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Molecular marker co-separated from rice photo-thermo-sensitive genic male sterility traits and application

A technology of molecular markers and male sterility, applied in the field of plant molecular biology, can solve the problems of poor understanding of molecular basis and achieve the effect of improving the efficiency of breeding

Active Publication Date: 2020-05-05
HAINAN BOLIAN RICE GENE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many rice photothermosensitive genic male sterile genes have been mapped or cloned, the molecular basis for the regulation of rice photothermosensitive genic male sterility traits is still poorly understood

Method used

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  • Molecular marker co-separated from rice photo-thermo-sensitive genic male sterility traits and application
  • Molecular marker co-separated from rice photo-thermo-sensitive genic male sterility traits and application
  • Molecular marker co-separated from rice photo-thermo-sensitive genic male sterility traits and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Phenotype and genetic analysis of rice photothermo-sensitive genic male sterile mutant tms4430

[0043] 1) Screening of tms4430 mutants

[0044] In June 2013, 10 kg of 93-11 seeds were irradiated with cobalt 60 to obtain the M0 generation. The irradiated seeds were planted in the experimental field in Lingao County, Hainan Province, and harvested by individual plants after maturity. A total of about 6,500 M1 generation materials were obtained. In the spring of 2014, 3617 M1 generation materials with a large amount of seeds were selected and planted into plant lines, each planting 50 individual plants. Various types of mutants such as plant type, ear type, fertility, yield, etc. were selected at the tillering stage, booting stage, heading stage, flowering stage, and filling stage, and harvested and stored. One mutant was found to be male sterile in the 4430 family, and was named tms4430.

[0045] 2) Phenotypic analysis of tms4430 mutant

[0046] Under the high seaso...

Embodiment 2

[0049] Example 2 Preliminary localization of TMS4430, a photothermo-sensitive male sterility gene

[0050] Using bulked segregant analysis and recessive-class analysis to locate the TMS4430 gene. An F2 population was constructed by crossing Minghui 63 with the tms4430 mutant, and 169 sterile plants were isolated. 301 pairs of SSR markers evenly distributed on 12 chromosomes were used to screen the markers with polymorphism between 93-11 and Minghui 63, and 64 pairs of SSR markers with polymorphism were obtained. The polymorphism was low, indicating that Hui 63 and 93-11 have little difference in genome sequence. A mixed gene pool of fertile plants and sterile plants of the F2 population is constructed separately, and the fertile gene pool and the sterile gene pool are analyzed using the 64 pairs of polymorphic markers. The results showed that the SSR markers RM243 and RM023 on chromosome 1 were linked to the mutation phenotype of tms4430. The above-mentioned linkage markers we...

Embodiment 3

[0051] Example 3 Fine mapping of TMS4430, a photothermo-sensitive male sterility gene in rice

[0052] On the basis of the above coarse positioning results, a recessive-class analysis method is used for fine positioning. Using linkage markers to select tms4430 heterozygous individual plants in the F2 population developed an F3 population containing 3827 individual plants, of which 952 sterile plants were isolated. At the same time, 3 polymorphic markers were further developed between the markers RM10495 and RM10544. Among them, the primer used to detect the molecular marker Indel14270 is shown in SEQ ID NO.1-2; the primer used to detect the molecular marker Indel14340 is shown in SEQ ID NO.3-4; the primer used to detect M3193-6 is shown in SEQ ID NO. IDNO.5-7 shown.

[0053] Using these markers to detect the genotypes of the sterile plants isolated from the F3 population one by one, it was found that only the 305bp band was expanded in the Indel14270 labeled sterile plants ( ima...

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Abstract

The invention provides a molecular marker co-separated from rice photo-thermo-sensitive genic male sterility traits and an application, belonging to the technical field of gene engineering. Accordingto the invention, tms4430 is finely positioned on a rice chromosome 1 through a map-based cloning method; and the gene is located in a section of 237.39 Kb between a molecular marker RM10495 and a molecular marker RM10519, and is co-separated from markers Indel 14270, Indel 14340 and M3193-6. The tms4430 homozygous mutant shows a photo-temperature sensitive phenotype with long-day high-temperaturesterility and short-day low-temperature sterility. Through fine positioning of the tms4430 and development of a co-separation linkage marker, a novel sterile gene source and a molecular assistant selection means for breeding of a rice photo-thermo sensitive two-line sterile line are provided, and huge application potential in breeding of a two-line sterile line is achieved.

Description

Technical field [0001] The invention belongs to the field of plant molecular biology, and specifically relates to a molecular marker co-separated with rice photothermo-sensitive nuclear male sterility traits and application. Background technique [0002] Rice is the staple food for more than half of the world’s population. Increasing rice production will help humans get rid of hunger. Hybrid rice takes advantage of the heterosis phenomenon of rice, which greatly improves the yield, resistance and adaptability of rice. Since Mr. Yuan Longping proposed to use the male sterility of rice to produce hybrid rice in the 1960s, hybrid rice in China has experienced the development from three-line to two-line hybrid rice. Three-line hybrid rice uses nucleocytoplasmic sterile lines for hybrid rice breeding, while two-line hybrid rice uses light-temperature-sensitive sterile lines for hybrid rice breeding. Compared with the three-line sterile line, the two-line sterile line has a wide rang...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/11C12Q1/6895
CPCC07K14/415C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 唐杰龙湍李佳林刘昊韩晓斌曾翔吴永忠黄培劲
Owner HAINAN BOLIAN RICE GENE TECH CO LTD
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