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Molecular marker of male nucleus-sterile gene ms39 in maize and application thereof

A molecular marker and male nucleus technology, which is applied in the fields of application, plant genetic improvement, and microbial determination/inspection, etc., can solve the problems of narrow germplasm base for breeding, heavy cross-pollination workload, and increased workload, etc., and achieve identification results. Accurate, improve breeding efficiency and reduce labor costs

Active Publication Date: 2018-05-04
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, no closely linked molecular markers have been obtained for this recessive nuclear sterility gene, which limits its further application
[0006] Breeding the two-ring line with hybrids as the basic material is the most commonly used breeding method by breeders. It has the advantages of short breeding cycle and fast breeding speed, but it will inevitably lead to narrower and narrower germplasm bases for breeding, making it difficult to breed It is a breakthrough variety, and it will face production risks due to the homogeneity of the variety
Recurrent selection is an effective method to solve the problem of narrow germplasm base, because most of the agronomic traits are controlled by quantitative genes, but the recurrent selection method has disadvantages such as long breeding cycle, heavy cross-pollination workload, and large investment. It is difficult for general breeders to engage in or persist in
The use of recessive sterility can solve the problem of heavy pollination workload, but the recessive sterility requires selfing for one generation and the homozygosity of the recessive gene in order to express genital sterility, which virtually increases the workload

Method used

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  • Molecular marker of male nucleus-sterile gene ms39 in maize and application thereof
  • Molecular marker of male nucleus-sterile gene ms39 in maize and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Gene Mapping of Maize Recessive Genic Sterility Gene ms39

[0050] (1) Construction of mapping groups

[0051] The source of maize male sterile materials used in this experiment: In 1996, the Maize Research Institute of Sichuan Agricultural University selected materials for a satellite carrying test, and obtained a male sterile mutant from it. The study found that the sterility trait is controlled by a single recessive nuclear gene , the inventor named the gene ms39.

[0052] A sterile single plant in the high-generation sister cross of the above-mentioned male sterile mutants was used as the female parent, and the inbred line Mo17 was used as the male parent to obtain F 1 Generation; F 1 F 2 generation, planting F 2 generation, to obtain trait segregation populations, and to identify F 2 The population sterile plants were isolated for generations, and the fresh leaves of the sterile plants were clipped to extract DNA for gene positioning. The specific c...

Embodiment 2

[0071] Example 2 Design and Screening of Molecular Marker Primers for Maize Recessive Genic Sterile Gene ms39

[0072] (1) Test method

[0073] (1) Primer design

[0074] According to the results of fine mapping in Example 1, the nuclear sterility gene was located between the markers M8 and M30, the physical distance was about 352Kb, and then some genes in this region were cloned and sequenced between the sterile mutant and the fertile strain. , some differential sites (SNP and indel sites) were found, and 5 pairs of primers were designed according to these differential sites (see Table 1).

[0075] Specific primers designed in Table 1

[0076] Primer name

Forward primer sequence (5'-3')

Reverse primer sequence (5'-3')

Primer 1

TCTGGCTTGGATTATTTGTCCT

CTGATTGACATGAAATCGCCTG

Primer 2

GCAGAGAACACGTCCCTT

GCTGATTGACATGAAATCGC

Primer 3

GCCAAATGTTCTTGTCACAC

ATAATCTGTGAAGGCAAGAAA

Primer 4

GTCGTCTCCAAGGTGTTCT

...

Embodiment 3

[0080] Example 3 The Molecular Marker Verification Test of the Maize Recessive Genic Sterility Gene ms39 of the Present Invention

[0081] (1) Test method

[0082] (1) Group construction

[0083] F 1 , and then self-deliver F 2 , using the inbred line Mo17 as the recurrent parent to backcross and self-cross the sterile line, and finally obtain (ms39×Mo17)BC 4 f 2 Separate groups.

[0084] (2) Identify the fertility genotype of the isolated population at the seedling stage

[0085] At the seedling stage, (ms39×Mo17)BC 4 f 2 About 5 g of fresh maize leaves were taken from a single plant of the isolated population, and genomic DNA was extracted by CTAB method. The extracted genomic DNA was used as a template and nactf was used as a primer for PCR amplification; the individual plants were marked according to the results of the amplified products. Wherein said primer nactf is:

[0086] nactf-F: 5'-TACACATGCACGCCAACATT-3' (SEQ ID NO.2);

[0087] nactf-R: 5'-GCCAACAAGTAGGACA...

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Abstract

The invention discloses a molecular marker of a male nucleus-sterile gene ms39 in maize, and belongs to the field of auxiliary selection of maize molecular marker. The molecular marker consists of a nucleotide sequence shown in SEQ ID NO.1. The invention also discloses a primer for amplifying the molecular marker. According to the molecular marker disclosed by the invention, interlocking of the molecular marker and the male nucleus-sterile gene ms39 is tight, and an identification result is accurate and reliable, so that a good tool is provided for application of the nucleus-sterile gene ms39in seed breeding; secondly, an identification method of the molecular marker disclosed by the invention is simple, and can be carried out in a seedling period; moreover, a large quantity of hybridization seed breeding requirements required by recurrent selection and the like can be met, the labor cost is reduced, the seed breeding efficiency is improved, and a great promotion effect on a seed quality basis for expanding seed breeding of the maize is achieved.

Description

technical field [0001] The invention belongs to the field of corn molecular markers, and in particular relates to a molecular marker of maize male sterility gene ms39, and also relates to the application of the molecular marker. Background technique [0002] Maize was the first plant to use cytoplasmic male sterile lines for hybrid seed production. However, in 1970, the outbreak of small class T race, which was specifically infected with T-type sterile cytoplasm, caused devastating losses to corn production in the United States. Therefore, the application of T-type male sterile cytoplasm in maize three-line hybrid seed production had to be abandoned. The use of C- and S-type sterile cytoplasms has also been limited based on the same concerns. [0003] Compared with cytoplasmic sterility genes, maize genic male sterile genes are rich in number, with a wide range of genetic background sources, simple fertility inheritance, and no problem of race-specificity of pathogenic bact...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11A01H1/02
CPCA01H1/02C12Q1/6895C12Q2600/13
Inventor 曹墨菊时子文朱永卉牛群凯李川易洪杨杨聪唐津张鹏刘孝伟董博晓魏桂夏远燕赵卓凡关淑仙牟碧涛王靖文
Owner SICHUAN AGRI UNIV
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