1994 and 1995, the Seed Variety Quality Monitoring Center of the Ministry of Agriculture of my country carried out random sampling tests on hybrids produced by major corn production bases in the country for two consecutive years. For seed quality standards, the number of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/46A01H1/02
CPCA01H1/02C07K14/415C12N15/8289
Inventor 汤继华薛亚东林亚楠丁冬刘宗华
Owner HENAN AGRICULTURAL UNIVERSITY